Il Dirigente della UOSD Servizio Amministrativo Ricerca

DELIBERAZIONE N. 56 DEL 19/01/2023
OGGETTO: PRESA D'ATTO STIPULA CONVENZIONE ED ACCETTAZIONE FINANZIA- MENTO DI EURO 960.000,00 DISPOSTO DAL MINISTERO DELLA SALUTE - CODICE PNRR-POC-2022-12375713 PER LO SVOLGIMENTO DEL PROGETTO DAL TITOLO:"TAR- GETING DRUG RESISTANT MELANOMA WITH MICRORNAS DELIVERED BY LIPID NANOPARTICLES (TACTIC)" RESPONSABILE XXXX. XXXXXXX XXXXXXXXX CUP H53C22001100001.
Esercizi/o 22/24 - 401030201 Centri/o di costo - - Importo presente Atto: € +960.000,00 - Importo esercizio corrente: € -	STRUTTURA PROPONENTE UOSD Servizio Amministrativo Ricerca Il Dirigente Responsabile Xxxxxxx Xxxxxx
Budget
- Assegnato: € -
- Utilizzato: € -
- Residuo: € -
Autorizzazione n°: -
Servizio Risorse Economiche: Xxxxxxxx Xxxxxxxxxxx
	Responsabile del Procedimento Xxxxxxx Xxxxxx
	L’Estensore Lucia D'Auria Proposta n° DL-30-2023
PARERE DEL DIRETTORE SANITARIO	PARERE DEL DIRETTORE AMMINISTRATIVO
Positivo	Positivo
Data 19/01/2023	Data 19/01/2023
IL DIRETTORE SANITARIO Xxxxxx Xxxxx	IL DIRETTORE AMMINISTRATIVO Xxxxx Xxxxxxxxx
Xxxxxx del Direttore Scientifico IRE Xxxxxxx Xxxxxxxxx data 18/01/2023 Positivo Parere del Direttore Scientifico ISG Xxxx Xxxxxxx data 18/01/2023 Positivo
La presente deliberazione si compone di n° 8 pagine e dei seguenti allegati che ne formano parte integrante e sostanziale: All.1 Convenzione e progetto

Il Dirigente della UOSD Servizio Amministrativo Ricerca

Visto il decreto legislativo 30.12.1992, n. 502 e successive modificazioni ed integrazioni. Visto il decreto legislativo 16.10.2003, n. 288.

Vista la legge regionale 23.01.2006, n. 2.

Visto l’Atto Aziendale adottato con deliberazione n. 153 del 19.02.2019 e approvato dalla Regione Lazio con DCA n. U00248 del 2.07.2019, modificato e integrato con deliberazioni n. 1254 del 02.12.2020, n. 46 del 21/01/2021 e n. 380 del 25.03.2021, approvate dalla Direzione Sa- lute ed Integrazione Sociosanitaria della Regione Lazio, con Determinazione n. G03488 del 30.03.2021.

Visto il Decreto del Presidente della Regione Lazio n. T00200 del 29/10/2021 avente ad oggetto: “Nomina del Direttore Generale dell’IRCCS IFO-Istituti Fisioterapici Ospitalieri.

Vista la deliberazione n.1123 del 2/11/2021 di insediamento ed assunzione in carica del Direttore Generale degli Istituti Fisioterapici Ospitalieri di Roma Dott. ssa Xxxxxx Xxxxxxxx.

Viste le deliberazioni n. 212 del 16/03/2022 e n. 154 del 28/02/2022 con le quali sono stati nomi- nati rispettivamente la Dott. ssa Xxxxx Xxxxxxxxx quale Direttore Amministrativo ed il Xxxx. Xxxxxx Xxxxx quale Direttore Sanitario degli Istituti Fisioterapici Ospitalieri.

Visto il D.M. del Ministero della Salute del 8 maggio 2020 di conferma del riconoscimen to del ca- rattere scientifico dell’IRCCS di diritto pubblico a Istituti Fisioterapici Ospitalieri (IFO) re- lativamente alla disciplina di “oncologia” per l’Istituto Nazionale Tumori Regina Xxxxx (IRE) e alla disciplina di “dermatologia” per l’Istituto San Gallicano;

Premesso che l’art. 7 del decreto legislativo 16 ottobre 2003 n. 288, contempla le diverse tipologie di rica- vi degli IRCCS;

che l’art. 8 del D.Lgs. n. 288/2003 prevede la possibilità per gli IRCCS di stipulare accordi e convenzioni, costituire e/o partecipare a consorzi e attuare misure di collegamento e sinergia con altre strutture di ricerca e assistenza sanitaria, pubbliche e private, nonché con le Universi- tà, per la realizzazione di comuni progetti di ricerca;

che il vigente Regolamento di Organizzazione e Funzionamento degli IFO definisce, fra l’altro, come missione degli IRCCS la cooperazione con altri enti pubblici di ricerca e con altre organizzazioni che operano negli specifici campi, in una logica di completamento di ruoli e di continuità assistenziale;

che con deliberazione n. 146 del 24/02/2022 il Direttore Generale degli IRCCS IFO, in ordine alla stipula degli accordi di riservatezza con i partner degli Istituti, degli MTA e degli MTDA relativi alle sperimentazioni cliniche, agli studi osservazionali, ai brevetti e comunque ai pro- getti di ricerca ha esteso delega ai Direttori Scientifici IRE e ISG per le attività afferenti i ri - spettivi Istituti.

Rilevato che il decreto del Ministro della salute 1° aprile 2022 che nella relativa tabella ha previsto ai punti

2.1.1 - proof of concept, 2.1.2 – tumori e malattie rare e 2.1.3 – malattie altamente invalidanti, la ripartizione degli interventi di investimento

che è stato previsto un bando per la presentazione e selezione di progetti di ricerca da finanziare nell’ambito del PNRR, pubblicato sul sito web del Ministero della Salute il 20 aprile 2022 e sulla gazzetta ufficiale della Repubblica italiana, sulle seguenti tematiche: Proof of concept (PoC);

che con il decreto direttoriale n. 27 del 2 novembre 2022, registrato con Visto n. 1054 dall’Ufficio centrale di bilancio in data 18 novembre 2022, con il quale è stata approvata la graduatoria dei progetti di ricerca PNRR ed i relativi Principal Investigator;

Considerato che con il messaggio trasmesso per il tramite della piattaforma WorkFlow della ricerca in data 13 dicembre 2022 è stato comunicato che la valutazione della proposta progettuale ha avuto esito po- sitivo e che, pertanto, la stessa è stata ammessa a finanziamento;

che al progetto con codice PNRR-POC-2022-12375713 dal titolo TArgeting drug resistant mela- noma with miCroRNAs delivered by Lipid NanoparTICles (TACTIC) Destinatario Istituzionale - Istituti Fisioterapici Ospitalieri - Istituto Nazionale Tumori Regina Xxxxx – di cui il Xxxx. Xxxxxxx Xxxxxxxxx è Principal Investigator e utilmente collocato nella specifica graduatoria, è stato attribuito un finanziamento complessivo di euro 960.000,00 (novecentosessantaamila/00);

che l’importo concesso a finanziamento è pari a circa l’80% del totale del budget presentato, come si evince dal budget allegato alla convenzione (rif. pag. 70 All.1)

Visto che tra il Ministero della Salute e l’IFO IRCCS IRE è stata stipulata la Convenzione facente rife- rimento al progetto POC con codice PNRR-POC-2022- 12375713, (Allegato 1), ai fini dello svol- gimento del progetto di ricerca e per disciplinare i rapporti tra le parti e che, la stessa allegata in copia conforme all’originale, rappresenta parte integrante e sostanziale del presente atto;

Rilevato che gli Istituti Fisioterapici Ospitalieri – Istituto Nazionale Tumori Xxxxxx Xxxxx hanno presentato il progetto, oggetto del presente atto, in qualità di Destinatario Istituzionale e tramite il Principal Investigator, Dr. Xxxxxxx Xxxxxxxxx, svolgerà lo stesso secondo quanto riportato nel piano esecutivo approvato dal Ministero, agli atti della scrivente;

che la convenzione ha la durata di 24 mesi prorogabile eventualmente di ulteriori 6 mesi come pre- visto dall’ articolo 11.

che l’attività di ricerca, da svolgersi nell’arco temporale della vigenza della convenzione, deve avere inizio improrogabilmente entro e non oltre il 20 maggio 2023, comunicando la data effettiva di avvio con nota sottoscritta digitalmente dal proprio rappresentante legale e dal Principal investi- gator della ricerca che deve essere trasmessa almeno 30 giorni prima dell’inizio effettivo, correlata di documentazione idonea;

Atteso che le procedure di erogazione dei fondi su richiesta del Soggetto attuatore/beneficiario a titolo di anticipazione e a titolo di rimborso all’Unità di missione del Ministero della salute seguono le spe- cifiche modalità in conformità con quanto indicato nell’Avviso e di seguito riportate:

- massimo 40% al momento della comunicazione, da parte del Soggetto beneficiario, dell’inizio dell’attività di ricerca, a titolo di anticipazione.

- quota a rimborso per un ulteriore per massimo un complessivo pari all’80% dopo l’invio, al 12° mese dall’inizio delle attività progettuali, da parte del Soggetto attuatore/beneficiario della relazione scientifica intermedia e dopo la sua approvazione, sulla base della presentazione delle richieste di pagamento a titolo di rimborso per le spese effettivamente sostenute dal Soggetto beneficiario.

- quota a rimborso residuale a saldo, a conclusione della ricerca, dopo l’invio da parte del Soggetto attuatore/beneficiario della relazione scientifica finale e della rendicontazione economica, sulla base della presentazione della richiesta di pagamento finale attestante la conclusione del progetto.

Visto che alla realizzazione del progetto parteciperanno gli IFO IRCCS IRE e altre n° 3 Unità Operati- ve: Università La Sapienza, Università Xxxxxxxx XX e Istituto Nazionale Tumori IRCCS Fondazio- ne Xxxxxxx, la cui distribuzione del finanziamento è la seguente:

Costs	UO1 IFO		OU2 S +apienza	UO3 sità II	Univer- Xxxxxxxx	UO4 IRCCS Xxxxxxx	Totale		%
1 Staff Salary	not permitted		not permit- ted	not permitted		not permitted	€	-	0,00%
2 Researchers' Contracts	00 €	50.000,	50.0 00,00 €	00 €	50.000,	100.00 0,00 €	€	250.000,00	26,04%
3a.1 Equipment (Leasing - Rent)	- €		- €	- €		- €	€	-	0,00%
3a.2 Equipment (buying)	- €		- €	- €		- €	€	-	0,00%
3b Supplies	00 €	146.500,	162.2 00,00 €	00 €	157.550,	72.55 0,00 €	€	538.800,00	56,13%
3c Model Costs	00 €	20.000,	- €	- €		35.00 0,00 €	€	55.000,00	5,73%
4 Subcontracts	- €		- €	- €		- €	€	-	0,00%
5 Patient Costs	- €		- €	- €		- €	€	-	0,00%
6 IT Services and Data Bases	- €		- €	- €		- €	€	-	0,00%
7 Travels	00 €	4.000,	4.0 00,00 €	00 €	4.000,	4.00 0,00 €	€	16.000,00	1,67%
8 Publication Costs	00 €	5.000,	5.0 00,00 €	00 €	5.000,	5.00 0,00 €	€	20.000,00	2,08%
9 Dissemination	2.000,		2.0	2.000,		2.00	8.000,00		0,83%

	00 €	00,00 €	00 €	0,00 €	€
10 Overheads	17.500, 00 €	16.8 00,00 €	16.450, 00 €	16.45 0,00 €	67.200,00 €	7,00%
11 Coordination Costs	5.000, 00 €	not permit- ted	not permitted	not permitted	5.000,00 €	0,52%
Totale	250.000, 00 €	240.0 00,00 €	235.000, 00 €	235.00 0,00 €	960.000,00 €	100%

Visto che il codice CUP del progetto, oggetto del presente atto, è H53C22001100001;

Preso atto della nota folium n. 16838 del 15 dicembre 2022 con cui la Direzione Scientifica IRE richiedeva di provvedere alla relativa procedura deliberativa della convenzio- ne allegata;

Ritenuto opportuno prendere atto della Convenzione relativa al progetto con codice PNRR-POC- 2022-12375713 dal titolo TArgeting drug resistant melanoma with miCroRNAs delivered by Lipid NanoparTICles (TACTIC) Destinatario Istituzionale - Istitu- ti Fisioterapici Ospitalieri - Istituto Nazionale Tumori Regina Elena – di cui il Xxxx. Xxxxxxx Xxxxxxxxx è Principal Investigator e unitamente accettare il finan- ziamento complessivo di euro 960.000,00 (novecentosessantaamila/00);

Attestato che il presente provvedimento, a seguito dell’istruttoria effettuata, nella forma e nella sostanza è totalmente legittimo e utile per il servizio pubblico, ai sensi dell’art. 1 della legge 20 del 14/01/1994 e successive modifiche, nonché alla stregua dei criteri di economicità e di efficacia di cui all’art. 1, primo comma, della legge 241 del 7/08/1990, come modificata dalla legge 15 del 11/02/2005;

Propone

Per i motivi di cui in narrativa che si intendono integralmente confermati di:

• prendere atto della Convenzione relativa al progetto con codice PNRR-POC-2022-12375713 dal titolo TArgeting drug resistant melanoma with miCroRNAs delivered by Lipid NanoparTICles (TACTIC) di cui il Xxxx. Xxxxxxx Xxxxxxxxx è Principal Investigator

• accettare il finanziamento complessivo di euro 960.000,00 (novecentosessantaamila/00) Desti- xxxxxxx Istituzionale - Istituti Fisioterapici Ospitalieri - Istituto Nazionale Tumori Regina Elena;

• dare mandato al Servizio Risorse Economiche di iscrivere al piano dei conti n. 401030201 la somma di euro 960.000,00 (novecentosessantaamila/00).

La UOSD Servizio Amministrativo per la Ricerca curerà tutti gli adempimenti per l’esecuzione del- la presente deliberazione.

Il Dirigente della UOSD Servizio Amministrativo Ricerca

Xxxxxxx Xxxxxx

Il Direttore Generale

Visto il Decreto Legislativo 30.12.1992, n. 502 e successive modificazioni ed integrazioni; Vista la Legge Regionale 23.01.2006, n. 2;

Visto l’Atto Aziendale adottato con deliberazione n. 153 del 19.02.2019 e approvato dalla Regione Lazio con DCA n. U00248 del 2.07.2019, modificato e integrato con delibe- razioni n. 1254 del 02.12.2020, n. 46 del 21/01/2021 e n. 380 del 25.03.2021, appro- vate dalla Direzione Salute ed Integrazione Sociosanitaria della Regione Lazio, con Determinazione n. G03488 del 30.03.2021;

In virtù dei poteri conferitigli con Decreto del Presidente della Regione Lazio n. T00200 del 29.10.2021.

Preso atto che il Dirigente proponente il presente provvedimento, sottoscrivendolo, attesta che lo stesso a seguito dell’istruttoria effettuata, nella forma e nella sostanza è totalmente legittimo e utile per il servizio pubblico, ai sensi dell’art. 1 della legge 20/94 e s.m.i., nonché alla stregua dei criteri di economicità e di efficacia di cui all’art. 1, primo comma, della legge 241/90, come modificata dalla legge 15/2005.

Visto il parere favorevole del Direttore Amministrativo e del Direttore Sanitario Aziendale; ritenuto di dover procedere;

Delibera

di approvare la proposta così formulata concernente “PRESA D'ATTO STIPULA CONVENZIONE ED AC- CETTAZIONE FINANZIAMENTO DI EURO 960.000,00 DISPOSTO DAL MINISTERO DELLA SALUTE - CODICE PNRR-POC-2022-12375713 PER LO SVOLGIMENTO DEL PROGETTO DAL TITOLO:"TARGETING DRUG RESI- STANT MELANOMA WITH MICRORNAS DELIVERED BY LIPID NANOPARTICLES (TACTIC)" RESPONSABILE

XXXX. XXXXXXX XXXXXXXXX CUP H53C22001100001.” e di renderla disposta.

Il Direttore Generale Dr.ssa Xxxxxx Xxxxxxxx

Documento firmato digitalmente ai sensi del D.Lgs 82/2005 s.m.i. e norme collegate

PIANO NAZ IONALE DI RIPRESA E RESILIENZ A (PNRR) MISSIONE 6 - COMPONENTE 2

INVESTIMENTO 2.1 VALORIZ Z AZ IONE E POTENZ IAMENTO DELLA RICERCA

BIOMEDICA DEL SSN

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s a l u t e , i l So g g e t t o

Ias tt ti

ut au tt oi

r f e i /bs ei

no et fe ir ca ip ai

rc ii

o o s pei

it la l i e

Pr i n c i p a l In v e s t i gGEa NtNoARr O dCIeLIl BlERaTOr, i pc ee rr cl aa r e g o l a m e n t a z

s v o l g i m e n t oPrdoe ol f por f o cg oe nt cct oeo np tc o(Pdo iC)c XxXXx Xx-XxXxX-x00x00x-00x000000, d a l

t i t ToArl

(TACTIC);

go e t i n g d r u g r e s i s t a n t m e l a n o m a w i t h m i Cr o RNAs d

Pr e m e s s o c h e

VISTA l a l e g g e 7 a g o s t o 1990, n . 241 “ Nu o v e n o r m e i n m a t e r i a d i d i r i t t o d i a c c e s s o a i d o c u m e n t i a m m i n i s t r a t i v i ” e s .m VISTA l a l e g g e 14 g e n n a i o 1994 n . 20 “ Di s p o s i z i o n i i n m a t e r i a d e i Co n t i ” e s .m .i .;

VISTO l ’ a r t i c o l o 12 b i s , c o m m a 3, d e l d e c r e t o l e g i s l a t i v o 3 VISTO i l d e c r e t o d e l Pr e s i d e n t e d e l Co n s i g l i o d e i Mi n i s t d i o r g a n i z z a z i o n e d e l Mi n i s t e r o d e l l a s a l u t e e , i n p a r VISTO i l d e c r e t o d e l Pr e s i d e n t e d e l l a Re p u b b l i c a 28 m a r z o d e g l i o r g a n i c o l l e g i a l i e d e g l i a l t r i o r g a n i s m i o p e r

a r t t . 3 e 4 c h e p r e v e d o n o l a c o m p o s i z i o n e d e l Co m i t a t o t e VISTO i l d e c r e t o d e l Mi n i s t r o d e l l a s a l u t e 8 a g o s t o 0000, x x x x x Xx n i s t e r o d e l l a s a l u t e i n d a t a 13 a g o s t o 0000, x x x t o n . 9 r i p a r t i z i o n e d e i c o m p o n e n t i t r a l e s e z i o n i d e l Co m i t a VISTO i l d e c r e t o d e l Mi n i s t r o d e l l a s a l u t e 15 d i c e m b r e 0000 x x x x x x x x Xx n i s t e r o d e l l a s a l u t e i n d a t a 7 g e n n a i o 2022, v t e c n i c o s a n i t a r i o , a v e n t e u n a d u r a t a d i t r e a n n i d a l l a VISTO i l Re g o l a m e n t o (UE) 2021/241 d e l Pa r l a m e n t o e u r o p e o e d e l

i s t i t u i s c e i l d i s p o s i t i v o p e r l a r i p r e s a e l a r e s i l i e VISTO i l Pi a n o Na z i o n a l e d i Ri p r e s a e Re s i l i e n z a (PNRR) v a l Co n s i g l i o ECOFIN d e l 13 l u g l i o 2021, n o t i f i c a t a a l l ’ It a l i a d n o t a LT161/21, d e l 14 l u g l i o 2021, e d i n p a r t i c o l a r e l a Mi s s i o “ Va l o r i z z a z i o n e e p o t e n z i a m e n t o d e l l a r i c e r c a b i o m e

d e l l a r i c e r c a b i o m e d i c a t r a m i t e d u e l i n e e d i i n t e r v e (Po C), s o s t e n e n d o l o s v i l u p p o d i t e c n o l o g i e c o n u n b a s s o i l t r a s f e r i m e n t o d i t e c n o l o g i e v e r s o l 'i n d u s t r i a ; b ) i l c a m p o d e l l e m a l a t t i e r a r e e d e i t u m o r i r a r i e d i a l t r e VISTO i l Re g o l a m e n t o (UE) 2018/1046 d e l 18 l u g l i o 2018, c h e s t a b i l i s a l b i l a n c i o g e n e r a l e d e l l ’ Un i o n e , c h e m o d i f i c a i Re g 1303/2013, n . 1304/2013, n . 1309/2013, n . 1316/2013, n . 223/2014, n . 283/2014 e l a d e c i s i

541/2014/UE e a b r o g a i l r e g o l a m e n t o (UE, Eu r a t o m ) n . 966/2012;

VISTO i l d e c r e t o l e g g e d e l 31 m a g g i o 2021, n . 77, c o n v e r t i t o c o 2021, n . 108 « Go v e r n a n c e d e l Pi a n o n a z i o n a l e d i r i p r e s a e r e d e l l e s t r u t t u r e a m m i n i s t r a t i v e e d i a c c e l e r a z i o n e e VISTO i l d e c r e t o d e l Pr e s i d e n t e d e l Co n s i g l i o d e i m i n i s t a m m i n i s t r a z i o n i c e n t r a l i t i t o l a r i d i i n t e r v e n t i p r

c i t a t o d e c r e t o l e g g e 31 m a g g i o 2021, n . 77, c o n v e r t i t o , c o n m 108;

VISTO i l d e c r e t o d e l Mi n i s t r o d e l l ’ e c o n o m i a e d e l l e f i n d e l l e r i s o r s e i n f a v o r e d i c i a s c u n a Am m i n i s t r a z i o n e m i l e s t o n e e t a r g e t ;

VISTO i l d e c r e t o d e l Mi n i s t r o d e l l a s a l u t e , d i c o n c e r t o c s e t t e m b r e 2021, d i i s t i t u z i o n e d e l l ’ Un i t à d i Mi s s i o n e PNRR, a i s e n s i d e l l ’ a r t i c o l o 8 d e l c i t a t o d e c r e t o l e g g e n VISTO l ’ a t t o d i i n d i r i z z o d e l Mi n i s t r o d e l 12 o t t o b r e 2021 So g g e t t i At t u a t o r i n e l l ’ a m b i t o d e g l i i n t e r v e n t i e s u r i p r e s a e r e s i l i e n z a (PNRR) a t i t o l a r i t à d e l Mi n i s t e r o d VISTO i l d e c r e t o l e g g e 6 n o v e m b r e 2021, n . 152 “ Di s p o s i z i o n i n a z i o n a l e d i r i p r e s a e r e s i l i e n z a (PNRR) e p e r l a p r e v e n z VISTA l a l e g g e 16 g e n n a i o 2003, n . 3 “ Di s p o s i z i o n i o r d i n a a m m i n i s t r a z i o n e ” e , i n p a r t i c o l a r e , l ’ a r t i c o l o 11, c o m m a n c h e d i n a t u r a r e g o l a m e n t a r e a d o t t a t i d a l l e Am m i n i s l e g i s l a t i v o 30 m a r z o 2001, n . 165, c h e d i s p o n g o n o i l f i n a n z i a d i p r o g e t t i d i i n v e s t i m e n t o p u b b l i c o , s o n o n u l l i i n a s

c o s t i t u i s c o n o e l e m e n t o e s s e n z i a l e d e l l 'a t t o s t e s s o ” VISTA l a d e l i b e r a d e l CIPE n . 63 d e l 00 x x x x x x r e 2020 c h e i n t r o d u c d e l CUP;

VISTO l ’ a r t i c o l o 1, c o m m a 1042, d e l l a l e g g e 30 d i c e m b r e 2020, n . 1 d e c r e t i d e l Mi n i s t r o d e l l ’ e c o n o m i a e d e l l e f i n a n z e s o l a g e s t i o n e d e l l e r i s o r s e d i c u i a i c o m m i d a 1037 a 1050, n o g e s t i o n e d e l Fo n d o d i c u i a l c o m m a 1037;

VISTO l ’ a r t i c o l o 1, c o m m a 1043, s e c o n d o p e r i o d o , d e l l a l e g g q u a l e a l f i n e d i s u p p o r t a r e l e a t t i v i t à d i g e s t i o n e , d i c o m p o n e n t i d e l Ne x t Ge n e r a t i o n EU, i l Mi n i s t e r o d e l l 'e Ra g i o n e r i a g e n e r a l e d e l l o St a t o s v i l u p p a e r e n d e d i s p VISTO l ’ a r t i c o l o 17 d e l Re g o l a m e n t o (UE) 2020/852 c h e d e f i n i s c p r i n c i p i o d i n o n a r r e c a r e u n d a n n o s i g n i f i c a t i v o (DNSH d e l l a Co m m i s s i o n e UE 2021/C 58/01“ Or i e n t a m e n t i t e c n i c i s u l l u n d a n n o s i g n i f i c a t i v o » a n o r m a d e l r e g o l a m e n t o s u l d VISTI i p r i n c i p i t r a s v e r s a l i p r e v i s t i d a l PNRR, q u a l i , t r a c l i m a t i c o e d i g i t a l e (c .d . t a g g i n g ), i l p r i n c i p i o d i p a r i d e i g i o v a n i ;

VISTI g l i o b b l i g h i d i a s s i c u r a r e i l c o n s e g u i m e n t o d i t a r n e l PNRR;

VISTO i l Re g o l a m e n t o d e l e g a t o (UE) 2021/2106 d e l l a Co m m i s s i o n e i l r e g o l a m e n t o (UE) 2021/241 d e l Pa r l a m e n t o e u r o p e o e d e l Co n r i p r e s a e l a r e s i l i e n z a , s t a b i l e n d o g l i i n d i c a t o r i c o d e l l a r i p r e s a e d e l l a r e s i l i e n z a , c h e p r e v e d e , i n p a r t i g l i i n d i c a t o r i c o m u n i , s i a a g g i o r n a t o i n m o d o c o e r e n t

r i f e r i s c o n o a l l a Co m m i s s i o n e d u e v o l t e l ’ a n n o n e l l ’ a n e l l a r e a l i z z a z i o n e d e i p i a n i p e r l a r i p r e s a e l a r e s i

c o m u n i .”

VISTE l e “ Li n e e Gu i d a p e r l o s v o l g i m e n t o d e l l e a t t i v i t à c

d a l Se r v i z i o Ce n t r a l e p e r i l PNRR, p r e s s o i l Mi n i s t e r o Di p a r t i m e n t o Ra g i o n e r i a g e n e r a l e d e l l o St a t o (RGS), c h

i n f o r mRa et Gi”iv Sso v “i l u p p a t o d a l Mi n i s t e r o d e l l ’ e c o n o m i a e Ra g i o n e r i a Ge n e r a l e d e l l o St a t o i n a t t u a z i o n e d e l l ’ a r n . 178;

VISTO i l d o c u m e n t o “ Si s t e m a d i Ge s t i o n e e Co n t r o l l o (Si .G a d o t t a t o c o n De c r e t o d e l 29 l u g l i o 2022;

VISTE l e “ Li n e e Gu i d a p e r l o s v o l g i m e n t o d e l l e a t t i v i t à d d i c o m p e t e n z a d e l l e Am m i n i s t r a z i o n i c e n t r a l i e d e i So p e r i l PNRR, p r e s s o i l Mi n i s t e r o d e l l ’ e c o n o m i a e d e l l e f i d e l l o St a t o (RGS), c h e c o n t e n g o n o i n d i c a z i o n i p r o c e d u r a c o n t r o l l o e r e n d i c o n t a z i o n e d e l l e s p e s e e d i Mi l e s t o n n o r m a t i v a c o m u n i t a r i a e n a z i o n a l e a p p l i c a b i l e a l PNR 77 d e l 31 m a g g i o 2021, c o m e m o d i f i c a t o d a l l a l e g g e d i c o n v e r VISTO i l d e c r e t o d e l Pr e s i d e n t e d e l Co n s i g l i o d e i Mi n i s s t r u m e n t i p e r i l c o n f e r i m e n t o d e i d a t i ” ;

VISTA l a Ci r c o l a r e MEF-RGS d e l 14 o t t o b r e 2021, n . 21 “ Pi a n o Na z i (PNRR) - Tr a s m i s s i o n e d e l l e Is t r u z i o n i Te c n i c h e p e r l a s e l VISTO i l De c r e t o i n t e r m i n i s t e r i a l e d e l 7 d i c e m b r e 2021 p e r l a p a r i o p p o r t u n i t à d i g e n e r e e g e n e r a z i o n a l i , n o n c h é n e i c o n t r a t t i p u b b l i c i f i n a n z i a t i c o n l e r i s o r s e d e l VISTA l a Ci r c o l a r e MEF-RGS d e l 30 d i c e m b r e 2021, n . 00, x x x x x x e “ Gu

p r i n c i p i o d i n o n a r r e c a r e d a n n o s i g n i f i c a t i v o a l l ’ a m VISTA l a Ci r c o l a r e MEF-RGS d e l 31 d i c e m b r e 2021, n . 33 “ Pi a n o Na z i (PNRR) – No t a d i c h i a r i m e n t o s u l l a Ci r c o l a r e d e l 14 o t t o b r Te c n i c h e p e r l a s e l e z i o n e d e i p r o g e t t i PNRR – Ad d i z i o n a

a s s e n z a d e l c .d . d o p p i o f i n a n z i a m e n t o ”

VISTA l a Ci r c o l a r e MEF-RGS d e l 21 g i u g n o 2022, n . 27 “ Mo n i t o r a g g i VISTA l a Ci r c o l a r e MEF-RGS d e l l ’ 11 a g o s t o 2022, n . 30 s u l l e p r o c e d e l l e m i s u r e PNRR;

VISTA l a Co m u n i c a z i o n e d e l l a Co m m i s s i o n e 2014/C 198/01 “ Di s c i p r i c e r c a , s v i l u p p o e i n n o v a z i o n e ” e s .m .i .;

VISTO i l Re g o l a m e n t o (UE) n . 651/2014 d e l l a Co m m i s s i o n e , d e l 17 g i c a t e g o r i e d i a i u t i c o m p a t i b i l i c o n i l m e r c a t o i n t e r n VISTA l a c o m u n i c a z i o n e d e l l a Co m m i s s i o n e 2016/C 262/01 s u l l a a l l 'a r t i c o l o 107, p a r a g r a f o 1, d e l t r a t t a t o s u l f u n z i o n a m VISTA l a Co m u n i c a z i o n e d e l l a Co m m i s s i o n e d e l 19 m a r z o 2020, C l e m i s u r e d i a i u t o d i St a t o a s o s t e g n o d e l l 'e c o n o m i a n e r e t t i f i c a t a a t t r a v e r s o l a c o m u n i c a z i o n e d e l 00 x x x x x x t e m p o r a n e o p e r l e m i s u r e d i a i u t o d i St a t o a s o s t e g n o d 19 e m o d i f i c a d e l l 'a l l e g a t o d e l l a c o m u n i c a z i o n e d e l l a d e g l i a r t i c o l i 107 e 108 d e l t r a t t a t o s u l f u n z i o n a m e n t o

a l l 'e s p o r t a z i o n e a b r e v e t e r m i n e ” ;

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2.1.1 - p r o o f o f c o n c e p t , 2.1.2 – t u m o r i e m a l a t t i e r a r e e 2.1.3 r i p a r t i z i o n e d e g l i i n t e r v e n t i d i i n v e s t i m e n t o d e l l a Na z i o n a l e d i Ri p r e s a e Re s i l i e n z a r e l a t i v o a l l 'i n n o v a z s a n i t a r i o n a z i o n a l e e a l p o t e n z i a m e n t o d e l s i s t e m a d VISTO i l I° a v v i s o p u b b l i c o p e r l a p r e s e n t a z i o n e e s e l

n e l l ’ a m b i t o d e l PNRR, p u b b l i c a t o s u l s i t o w e b d e l Mi n i s t u f f i c i a l e d e l l a Re p u b b l i c a i t a l i a n a , s u l l e s e g u e n t i t c o n e s c l u s i o n e d e i t u m o r i r a r i , Ma l a t t i e Cr o n i c h e n o n s a n i t a r i e s o c i o -a s s i s t e n z i a l i (Fa t t o r i d i r i s c h i o e p VISTO i l d e c r e t o d i r e t t o r i a l e n . 27 d e l 2 n o v e m b r e 0000, x x x x x x x x i l a n c i o i n d a t a 18 n o v e m b r e 2022, c o n i l q u a l e è s t a t a a PNRR- Mi s s i o n e 6 - Co m p o n e n t e 2 - In v e s t i m e n t o 2.1, a f f e r e n t i Co n c e p t , Ma l a t t i e r a r e , Ma l a t t i e c r o n i c h e n o n t r a s m i s

a s s i s t e n z i a l i (t e m a t i c h e : Fa t t o r i d i r i s c h i o e p r e v e n i l q u a l e s i è p r o c e d u t o a d i n d i v i d u a r e i l So g g e t t o a t t VISTO l ’ a r t . 7 d e l d e c r e t o m i n i s t e r i a l e 8 a p r i l e 2015, r e c a n t n o n g e n e r a l e d e l Mi n i s t e r o d e l l a s a l u t e , o v e v e n g o n o i

g e n e r a l e d e l l a r i c e r c a e d e l l ’ i n n o v a z i o n e i n s a n i t à u f f i c i 3 e 4 d e l l a s t e s s a ;

VISTO i l d e c r e t o d i r e t t o r i a l e d e l 1° m a r z o 2022, r e g i s t r a t m a r z o 2022, a l n . 247, c o n i l q u a l e i l Do t t . Ga e t a n o Gu x x x x x x x è d e l p o t e r e d i s p e s a e l ’ o r d i n e d i s e r v i z i o c o n i l q u a l e p e r i p r o g e t t i r i s u l t a t i v i n c i t o r i n e l b a n d o PNRR;

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1. Le p r e m e s s e s o n o p a r t e i n t e g r a n t e e s o s t a n z i a l e d e l l

2. Fa a l t r e s ì p a r t e i n t e g r a n t e e s o s t a n z i a l e d e l l a p r e p r o g e t t o d i r i c e r c a , i c u i c o n t e n u t i s o n o d e f i n i t i e c o n d i v i s i o n e d e l l e p a r t i , s e n z a n e c e s s i t à d i e s p r e s

Ar t . 2 So g g e t t o a t t u a t o r e /b e n e f i c i a r i o e Pr i n c i

Il So g g e t t o a t t u a t o r e -b e n e f i c i a r i o e i l Pr i n c i p a l In p r o g e t t o i n q u e s t i o n e e d e l l a r e g o l a r i t à d e l l e r e l a t i

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d e l i v e r e d b y Li p i d Na n o , pn ae rl TlIC’l ae ms b (iTAt CoTICd )e l l a r e a l i z z a p r e v i s t i d a l PNRR, Mi s s i o n e 6 – Co m p o n e n t e 2 – In v e s t i m e n

2. La p r e s e n t e Co n v e n z i o n e d e f i n i s c e , t r a l ’ a l t r o , g l i o e q u e l l e d i p a g a m e n t o .

3. Il s o g g e t t o a t t u a t o r e -b e n e f i c i a r i o e i l Pr i n c i p a l In q u a n t o r i p o r t a t o n e l p r o g e t t o p r e s e n t a t o p a r t e i n t Mi n i s t e r o e i n o t t e m p e r a n z a a q u a n t o p r e v i s t o d a l l ’

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1. La p r e s e n t e c o n v e n z i o n e h a l a d u r a t a d i 24 m e s i p r o r o g p r e v i s t o d a l s u c c e s s i v o a r t i c o l o 11.

2. L’ a t t i v i t à d i r i c e r c a , d a s v o l g e r s i n e l l ’ a r c o t e m p o i n i z i o i m p r o r o g a b i l m e n t e e n t r o e n o n o l t r e i l 20 m a g g c o n n o t a s o t t o s c r i t t a d i g i t a l m e n t e d a l p r o p r i o r a p r i c e r c a c h e d e v e e s s e r e t r a s m e s s a a l m e n o 30 g i o r n i d o c u m e n t a z i o n e d i c u i a l s u c c e s s i v o c o m m a 4.

3. Il So g g e t t o b e n e f i c i a r i o e n t r o e n o n o l t r e 15 g i o r n i d a Mi n i s t e r o p e r l a s o t t o s c r i z i o n e p r o v v e d e a l l a r e s r a p p r e s e n t a t e e c o n t r o f i r m a t a d a l Pr i n c i p a l In v e s t i a c c o m p a g n a t a d a l l a c o m u n i c a z i o n e d e l c o d i c e CUP MAST s i n g o l e Un i t à o p e r a t i v e . Le p a r t i r i c o n o s c o n o c h e i l

d a l f i n a n z i a m e n t o i n c a s o d i i n a d e m p i e n z a d e l l a p r e

4. Il So g g e t t o b e n e f i c i a r i o , e n t r o e n o n o l t r e 30 g i o r n i p r 2 d e l p r e s e n t e a r t i c o l o , p e n a l a d e c a d e n z a d a l f i n a s o t t o s c r i t t a d i g i t a l m e n t e i n m a n i e r a c o n g i u n t a d In v e s t i g a t o r d e l l a r i c e r c a - l a s e g u e n t e d o c u m e n t a z f i n e d i a u t o r i z z a r e l ’ a v v i o d e l p r o g e t t o :

a ) l a d i c h i a r a z i o n e d a p a r t e d e l l e g a l e r a p p r e s e d i c h i a r i c h e i l p r o g e t t o i n q u e s t i o n e o p a r t i s

f i n a n z i a m e n t i p u b b l i c i a f a v o r e d e l l ’ En t e a t t u e c h e , i n o g n i c a s o , s a r à p o s t a i n e s s e r e o g n i i f i n a n z i a m e n t o ;

b ) l a d i c h i a r a z i o n e d a p a r t e d e l l e g a l e r a p p r e s e n u n i t à o p e r a t i v a p a r t e c i p a n t e c o n c u i s i d i c h i p r o g e t t o i n q u e s t i o n e o p e r p a r t i s i g n i f i c a t f i n a n z i a m e n t i p u b b l i c i a f a v o r e d e l l ’ Un i t à o p e o p e r a t i v e e l e n c a t i n e l l a p r o p o s t a p r o g e t t u a l

i n i z i a t i v a v o l t a a d e v i t a r e i l d o p p i o f i n a n z i a c ) l a d i c h i a r a z i o n e d a p a r t e d e g l i En t i c h e s v o l g o

r e s p o n s a b i l i d i a c c e t t a z i o n e d e i t e r m i n i d e l l d ) l a d i c h i a r a z i o n e c o n l a q u a l e i l So g g e t t o b e n e f

s v o l g e r à l a p r o p r i a a t t i v i t à d i r i c e r c a , p e r l p r o g e t t o , e s c l u s i v a m e n t e p r e s s o l a p r o p r i a s e d m e d e s i m o , c o n t r o f i r m a t a d a l l ’ i n t e r e s s a t o ;

e ) i l p a r e r e p o s i t i v o d e l Co m i t a t o e t i c o c o m p e t e n d e l d e c r e t o l e g i s l a t i v o n . 26 d e l 4 m a r z o 2014 r i g u a r p r e v i s t i ;

f ) l a c o m u n i c a z i o n e d e l c o d i c e CUP d e l l e s i n g o l e Un i l c o d i c e f i s c a l e d e i s o g g e t t i d e s i g n a t i a o p e f o r m a t e x c e l c h e v e r r à c o n d i v i s o d a p a r t e d e d e l l ’ i n n o v a z i o n e i n s a n i t à c h e d o v r à e s s e r e r e

g ) l a t r a d u z i o n e i n l i n g u a i t a l i a n a d e l l a p r o p o s t a l l a v e r s i o n e i n i n g l e s e a l l e g a t a a l l a p r e s e n t

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b a s e a l l a t a b e l l a a l l e g a t a a l d e c r e t o m i n i s t e r i a l e c o n c e r n e n t e l a r i p a r t i z i o n e d e g l i i n t e r v e n t i d i i n In v e s t i m e n t o 2.1 d e l Pi a n o Na z i o n a l e d i Ri p r e s a e Re s i l i a l l a d i g i t a l i z z a z i o n e d e l Se r v i z i o s a n i t a r i o n a z i o b i o m e d i c a .

6. La p r e s e n t a z i o n e d e l l a r i c h i e s t a d i p a g a m e n t o d e l l a l e m o d a l i t à p r e v i s t e d a l l ’ a r t . 13, p a r a g r a f o 13.1 d e l b a n d e l l a d o c u m e n t a z i o n e a s u p p o r t o n e l s i s t e m a Re Gi S, e n d a p a r t e d e l Mi n i s t e r o d e l l ’ a p p r o v a z i o n e d e l l a r e l

7. La p r e s e n t a z i o n e d e l l a r i c h i e s t a d i p a g a m e n t o f i n a s u c c e s s i v a m e n t e a l l ’ i n v i o e n t r o 30 g i o r n i d a l l a d a p r o r o g a t a s e c o n d o i t e r m i n i d e l l a p r e s e n t e c o n v e n z i r e n d i c o n t a z i o n e e c o n o m i c a c o m p l e s s i v a d e l p r o g e t t d a p a r t e d e l Mi n i s t e r o d e l l ’ a p p r o v a z i o n e d e l l a r e l

8. Il m a n c a t o a d e m p i m e n t o d i q u a n t o p r e v i s t o d a i c o m m i 2 a r e a l i z z a r e i l p r o g e t t o e c o m p o r t a l a d e c a d e n z a

f i n a n z i a m e n t o .

Ar t . 5 Ob b l i g h i d e l So g g e t t o a t t u a t o r e -b e n e f i c i a r i o

1. Co n l a s o t t o s c r i z i o n e d e l l a p r e s e n t e Co n v e n z i o n e , i In v e s t i g a t o r , p e r q u a n t o d i c o m p e t e n z a , s i o b b l i g a n o

1) a s s i c u r a r e i l r i s p e t t o d i t u t t e l e d i s p o s i z i o n i p r p a r t i c o l a r e r i f e r i m e n t o a q u a n t o p r e v i s t o d a l Re g . c o n v e r t i t o c o n m o d i f i c a z i o n i d a l l a L. 29 l u g l i o 2021, n .

2) g a r a n t i r e i l r i s p e t t o d i e v e n t u a l i p r e v i s i o n i n o r Mi n i s t e r o d e l l a s a l u t e , d a l Mi n i s t e r o d e l l ’ e c o n o m o v v e r o d a a l t r i s o g g e t t i c o i n v o l t i n e l l ’ a t t u a z i o n a n c h e s u c c e s s i v a m e n t e a l l a s o t t o s c r i z i o n e d e l l a p

3) a s s i c u r a r e l ’ a d o z i o n e d i m i s u r e a d e g u a t e v o l t e a r s e c o n d o q u a n t o d i s c i p l i n a t o n e l Re g o l a m e n t o f i n a n d e l Re g o l a m e n t o (UE) 2021/241, i n p a r t i c o l a r e i n m a t e r i a d

i n t e r e s s i , d e l l e f r o d i , d e l l a c o r r u z i o n e , d e l d o p p i f o n d i c h e s o n o s t a t i i n d e b i t a m e n t e a s s e g n a t i ;

4) r i s p e t t a r e , a p e n a d i s o s p e n s i o n e o r e v o c a d e l f i n p r i n c i p i o d i “ n o n a r r e c a r e d a n n o s i g n i f i c a t i v o ” d e l l ’ a r t i c o l o 17 d e l Re g o l a m e n t o (UE) 2020/852, i p r i n c i p i

l ’ a l t r o , i l p r i n c i p i o d e l c o n t r i b u t o a l l ’ o b i e t t i v p r o d u c e n d o d a t i r e l a t i v i a i d e s t i n a t a r i e f f e t t i

r e l a z i o n e a g l i a r t i c o l i 2, 3, p a r a g r a f o 3, d e l TUE, 8, 10, 19 e d e i d i r i t t i f o n d a m e n t a x x x x x x ’ Un i o n e e u r o p e a ), l ’ o e d e v e n t u a l i u l t e r i o r i r e q u i s i t i e c o n d i z i o n a l i t Co n v e n z i o n e ;

5) a d o t t a r e p r o p r i e p r o c e d u r e i n t e r n e , a s s i c u r a n d o l i n d i c a t o d a l Mi n i s t e r o n e l l a d e s c r i z i o n e d e l l e f u

6) d a r e p i e n a a t t u a z i o n e a l p r o g e t t o c o s ì c o m e i l l u f i n a n z i a m e n t o d a l Mi n i s t e r o , g a r a n t e n d o l ’ a v v i o

i n c o r r e r e i n r i t a r d i a t t u a t i v i e c o n c l u d e r e i l p r o r i s p e t t o d e l l a t e m p i s t i c a p r e v i s t a d a l r e l a t i v o Mi n i s t e r o l e e v e n t u a l i m o d i f i c h e a l p r o g e t t o ;

7) a s s i c u r a r e i l r i s p e t t o d e l l a n o r m a t i v a v i g e n t e s u

8) a s s i c u r a r e i l r i s p e t t o d e i c r i t e r i d i a m m i s s i b i l d a l l ’ Av v i s o p e r l e v a r i e v o c i d i c o s t o , c h e s a r a n n o c a l n e t t o d i e v e n t u a l i e c o n o m i e r i s c o n t r a t e s u l f i n d o p o v e r i f i c a d a p a r t e d e l Mi n i s t e r o ;

9) g a r a n t i r e , n e l c a s o i n c u i s i f a c c i a r i c o r s o a l l e p r d e c r e t o l e g i s l a t i v o n . 50/2016 e s .m .i .; r i s p e t t a r e , i n c a l l ’ Am m i n i s t r a z i o n e , l a c o n f o r m i t à a l l a p e r t i n e n t

e v e n t u a l i s p e c i f i c h e c i r c o l a r i /d i s c i p l i n a r i c h e p

10) i n d i v i d u a r e e v e n t u a l i f a t t o r i c h e p o s s a n o d e t e r m i s u l l a t e m p i s t i c a a t t u a t i v a e d i s p e s a d e f i n i t a n e l s t e s s i ;

11) m i t i g a r e e g e s t i r e i r i s c h i c o n n e s s i a l p r o g e t t o

a l l ’ a n d a m e n t o g e s t i o n a l e e d a l l e c a r a t t e r i s t i c h e

12) e f f e t t u a r e i c o n t r o l l i o r d i n a r i d i g e s t i o n e e d i

n o r m a t i v a v i g e n t e , e l e v e r i f i c h e s u l c o n f l i t t o d i i d a l l a n o r m a t i v a a n t i r i c i c l a g g i o (“ t i t o l a r e e f f e t t

13) u t i l i z z a r e i l s i s t e m a i n f o r m a t i c o “ Re Gi S, f i n a l i z z a e l e t t r o n i c o i d a t i p e r c i a s c u n a o p e r a z i o n e n e c e s s

f i n a n z i a r i a , l a v e r i f i c a e l ’ a u d i t , s e c o n d o q u a n t o (UE) 2021/241 e t e n e n d o c o n t o d e l l e i n d i c a z i o n i c h e v e r r a i l t r a m i t e d e l Mi n i s t e r o ;

14) c a r i c a r e s u l p o r t a l e Wo r k f l o w d e l l a Ri c e r c a e n e l s c i e n t i f i c a s u l l o s t a t o d i a v a n z a m e n t o d e l p r o g e t s t e s s o ;

15) c a r i c a r e s u l s i s t e m a i n f o r m a t i v o “ Re Gi S” l a d o c u

s v o l g i m e n t o d e i c o n t r o l l i o r d i n a r i p r e v i s t i d a l l a e s p l e t a t e p e r l ’ a g g i u d i c a z i o n e d e g l i e v e n t u a l i a p r i c h i e s t a d a l l e Am m i n i s t r a z i o n i c e n t r a l i d e p u t a t

16) g a r a n t i r e l a c o r r e t t e z z a , l ’ a f f i d a b i l i t à e l a c o n l ’ a l i m e n t a z i o n e d e l s i s t e m a i n f o r m a t i v o “ Re Gi S” d e a i CUP d e l l e s i n g o l e Un i t à o p e r a t i v e s u l l ’ a v a n z a m e n c h e c o m p r o v a n o i l c o n s e g u i m e n t o d e g l i o b i e t t i v i

i n d i c a t o r i a d o t t a t i p e r l e m i l e s t o n e s e i t a r g e t d e a l m e n o b i m e s t r a l e d e l l e s p e s e (n e l t e r m i n e m a s s i m o b i m e s t r e ) n e l p o r t a l e Wo r k f l o w d e l l a Ri c e r c a e s u l

d o c u m e n t a z i o n e p r o b a t o r i a p e r t i n e n t e , s a l v o d i v e r

17) r i s p e t t a r e l ’ o b b l i g o d i i n d i c a z i o n e d e l CUP s u t u t p r o g e t t o e s u i d o c u m e n t i c o l l e g a t i a l l e r e l a t i v e p

18) f o r n i r e t u t t e l e i n f o r m a z i o n i r i c h i e s t e r e l a t i v a

s p e s e r e n d i c o n t a t e c o n f o r m e m e n t e a l l e p r o c e d u r e e

19) g a r a n t i r e l a c o n s e r v a z i o n e d e l l a d o c u m e n t a z i o n e p a s s i c u r a r e l a c o m p l e t a t r a c c i a b i l i t à d e l l e o p e r a z 4, d e l D.L. n . 77 d e l 31 m a g g i o 2021, c o n v e r t i t o c o n m o d i f i c a z d i v e r s e f a s i d i c o n t r o l l o e v e r i f i c a p r e v i s t e d a l s e s s e r e m e s s i p r o n t a m e n t e a d i s p o s i z i o n e s u r i c h i

i n t e r v e n t o PNRR, d e l Se r v i z i o c e n t r a l e p e r i l PNRR d Co m m i s s i o n e e u r o p e a , d e l l ’ OLAF, d e l l a Co r t e d e i Co n t i (EPPO) e d e l l e c o m p e t e n t i Au t o r i t à g i u d i z i a r i e n a z i o n Co r t e d e i c o n t i e l 'EPPO a e s e r c i t a r e i d i r i t t i d i c u i

f i n a n z i a r i o (UE; XXXXXXX) 1046/2018;

20) f a c i l i t a r e l e v e r i f i c h e d e l l ’ Uf f i c i o c o m p e t e n t e p Co m m i s s i o n e e u r o p e a e d i a l t r i o r g a n i s m i a u t o r i z z a a t t r a v e r s o c o n t r o l l i i n l o c o ;

21) a s s i c u r a r e c h e l e s p e s e d e l Pr o g e t t o d i r i c e r c a n o f i n a n z i a m e n t i , c o n t r i b u t i o a g e v o l a z i o n i a v a l e r e d e l d o p p i o f i n a n z i a m e n t o );

22) g a r a n t i r e l a d i s p o n i b i l i t à d e i d o c u m e n t i g i u s t i f i c o s ì c o m e p r e v i s t o a i s e n s i d e l l ’ a r t i c o l o 9 p u n t o 4 d i n l e g g e 29 l u g l i o 2021, n . 108;

23) p r e d i s p o r r e i p a g a m e n t i s e c o n d o l e p r o c e d u r e s t a f i n a n z i a r i o e c r o n o g r a m m a d i s p e s a a p p r o v a t o , i n s e e v e n t u a l m e n t e d i 6 m e s i ) n e l p o r t a l e Wo r k f l o w d e l l a r e l a t i v i d o c u m e n t i r i f e r i t i a l l e p r o c e d u r e e i g i u o r d i n a r i d i l e g a l i t à e a i c o n t r o l l i a m m i n i s t r a t i a p p l i c a b i l e , n e l r i s p e t t o d i q u a n t o p r e v i s t o d a l l ’ d e l d e c r e t o l e g g e n . 77 d e l 31/05/2021, c o n v e r t i t o i n l e g g e 2

24) a s s i c u r a r e c h e t u t t e l e s p e s e r e n d i c o n t a t e s i a n o s p r o g e t t o e c h e g l i e v e n t u a l i p a g a m e n t i p e r f a t t u r e s i a n o c o m p l e t a t e e n t r o i 30 g i o r n i s u c c e s s i v i a l l a c a r i c a m e n t o s u l s i s t e m a d i r e n d i c o n t a z i o n e Re Gi S;

25) i n o l t r a r e , a l l o s c a d e r e d e i 12 e 24 m e s i (p r o r o g a b i l i p a g a m e n t o a l Mi n i s t e r o t r a m i t e i l p o r t a l e Wo r k f l “ Re Gi S” c o n a l l e g a t a l a r e n d i c o n t a z i o n e d e t t a g l i a c o n t r i b u t o a l p e r s e g u i m e n t o d e l l e m i l e s t o n e s e d e i u n i t a m e n t e a i d o c u m e n t i g i u s t i f i c a t i v i a p p r o p r i a d i s p o s i t i v i a t t u a t i v i ;

26) g a r a n t i r e l ’ u t i l i z z o d i u n c o n t o c o r r e n t e d e d i c a l ’ a d o z i o n e d i u n a c o n t a b i l i t à s e p a r a t a o d i u n ’ a p p t u t t e l e t r a n s a z i o n i r e l a t i v e a l p r o g e t t o a l f i n e d d e l PNRR;

27) a s s i c u r a r e , d i r e t t a m e n t e o a t t r a v e r s o l e Is t i t u z i o d i r i c e r c a , l ’ a n t i c i p a z i o n e d e l l e s o m m e n e c e s s a r i

28) p a r t e c i p a r e , o v e r i c h i e s t o , a l l e r i u n i o n i c o n v o c a t

29) g a r a n t i r e , a n c h e a t t r a v e r s o l a t r a s m i s s i o n e d i r e l p r o g e t t o , c h e i l Mi n i s t e r o r i c e v a t u t t e l e i n f o r m a z l ’ e l a b o r a z i o n e d e l l e r e l a z i o n i a n n u a l i d i c u i a l l n o n c h é q u a l s i a s i a l t r a i n f o r m a z i o n e e v e n t u a l m e n t

30) c o n s e g u i r e i l r a g g i u n g i m e n t o d e g l i o b i e t t i v i d e l l a d o t t a t i p e r l e m i l e s t o n e s e i t a r g e t d e l l a m i s u r a Mi n i s t e r o , l e i n f o r m a z i o n i n e c e s s a r i e p e r l a p r e d i d i t a r g e t e m i l e s t o n e s e d e l l e r e l a z i o n i e d o c u m e n t

31) g a r a n t i r e i l r i s p e t t o d e g l i o b b l i g h i i n m a t e r i a d i d e l Re g o l a m e n t o (UE) 2021/241 i n d i c a n d o n e l l a d o c u m e n t a f i n a n z i a t o n e l l ’ a m b i t o d e l PNRR, c o n e s p l i c i t o r i f e

e u r o p e a e a l l ’ i n i z i a t i v a Ne x t Ge n e r a t i o n EU (a d e s . u e u r o p e a – Ne x t Ge n e r a t i o n EU – PNRR M6C2 - In v e s t i m e n t o 2.1 d e l l a r i c e r c a b i o m e d i c a d e l SSN” ), r i p o r t a n d o n e l l a d e u r o p e a e f o r n i r e u n ’ a d e g u a t a d i f f u s i o n e e p r o m o z i i n l i n e a c o n q u a n t o p r e v i s t o d a l l a St r a t e g i a d i Co m u

32) f o r n i r e i d o c u m e n t i e l e i n f o r m a z i o n i n e c e s s a r i e s t a b i l i t e d a i Re g o l a m e n t i c o m u n i t a r i e d a l Mi n i s t e

33) g a r a n t i r e u n a t e m p e s t i v a d i r e t t a i n f o r m a z i o n e a g l s u l l ’ a v v i o e l ’ a n d a m e n t o d i e v e n t u a l i p r o c e d i m e a m m i n i s t r a t i v o c h e d o v e s s e r o i n t e r e s s a r e l e o p e

i r r e g o l a r i t à , l e f r o d i , i c a s i d i c o r r u z i o n e e d i c f i n a n z i a m e n t o , r i s c o n t r a t i a s e g u i t o d e l l e v e r i f i

n	e	l	r i s p e t t o d e l l e p r o c e d u r e a d o t t a t e d a l l o s t e s s
d	e	l	Re g o l a m e n t o (UE) 2021/2041;
34) g	a	r	a n t i r e c h e i l Mi n i s t e r o r i c e v a a t t r a v e r s o i l s i
l	’	a	g g i o r n a m e n t o d e l l ’ i n d i c a t o r e c o m u n e n . 8 “ Ri c e
b	e	n	e f i c i a r i d i u n s o s t e g n o ” , r i c o n d u c i b i l e a l l a m i
a	i	s	e n s i d e l l ’ a r t . 3, c o m m a 3, d e l Re g o l a m e n t o d e l e g a t o
s	e	t	t e m b r e 2021 c h e i n t e g r a i l r e g o l a m e n t o (UE) 2021/241 d e l
c	h	e	i s t i t u i s c e i l d i s p o s i t i v o p e r l a r i p r e s a e l a
l	’	a	g g i o r n a m e n t o d e g l i i n d i c a t o r i c o m u n i h a l u o g o
p	e	r	i o d o d i r i f e r i m e n t o c o p r e l ’ i n t e r o p e r i o d o d i a
d	e	l	c a s o , f i n o a l l e r i s p e t t i v e d a t e l i m i t i d e l 31 d i c

Ar t . 6 Pr o c e d u r a d i m o n i t o r a g g i o e r e n d i c o n t a z i o n e

1. Il Mi n i s ct oe nr lo a p r e s e n t e c o n v e n z i o n e r a p p r e s e n t a a l l a s c i e n t i f i c o s a r à s v o l t o d a l l a Di r e z i o n e d e l l a Ri c e r i s p e t t o a l l a r e n d i c o n t a z i o n e d e l l e s p e s e s a r a n n o

i n t e r v e n t pi r de es ls oPNiRRl Mi n i s t e r o d e l l a s a l u t e .

2. Il So g g e t t o a t t u a t o r e -b e n e f i c i a r i o , s e c o n d o l e i n d i c a l m e n o b i m e s t r a l e , e n t r o 10 g i o r n i s u c c e s s i v i a l l ’ s u l l ’ a v a n z a m e n t o f i n a n z i a r i o , f i s i c o e p r o c e d u r a l i m p l e m e n t a r e t a l e s i s t e m a c o n l a d o c u m e n t a z i o n e a f f i d a m e n t o e a c i a s c u n a t t o g i u s t i f i c a t i v o d i s p e s d e i c o n t r o l l i a m m i n i s t r a t i v o -c o n t a b i l i a n o r m a d e l

d i m i s s i o n e p e r l 'a t t u a z i

po rn ee sdMsei gonl ii i sl it

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eo rdveel nl t a i s da el l u Pt NeR

3. Il So g g e t t o a t t u a t o r e -b e n e f i c i a r i o , a l l o s c a d e r e d e i d e v e t r a s m e t t e r e i d a t i s u l l ’ a v a n z a m e n t o t e c n i c o -s d e l l a Ri c e r c a e i l s i s t e m a “ Re Gi S” c o r r e d a t a d i d o c u m e d i a f f i d a m e n t o e a c i a s c u n a t t o g i u s t i f i c a t i v o d i l ’ e s p l e t a m e n t o d e i c o n t r o l l i a m m i n i s t r a t i v o -c o n t a p r o g e t t o .

4. Il So g g e t t o a t t u a t o r e -b e n e f i c i a r i o , p e r t a n t o , d o v r à i e v e n t u a l m e n t e d i 6 m e s i ) t r a m i t e i l p o r t a l e Wo r k f l o w l a r i c h i e s t a r e n d i c o n t a z i o n e d e l l e s p e s e v o l t e a s e s s e r e f o r m a l m e n t e t r a s m e s s e a l l ’ Un i t à d i Mi s s i o n e s p e s e e f f e t t i v a m e n t e s o s t e n u t e n e l p e r i o d o d i r i f e i n t e r v e n t o /p r o g e t t o c o n s p e c i f i c o r i f e r i m e n t o a l l e e s s e r e c o r r e d a t a d a l l a d o c u m e n t a z i o n e s p e c i f i c a t Mi n i s t e r o .

5. Le s p e s e i n c l u s e n e l l e r i c h i e s t e d i p a g a m e n t o d e l S o p e r a z i o n i e s t r a t t e a c a m p i o n e , s o n o s o t t o p o s t e , p e r v e r i f i c h e , s e d e l c a s o a n c h e i n l o c o d a p a r t e d e l l e s t

6. Ne l l o s p e c i f i c o , l ’ Un i t à d i m i s s i o n e pd ee Mrl i ln 'ai

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zo i doen

Sa l u t e e e v e n t u a l i a l t r e a m m i n i s t r a z i o n i c o i n v o l t e s u l l e p r o c e d u r e , s u l l e s p e s e e s u i t a r g e t i n c o n f o r m i (UE) 2021/241 a l f i n e d i g a r a n t i r e l a t u t e l a d e g l i i n t e r e

i n d i v i d u a z i o n e e r e t t i f i c a d i f r o d i , d i c a s i d i c o r r u s o m m e e r r o n e a m e n t e v e r s a t e o u t i l i z z a t e i n m o d o n o n

7. La Di r e z i o n e g e n e r a l e d e l l a Ri c e r c a e d i n n o v a z i o n e i m e r i t o l e f u n z i o n i d i v e r i f i c a t e c n i c o -s c i e n t i f i c a i n c o e r e n z a c o n l o s t a t o d i r e n d i c o n t a z i o n e d e l l e s p

Ar t . 7 Va l u t a z i o n e i n t e r m e d i a

1. Al l o s c a d e r e d e i 12 m e s i d a l l ’ i n i z i o d e l l ’ a t t i v i t à d d a t a l e t e r m i n e , i l So g g e t t o a t t u a t o r e -b e n e f i c i a r i o d e l l a r i c e r c a l a r e l a z i o n e i n t e r m e d i a s u l l o s t a t o d i g i t a l m e n t e d a l l e g a l e r a p p r e s e n t a n t e d e l So g g e t t

- c o n t e n e n t e l a d e s c r i z i o n e d e l l e a t t i v i t à p r o g e t t o p e r a t i v e , d a c u i r i s u l t i l o s t a t o a v a n z a m e n t o l a v o s e c o n d o q u a n t o r i p o r t a t o n e l p r o g e t t o a p p r o v a t o . Ta l Pr i n c i p a l In v e s t i g a t o r , c h e i l l u s t r i , n e l l a g l o b a l i d e s c r i z i o n e d e l l e a t t i v i t à r e a l i z z a t e d a e v e n t u a l

s u b c o n t r a e n t i .La r e l a z i o n e i n t e r m e d i a , p r e v i a v e r i f Ri c e r c a e d i n n o v a z i o n e i n s a n i t à , s a r à c a r i c a t a d a l In v e s t i g a t o r a l l ’ i n t e r n o d e l s i s t e m a i n f o r m a t i v o “

2. Il Mi n i s t e r o h a f a c o l t à , p r e v i a c o m u n i c a z i o n e p r e v e n l e p r o c e d u r e p e r l a s o s p e n s i o n e d e l f i n a n z i a m e n t o e d e g l i e v e n t u a l i i n t e r e s s i l e g a l i m a t u r a t i , q u a l o r

q u a n t o p r e v i s t o e n t r o i t e r m i n i d i c u i a l c o m m a 1 d e l p

3. La Di r e z i o n e g e n e r a l e d e l l a Ri c e r c a e d i n n o v a z i o n e c o m u n i c a z i o n e p r e v e n t i v a a l So g g e t t o a t t u a t o r e /b e n

m i s s i o n e p e r l 'a t t u a z i o n de ed le gmle id Mie i nsnti iemsrot x xx xx xx , x x xx xx x x XxXXx Rs

c o n d i z i o n i p e r n o n e r o g a r e l e s u c c e s s i v e q u o t e a r i g i u d i z i o i n o r d i n e a l l a r e l a z i o n e f i n a l e , q u a l o r a l

s i a c o n s i d e r a t a i d o n e a a d i m o s t r a r e c h e s i a n o s t a t i e m e r g a c h e e s s a s i a s t a t a c o n d o t t a n o n i n p i e n a c o n f o In t a l c a s o i l Mi n i s t e r o p o t r à p r o c e d e r e c o n i l r i m b t e r m i n i d i c u i a l l a p r e s e n t e c o n v e n z i o n e , c h e n o n c o

So g g e t t o a t t u a t o r e /b e n e f i c i a r i o e s o n e r a i l Mi n i s t e n e l l ’ e r o g a z i o n e d e l l e s o m m e s p e t t a n t i .

4. Il Mi n i s t e r o , p r e v i a c o m u n i c a z i o n e p r e v e n t i v a a l So g Co m i t a t o t e c n i c o s a n i t a r i o s e z . c ), u n d o s s i e r , q u a l o r a m i n i s t e r i a l e , n o n c o n s e n t a d i e s p r i m e r e u n c o m p i u t Co m i t a t o è v i n c o l a n t e p e r i l So g g e t t o b e n e f i c i a r i o a

Ar t . 8 Va l u t a z i o n e f i n a l e

1. Fa t t a s a l v a l ’ e v e n t u a l e c o n c e s s i o n e d i p r o r o g a d e l v e n t i q u a t t r o m e s i - e c o m u n q u e n o n o l t r e t r e n t a (30) g i r i c e r c a – a i f i n i d e l l ’ e r o g a z i o n e d e l s a l d o , i l So g d i g i t a l m e n t e d a l r a p p r e s e n t a n t e l e g a l e , t r a s m e t t

d o c u m e n t a z i o n e , r e d a t t a d a l Pr i n c i p a l In v e s t i g a t o r e

- l a r e l a z i o n e f i n a l e d e l l a r i c e r c a , c o n t e n e n t e q u c o f i n a n z i a t o r i , c h e d o c u m e n t i , p e r c i a s c u n a u n i t à i l p r o g e t t o a p p r o v a t o e g l i o b i e t t i v i r a g g i u n t i ;

- c o p i a d e i l a v o r i p u b b l i c a t i s u r i v i s t e i m p a t t a t e

- l a r e n d i c o n t a z i o n e d e l l e s p e s e s o s t e n u t e c o n i f o

- i n d i c a z i o n i d e l r e p o s i t o r y p u b b l i c o d o v e s o n o r e u t i l i z z a t i p e r l e p u b b l i c a z i o n i s c i e n t i f i c h e c o

- i l r i s p e t t o d e i c o s t i s o s t e n u t i r i s p e t t o a i v i n c o d a p a r t e d i i s t i t u z i o n i n e l l ’ a r e e d e l m e r i d i o n e

2. La r e n d i c o n t a z i o n e e c o n o m i c a d o v r à e s s e r e c o r r e d a t c h e c k l i s t d i v e r i f i c a d e i r e q u i s i t i m i n i m i d e l b a n i s c r i t t o a l l ’ Or d i n e d e i Do t t o r i Co m m e r c i a l i s t i e d E

i n p o s s e s s o d e i r e q u i s i t i r i c h i e s t i d a l l a Di r e t t i v a d e l 16 a p r i l e 2014 , c h e m o d i f i c a l a d i r e t t i v a 2006/43/CE r e l a t

e d e i c o n t i c o n s o l i d a t i , e d a l l a r e l a t i v a l e g i s l a l a r e g o l a r i t à a m m i n i s t r a t i v o -c o n t a b i l e d e l l e s p e s c o n f o r m i t à a l l a n o r m a t i v a d i r i f e r i m e n t o v i g e e d i t u t t i i r e q u i s i t i p r e v i s t i d a l l ’ Av v i s o e d a l l a n a z i o n a l i e d e u r o p e e i n m a t e r i a e l a c o n g r u e n z a c o n l

3. Tu t t a l a s o p r a r i c h i a m a t a d o c u m e n t a z i o n e d e v e e s s e r d e l l a r i c e r c a e i l s i s t e m a i n f o r m a t i c o “ Re Gi S” e s e c o d i m o n i t o r a g g i o e c o n o m i c o e u t i l i z z a n d o c o n g i u n t a d e l l a r i c e r c a , a d i s p o s i z i o n e d e i d e s t i n a t a r i i s t i t

t r a m i t e p o s t a e l e t t r o n i c a c e r t i f i c a t a (PEC) d a p a r t e d

4. La d o c u m e n t a z i o n e d i s u p p o r t o d e v e e s s e r e a d i s p o s i z v e r i f i c a d e l PNRR, p r e s s o i l So g g e t t o a t t u a t o r e /b e n e c u s t o d i a .

5. La Di r e z i o n e g e n e r a l e d e l l a Ri c e r c a e d i n n o v a z i o n e i a p p l i c a r e u n a d e c u r t a z i o n e p a r i a l 10% d e l l a r a t a d e l 1 d e l p r e s e n t e a r t i c o l o s i a t r a s m e s s a a l Mi n i s t e r o q u a r a n t e s i m o g i o r n o d a l l a d a t a d i c o n c l u s i o n e d e l p

6. Il Mi n i s t e r o p r o v v e d e a d a p p l i c a r e u n a d e c u r t a z i o n d o c u m e n t a z i o n e d i c u i a l c o m m a 1 d e l p r e s e n t e a r t i c c o m p r e s o t r a i l q u a r a n t u n e s i m o e i l c i n q u a n t e s i m o g

7. Il Mi n i s t e r o , p r e v i a c o m u n i c a z i o n e p r e v e n t i v a a l So g p e r l a s o s p e n s i o n e d e l f i n a n z i a m e n t o e l a c o n s e g u e r e c u p e r o d i t u t t e d e l l e s o m m e g i à e r o g a t e , a n c h e q u e l g r u p p o d e l l a r i c e r c a , c o m p r e n s i v e d e g l i i n t e r e s s i l c o m m a 1 d e l p r e s e n t e a r t i c o l o n o n s i a t r a s m e s s a a l Mi d i c o n c l u s i o n e d e l p r o g e t t o .

8. Il Mi n i s t e r o s i r i s e r v a l a f a c o l t à d i c h i e d e r e i n f o r So g g e t t o a t t u a t o r e /b e n e f i c i a r i o , c h e d e v e f o r n i r e r q u a l o r a :

- l a r e l a z i o n e f i n a l e n o n s i a c o n s i d e r a t a i d o n e a a i n c o n f o r m i t à a q u a n t o p r e v i s t o n e l p r o g e t t o e n e

- l a r e n d i c o n t a z i o n e r i s u l t i i n c o m p l e t a o i n c o n g r

9. Il Mi n i s t e r o p r o v v e d e r à a d e m e t t e r e l a v a l u t a z i o n e c a s o d i m a n c a t o o e s a u s t i v o r i s c o n t r o d a p a r t e d e l So a l p r e c e d e n t e c o m m a , i l Mi n i s t e r o c o m u n i c a a l So g g e t

o r d i n e a l l a r e l a z i o n e f i n a l e e c o n s e g u e n t e m e n t e i n c h i e d e r e l a r e s t i t u z i o n e d e l l e s o m m e g i à e r o g a t e , c o d i m a n c a t o r i s c o n t r o o p p u r e l a d d o v e d a l l ’ i s t r u t t o r s t a t i d i s a t t e s i g l i o b i e t t i v i d i c u i a l p r o g e t t o

10. Il Mi n i s t e r o , p r e v i a c o m u n i c a z i o n e p r e v e n t i v a a l So g Co m i t a t o t e c n i c o s a n i t a r i o s e z . c ) u n d o s s i e r , q u a l o r m i n i s t e r i a l e , n o n c o n s e n t a d i e s p r i m e r e u n c o m p i u t Co m i t a t o è v i n c o l a n t e p e r i l So g g e t t o b e n e f i c i a r i o a

Ar t . 9 Ve r i f i c a f i n a n z i a r i a p r e v e n t i v a Il So g g e t t o a t t u a t o r e -b e n e f i c i a r i o , a l f i n e d e l l ’ e r Mi n i s t e r o d e l l a s a l u t e – Un i t à d i m i s s i o n e p e r l ’ a t t s i s t e m a “ Re Gi S” l a r e n d i c o n t a z i o n e e c o n o m i c a c o r r e

a l c o m m a 2 d e l l ’ a r t i c o l o 8 d e l l a p r e s e n t e c o n v e n z i o n d i s o g g e t t i q u a l i f i c a t i a l l ’ Au d i t a l i v e l l o e u r o p e a l a c o m p l e t e z z a d e l l a d o c u m e n t a z i o n e i n b a s e a l l e d i q u e l l e e u r o p e e

Ar t . 10 Pr o c e d u r a d i p a g a m e n t o a l So g g e t t o b e n e

1. Le p r o c e d u r e d i e r o g a z i o n e d e i f o n d i s u r i c h i e s t a d a n t i c i p a z i o n e e a t i t o l o d i r i m b o r s o a l l ’ Un i t à d i

s p e c i f i c h e m o d a l i t à i n c o n f o r m i t à c o n q u a n t o i n d i c

- m a s s i m o 40% a l m o m e n t o d e l l a c o m u n i c a z i o n e , d a p a r

d e l l ’ a t t i v i t à d i r i c e r c a , a t i t o l o d i a n t i c i p a z i o

- q u o t a a r i m b o r s o p e r u n u l t e r i o r e p e r m a s s i m o u n 12° m e s e d a l l ’ i n i z i o d e l l e a t t i v i t à p r o g e t t u a l i , r e l a z i o n e s c i e n t i f i c a i n t e r m e d i a e d o p o l a s u a a p r i c h i e s t e d i p a g a m e n t o a t i t o l o d i r i m b o r s o p e r l b e n e f i c i a r i o , c o m e r i s u l t a n t i d a l s i s t e m a i n f o r

l e g g e 30 d i x x x x x x 0000, x , 000.

- q u o t a a r i m b o r s o r e s i d u a l e a s a l d o , a c o n c l u s i o n So g g e t t o a t t u a t o r e /b e n e f i c i a r i o d e l l a r e l a z i o n e c o n o m i c a , s u l l a b a s e d e l l a p r e s e n t a z i o n e d e l l a c o n c l u s i o n e d e l p r o g e t t o , i n c o e r e n z a c o n l e r i s a l l ’ a r t i c o l o 1, c o m m a 1043, d e l l a l e g g e 30 d i c e m b r e 2020

2. A g a r a n z i a d e l l a c o e r e n z a c o n l ’ i n i z i o d e l l ’ a t t i v i i m p e g n a a d a n t i c i p a r e l e r i s o r s e e c o n o m i c h e n e c e c o r r i s p o n d e r s i d a p a r t e d e l Mi n i s t e r o s i a n o i n r e g i

3. La d d o v e n o n v e n g a n o r i s p e t t a t i i t e r m i n i d i c u i a l l t e m p e s t i v a e r o g a z i o n e d e i f o n d i , i l So g g e t t o a t t u a t r e s p o n s a b i l i t à p e r e v e n t u a l i r i t a r d i n e l l ’ e r o g a z i

4. Al t e r m i n e d e l l e v e r i f i c h e l a Di r e z i o n e g e n e r a l e d e d e l l a Sa l u t e c o m u n i c h e r à d a l l ’ Un i t à d i m i s s di eo ln e p e

Mi n i s t e lr

eo rSai

ls uu lt

te a n z e d e l l e v e r i f i c h e p e r c o n s e n t i

p a g a m e n t i .

Ar t . 11 Va r i a z i o n i d e l p r o g e t t o e d e l p i a n o d e i

1. A p a r t i r e d a l 3° m e s e s u c c e s s i v o a l l ’ a v v i o d e l p r o g e p r o g e t t o , i l So g g e t t o a t t u a t o r e -b e n e f i c i a r i o , c o n n o Pr i n c i p a l In v e s t i g a t o r , t r a s m e s s a t r a m i t e i l p o r t a l “ Re Gi S” , p u ò p r o p o r r e v a r i a z i o n i a l p r o g e t t o , c o e r e n t

d i f o n d i t r a l e u n i t à o p e r a t i v e , p u r c h é n o n c o m p o r t i c a r i c o d e l Mi n i s t e r o , c h e d o v r a n n o e s s e r e a c c o l t e c o d i m o d i f i c a d e v e d i m o s t r a r e l e n e c e s s i t à s c i e n t i f i m o d i f i c a p r o p o s t a r i s p e t t o a l r a g g i u n g i m e n t o d e g l e f f i c a c i a s o l o d o p o l ’ a p p r o v a z i o n e d a p a r t e d e l Mi n p i a n o d e i c o s t i p e r i l CUP Ma s t e r e p e r i CUP d e l l e s i n g a t t u a t o r e -b e n e f i c i a r i o .

2. No n è c o n s e n t i t o a l d i f u o r i d e l p e r i o d o d i c u i a l c o m e v e n t u a l e n e c e s s i t à d i u n ’ u l t e r i o r e m o d i f i c a p r o g e s o l o d o p o 3 m e s i d a l l ’ a p p r o v a z i o n e d a p a r t e d e l Mi n i s t i p o l o g i a o v v e r o s i a s c i e n t i f i c a o e c o n o m i c a .

3. Il p i a n o d e i c o s t i , r i p o r t a t o n e l l a p r o p o s t a p r o g e t t t o t a l e d e l f i n a n z i a m e n t o a s s e g n a t o e a l r i p a r t o i n i i n d i c a t i v o p e r q u a n t o r i g u a r d a l a r i p a r t i z i o n e t r a d i t a l i c o s t i .

4. La d i s t r i b u z i o n e d e l l e s o m m e t r a l e d i v e r s e v o c i d i c è c o n s e n t i t a s o t t o l a r e s p o n s a b i l i t à d e l So g g e t t o a c h e d o v r à v e r i f i c a r e i l r i s p e t t o d e l l e p e r c e n t u a l i

5. Qu a l s i a s i p r o p o s t a e m e n d a t i v a d e v e e s s e r e a d e g u a t d o c u m e n t a r e c h e q u a n t o r i c h i e s t o r i s u l t i i n d i s p e n s a s u o t e m p o p r e f i s s a t i .

6. So l o d o p o l ’ a p p r o v a z i o n e d e l Mi n i s t e r o , i l s o g g e t a l l ’ a p p l i c a z i o n e d e l l e m o d i f i c h e d i c u i a l c o m m a

i n a d e m p i m e n t i a l p r e s e n t e a r t i c o l o i l Mi n i s t e r o h c o n v e n z i o n e , d a n d o n e c o m u n i c a z i o n e a l So g g e t t o a t t f i n a n z i a m e n t o , n o n c h é a l r e c u p e r o d i t u t t o l ’ i m p o r t

Ar t . 12 Pr o r o g a

1. Il t e r m i n e d e l l a r i c e r c a p u ò e s s e r e p r o r o g a t o d a l Mi n d i s c a d e n z a o r i g i n a l e , s o l o a s e g u i t o d i f o r m a l e , m o t d a l l e g a l e r a p p r e s e n t a n t e d e l So g g e t t o a t t u a t o r e -b t r a m i t e i l p o r t a l e Wo r k f l o w d e l l a r i c e r c a .

2. La r i c h i e s t a d i c u i a l c o m m a 1 p u ò e s s e r e a v a n z a t a s o l o t e r m i n e o v v e r o s i a d o p o 12 m e s i d a l l ’ a v v i o p r o g e t t o e c o n f o r m a l e e m o t i v a t a i s t a n z a d a p a r t e d e l So g g e t t o

c h e d i m o s t r i l e n e c e s s i t à s c i e n t i f i c h e a l l a b a s e r a g g i u n g i m e n t o d e g l i o b i e t t i v i p r o g e t t u a l i p r e v i s d e l Mi n i s t e r o .

Ar t . 13 Pr o p r i e t à e d i f f u s i o n e d e i r i s u l t a t

1. La p r o p r i e t à d e g l i s t u d i , d e i p r o d o t t i e d e l l e m e t o r e g o l a m e n t a t a d a l l a n o r m a t i v a v i g e n t e i n m a t e r i a , f i r m a t a r i e d e l p r e s e n t e a t t o , f e r m a r e s t a n d o l a p o s s Na z i o n a l e d i f r u i r n e , p r e v i a r i c h i e s t a a l l e p a r t i f i

2. Ne l c a s o i n c u i i l So g g e t t o a t t u a t o r e /b e n e f i c i a r i o i a n c h e p a r z i a l e , r e l a t i v o a l l a r i c e r c a i n q u e s t i o n e , a d e v e d a r n e p r e v e n t i v a c o m u n i c a z i o n e a l Mi n i s t e r o .

3. Il So g g e t t o a t t u a t o r e /b e n e f i c i a r i o s i i m p e g n a a g a r p r o g e t t o , a n c h e o n l i n e , s i a s u l w e b c h e s u i s o c i a l m e d

4. Qu a l s i a s i d o c u m e n t o p r o d o t t o , i v i c o m p r e s e l e p u b b l i o g g e t t o d e l l a p r e s e n t e c o n v e n z i o n e – p e r i q u a l i d e Mi n i s t e r o - d e v e c o n t e n e r e l ’ i n d i c a z i o n e c h e i l p r o u n ’ e s p l i c i t a d i c h i a r a z i o n e c h e r e c i t i "f i n a n z i a t o M6C2 - In v e s t i m e n t o 2.1 Va l o r i z z a z i o n e e p o t e n z i a m e n t

l ’ e m b l e m a d e l l ’ Un i o n e Eu r o p e a e d i l c o d i c e d e l p r o g e

5. I p r o d o t t i d i c u i a l p r e c e d e n t e c o m m a 4 d e v o n o e s s e r e r l ’ i m m e d i a t a f r u i z i o n e d a p a r t e d e l p u b b l i c o (a d e s e

d i p u b b l i c a z i o n e s c i e n t i f i c a p e r l a q u a l e s i a n e c e s p a g a m e n t o p e r l a c o n s u l t a z i o n e r e l a t i v a L’ e v e n t u a l s o l a p u b b l i c a z i o n e , s a r à o g g e t t o d i u n a p e n a l e p a r i a

6. Il Mi n i s t e r o n o n r i c o n o s c e l ’ e l e g g i b i l i t à d e i c o s t i p u b b l i c a z i o n i n o n s i f a c c i a e s p r e s s a m e n z i o n e d e l f c o d i c e p r o g e t t o .

7. Le p a r t i c o n v e n g o n o c h e i l Mi n i s t e r o p o s s a d a r e d i r e s i t o w e b , d e l l ’ e s t r a t t o d e l l a p r o p o s t a p r o g e t t u a l e s i n t e t i c a e d e l l e p u b b l i c a z i o n i s c i e n t i f i c h e d a e s

Ar t . 14 Ca s i d i r i d u z i o n e , s o s p e n s i o n e o r e v o c a d

a	.	m o d i	f	i	c h e i n g i u s t i f i c a t e a l l a c o m p o s i z i o n e d e l
b	.	m a n c	a	t	o r i s p e t t o d e i v i n c o l i p r e v i s t i d a l l ’ Av v i
c	.	m a n c	a	t	o r i s p e t t o d e g l i o b b l i g h i d i c u i a l l ’ a r t . 5
d	.	m a n c	a	t	o r a g g i u n g i m e n t o , n e i t e m p i a s s e g n a t i , d e
		s v o l	g	i	m e n t o d e l p r o g e t t o ;
e	.	m a n c	a	t	a o r i t a r d a t a p r e s e n t a z i o n e d e l l a r e l a z i
		r i c e	r	c	a ;
f	.	m a n c	a	t	a o r i t a r d a t a p r e s e n t a z i o n e - o l t r e i l c i n q
		p r o g	e	t	t o - d e l l a r e l a z i o n e f i n a l e d e l l a r i c e r c a e
		i f o n	d	i	m i n i s t e r i a l i ;
g	.	m o d i	f	i	c h e d e l p r o g e t t o o v a r i a z i o n i n e l l a d i s t r
		a u t o	r	i	z z a t e ;

1. Il Mi n i s t e r o p r o c e d e a d i c h i a r a r e l a s o s p e n s i o n e o r e c o n c o n s e g u e n t e e v e n t u a l e r e s t i t u z i o n e d e l l e s o m m m a t u r a t i , n e i s e g u e n t i c a s i :

2. Il Mi n i s t e r o a p p l i c a r i d u z i o n i f i n a n z i a r i e i n m i r i c o n o s c i m e n t o d e l l e s p e s e n e i s e g u e n t i c a s i :

a . m a n c a t o r i s p e t t o d e i c r i t e r i d i a m m i s s i b i l i t à d m a s s i m a l i p r e v i s t i p e r a l c u n e c a t e g o r i e d i s p p u b b l i c a z i o n i i n c u i n o n s i f a c c i a e s p r e s s a m e n z d e l PNRR e d e l c o d i c e p r o g e t t o ;

b . r i d u z i o n e f i n a n z i a r i a n e l l a m i s u r a d e l 5% d e l l a a t t u a t o r e /b e n e f i c i a r i o a l t e r m i n e d e l l e a t t i v i

c . r	i	d u	z	i	o	n e f i n a n z i a r i a n e l l a m i s u r a d e l 10% d e l l a
r	i	c e	r	c	a	e l a r e n d i c o n t a z i o n e d e l l e s p e s e s o s t e n
c	o	m p	r	e	s	o t r a i l t r e n t u n e s i m o e i l q u a r a n t e s i m o g
d . r	i	d u	z	i	o	n e f i n a n z i a r i a n e l l a m i s u r a d e l 20% d e l l a
r	i	c e	r	c	a	e l a r e n d i c o n t a z i o n e d e l l e s p e s e s o s t e n
c	o	m p	r	e	s	o t r a i l q u a r a n t u n e s i m o e i l c i n q u a n t e s
e f	. .	p r a r f r a p r p	r i t i i i t u e u	o d t v n d t b p b	g u u i a u u b o b	e t t o ; z i o n e f i n a n z i a r i a n e l l a m i s u r a d e l 5% d e l l ’ i a t o r e /b e n e f i c i a r i o a l t e r m i n e d e l l e a t t i v i s t e i m p a t t a t e a s e g u i t o d e l l o s v o l g i m e n t n z i a m e n t o o t t e n u t o n e l l ’ a m b i t o d e l PNRR e d e l z i o n e f i n a n z i a r i a n e l l a m i s u r a d e l 10% d e l l ’ i a t o r e /b e n e f i c i a r i o a l t e r m i n e d e l l e a t t i v l i c a t i s u r i v i s t e i m p a t t a t e a s e g u i t o d e l l o sp iu tb ob rl yi c o d o v e s o n o r e s i d i s p o n i b i l i i d a t i g l i c a z i o n i s c i e n t i f i c h e c o r r e l a t e .

r i v i s t e i m p a t t a t e a s e g u i t o d e l l o s v o l g i m e n t o d p u b b l i c a z i o n i p r o d o t t e r e c a n o l a m e n z i o n e d e l f e d e l c o d i c e p r o g e t t o ;

Ar t . 15 Ri s o l u z i o n e d i c o n t r o x x x x x x

0. Xx r q u a l s i a s i c o n t r o v e r s i a , i l So g g e t t o a t t u a t o r e -b e g e n e r a l e d e l l a r i c e r c a e d e l l ’ i n n o v a z i o n e i n s a n i t e v e n t u a l i p r o b l e m a t i c h e a l p a r e r e d i c o m p e t e n z a d e l i l Mi n i s t e r o . Le p a r t i , c o n l a s o t t o s c r i z i o n e d e l l a p r c h e s a r à e s p r e s s o d a l Co m i t a t o t e c n i c o s a n i t a r i o (C s c i e n t i f i c a d e l p r o g e t t o e l e e v e n t u a l i r i c a d u t e e c

2. Co n l a f i r m a d e l l a p r e s e n t e c o n v e n z i o n e i l Pr i n c i p a l c o m m a 1.

3. Qu a l o r a a s e g u i t o d e l l a v a l u t a z i o n e d e l CTS, d i c u i a c o n t r o v e r s i e , d i v e r s e d a q u e l l e d e l c o m m a 1, c h e d o v e s c o m p e t e n t e è i l Fo r o d i Ro m a .

Ar t . 16 Ri s o l u z i o n e p e r i n a d e m p i m e n t o

1. Il Mi n i s t e r o p o t r à a v v a l e r s i d e l l a f a c o l t à d i r i s o l a t t u a t o r e /b e n e f i c i a r i o n o n r i s p e t t i g l i o b b l i g h i

l ’ a s s o l v i m e n t o d a p a r t e d e l l o s t e s s o Mi n i s t e r o d e g l

Ar t . 17 Di r i t t o d i r e c e s s o

1. Il Mi n i s t e r o p o t r à r e c e d e r e i n q u a l u n q u e m o m e n t o d a g n e i c o n f r o n t i d e l So g g e t t o a t t u a t o r e /b e n e f i c i a r i o q d e l l e a t t i v i t à , i n t e r v e n g a n o f a t t i o p r o v v e d i m e n t i

s t i p u l a d e l l a p r e s e n t e Co n v e n z i o n e o n e r e n d a n o i m p

Ar t . 18 Co m u n i c a z i o n i e s c a m b i o d i i n f o r m a z i o

1. Ai f i n i d e l l a d i g i t a l i z z a z i o n e d e l l ’ i n t e r o c i c l o Mi n i s t e r o d e l l a s a l u t e d e v o n o a v v e n i r e a t t r a v e r s o i Wo r k f l o w d e l l a r i c e r c a , a d i s p o s i z i o n e d e l So g g e t t a t t r a v e r s o i l s i s t e m a m e s s o a d i s p o s i z i o n e d a l Mi “ Re Gi S” .

2. Il So g g e t t o a t t u a t o r e /b e n e f i c i a r i o a t t r a v e r s o i l p In v e s t i g a t o r d e v o n o f i r m a r e d i g i t a l m e n t e t u t t i g l i

Ar t . 19 Tr a c c i a b i l i t à d e i f l u s s i f i n a n z i a r

1. Le p a r t i s i i m p e g n a n o a l l ’ o s s e r v a n z a , p e r q u a n t o d i r a l l a t r a c c i a b i l i t à d e i f l u s s i f i n a n z i a r i d i c u i a l l

Ar t . 20 Pr o t e z i o n e d e i d a t i p e r s o n a l i

1. Ne l c o r s o d e l l ’ e s e c u z i o n e d e l l e a t t i v i t à o g g e t t o d e t r o v a r s i n e l l a c o n d i z i o n e d i d o v e r t r a t t a r e d a t i

d e l l ’ a l t r a Pa r t e , m o t i v o p e r c u i l e s t e s s e s i i m p e g n a p e r s o n a l i i n c o n f o r m i t à a l l e d i s p o s i z i o n i d i c u i a l

e d e l Co n s i g l i o , d e l 27 a p r i l e 2016, r e l a t i v o a l l a p r o t e t r a t t a m e n t o d e i d a t i p e r s o n a l i , n o n c h é a l l a l i b e r a 95/46/CE (Re g o l a m e n t o g e n e r a l e s u l l a p r o t e z i o n e d e i d a t a d e g u a m e n t o .

2. Le Pa r t i s i i m p e g n a n o a c o n d u r r e l e s u d d e t t e a t t i v i c o r r e t t e z z a , l i c e i t à , t r a s p a r e n z a e t u t e l a d e l l a r i e s c l u s i v o f i n e d i p e r s e g u i r e l e f i n a l i t à d i c u i a l l a d i l e g g e a l l o s t e s s o c o n n e s s i . Ta l i d a t i s a r a n n o t r a t

- a d o p e r a d i p r o p r i d i p e n d e n t i e /o c o l l a b o r a t o r i c h h a n n o l a n e c e s s i t à d i t r a t t a r l i , p e r l e s o l e f i n a l i n e c e s s a r i o a l l o r o c o n s e g u i m e n t o .

Ar t . 21 Ef f i c a c i a

1. La p r e s e n t e c o n v e n z i o n e , v i n c o l a n t e a l l 'a t t o d e l l a s e i l Pr i n c i p a l In v e s t i g a t o r , d i v e n t e r à e f f i c a c e p e r d e g l i o r g a n i d i c o n t r o l l o .

Ar t . 22 Di s p o s i z i o n i Fi n a l i

1. Pe r q u a n t o n o n p r e v i s t o d a l l a p r e s e n t e Co n v e n z i o n e r i f e r i m e n t o .

Le t t o , c o n f e r m a t o e s o t t o s c r i t t o c o n f i r m a d i g i t a l e , a s .m .i ..

Ro m a , (d a t a d e l l a s o t t o s c r i z i o n e c o m e q u e l l a d e l l ’ u l t i

p e r i l Mi n i s t e r o d e l l a s a l u t e

Do t t . Ga e t a n o Gu x x x x x x x Di r e t t o r e d e l l ’ Uf f i c i o 3

Di r e z i o n e g e n e r a l e d e l l a r i c e r c a e d e l l ’ i n n o v a z i o n e

p e r i l So g g e t t o a t Mt aura i t noar Cee,/bcr eoi

r a p p r e s e n t a n t e )

ndmeie fcl

iee CcfRiMi aMs rRcNi a5o9lC5e5H501I (Le g a l e

Pe r p r e s a v i s i o n e e d a c c e t t a z i o n e :

Il Pr i n c i p a l

GInEvNNeAsROt

Ci IgLIaBEt RoTOr , c-

o d i c e CfLBi GsNcR5a4Pl22eF839E

1 - General information

Project code: PNRR-POC-2022-12375713

PI / Coordinator: XXXXXXXXX XXXXXXX

Project topic: A) Proof of concept

Applicant Institution: Istituti fisioterapici ospitalieri - Istituto Regina Elena

Call section: Proposal title:

Proof of concept

TArgeting drug resistant melanoma with miCroRNAs delivered by Lipid NanoparTICles (TACTIC)

Duration in months: 24

MDC primary: Oncologia

MDC secondary:

Project Classification IRG:

Oncology 2 - Translational Clinical

Project Classification SS:

Drug Discovery and Molecular Pharmacology - DMP

Project Keyword 1:

Novel drug discovery: identification of molecular targets of antineoplastic agents that modulate signal translation, cell cycle, differentiation, apoptosis, and hormone signaling; mechanism of action of novel agents that lead to translation of these agents in the clinic and validation of target

Project Request: Animals:

Humans:

Clinical trial:

X

X

Patent number: PCT IT 2019 050073 Patent owner: IFO - IRCCS Istituto Regina Xxxxx 34% - IRCCS

Istituto Nazionale Tumori ¿Fondazione X. Xxxxxxx¿ 33% - La Sapienza Università di Roma 33%;

Project total financing request to the MOH: € 960.000

Free keywords: metastatic melanoma/BRAF/targeted therapy/drug resistance/microRNAs/lipid nanoparticles/delivery

Declarations

In case of a Synergy grant application 'Principal Investigator'(PI) means 'corresponding Principal Investigator on behalf of all Principal Investigators', and 'Host Institution' means 'corresponding Host Institution'.

1) The Principal Investigator declares to have the written consent of all participants on their participation and on the content of this proposal, as well as of any researcher mentioned in the proposal as participating in the project (either as other PI, team member or collaborator).	X
2) The Principal Investigator declares that the information contained in this proposal is correct and complete.	X
3) The Principal Investigator declares that all parts of this proposal comply with ethical principles (including the highest standards of research integrity — as set out, for instance, in the European Code of Conduct for Research Integrity — and including, in particular, avoiding fabrication, falsification, plagiarism or other research misconduct).	X
4) The Principal Investigator is only responsible for the correctness of the information relating to his/her own organisation. Each applicant remains responsible for the correctness of the information related to him and declared above.	X

Personal data protection

The assessment of your grant application will involve the collection and processing of personal data (such as your name, address and CV), which will be performed pursuant to Regulation (EC) No 45/2001 on the protection of individuals with regard to the processing of personal data by the Community institutions and bodies and on the free movement of such data. Unless indicated otherwise, your replies to the questions in this form and any personal data requested are required to assess your grant application in accordance with the specifications of the call for proposals and will be processed solely for that purpose. Details concerning the purposes and means of the processing of your personal data as well as information on how to exercise your rights are available in the privacy statement. Applicants may lodge a complaint about the processing of their personal data with the European Data Protection Supervisor at any time.

Abstract

BRAF-mutated metastatic melanoma relapsing after targeted or immunotherapy is an aggressive disease with high unmet clinical need. Only available therapeutic option is standard chemotherapy with dacarbazine with limited life expectancy.

Three members of this team (UO1, UO2, UO4) have recently identified and patented the use of a subset of microRNAs (miRNAs) in combination with targeted therapy to revert drug resistance (PCT/IT2019/050073: miRNAs for treatment and diagnosis of drug resistant tumors). The claims of this patent include systemic delivery of oncosuppressor miRNAs using lipid nanoparticles (LNPs). Our joint teams have generated in vitro and in vivo data demonstrating the antitumor efficacy of xxXXX-loaded LNPs (LNP-miRs) in combination with targeted therapy in melanoma models. The antitumor efficacy of LNP- miRs is explained by their capability of simultaneously hitting many intrinsic and extrinsic oncogenic avenues adopted by melanoma cells to survive upon MAPK pathway inhibition.

Inspired by these previous achievements, this project aims at optimizing LNP-miRs composition and completing pre-clinical efficacy and preliminary toxicology in preparation to clinical translation into Phase I-II studies.

The activities planned in order to advance to the future clinical trial will be structured into three highly focused experimental tasks. In particular, we plan to: 1) define a final, reproducible, easy to formulate and easily scalable production of oncosuppressor miRNAs loaded LNPs (LNP-miRs); 2) execute preliminary toxicology studies demonstrating optimal biodistribution, elimination and tolerability of LNP-miRs; 3) demonstrate therapeutic efficacy of these LNP-miRs in a variety of drug resistant human melanoma in vivo models in mice, starting from melanoma xenografts of drug resistant stable cell lines up to the final validation in PDX mice derived from tumour lesions of drug resistant patients.

We believe that the completion of these efforts will be possible in a two-year time frame and will represent a strong proof-of- concept of an innovative therapeutic paradigm whose main innovation consists in the use of a novel RNA therapeutic. The translational relevance of the project for public health, will be the definition of a new combination therapy for metastatic melanoma patients who have no other reliable therapeutic options available. The market value of our approach will be enhanced by the wealth of pre-clinical data we plan to collect and will significantly increase the probability to obtain additional financial support to conduct a Phase I/II clinical trial in the following years.

Yes

In order to best review your application, do you agree that the above non-confidential proposal title and abstract can be used, without disclosing your identity, when contacting potential reviewers?

2 - Participants & contacts

Operative Units
Institution that perform as UO	CF Institution	Department / Division / Laboratory	Role in the project	Southern Italy	SSN
1 - Istituti fisioterapici ospitalieri - Istituto Regina Xxxxx	02153140583	Scientific Direction - Preclinical Models and New Therapeutic Agents	Coordination		X
2 - Sapienza - University of Rome	80209930587	Department of Clinical & Molecular Medicine	Pathological Anatomy analysis
3 - Università degli Studi di Xxxxxx Xxxxxxxx XX	00876220633	Department of Pharamcy	Development of lipid nanoparticles encapsulating miRNAs	X
4 - IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx	00911350635	Dipartimento di Ricerca Traslazionale a supporto dei percorsi oncologici/Experimental Pharmacology Unit/ Mercogliano Systems Biology and Bioinformatics Laboratory	Collaboration to PDX models development, data analysis and interpretation; Recruitment, staging and treatment of eligible patients	X	X

Principal Research Collaborators
Key Personnel Name	Operative Unit	Role in the project
1 - del bufalo donatella	Istituti fisioterapici ospitalieri - Istituto Regina Elena	In vitro and in vivo preclinical studies with LNP- miRs formulations
2 - Xxxxxxx Xxxx	Xxxxxxxx - University of Rome	Contribution to in vitro assays and to in vivo pre-clinical studies. Pathology analysis of tumors from treated mice
3 - De Xxxx Xxxxxxxx	Università degli Studi di Xxxxxx Xxxxxxxx XX	Development of lipid nanoparticles encapsulating miRNAs
4 - Xxxxxxxx Xxxxx Xxxxxxx	IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx	Recruitment, staging and treatment of eligible patients. Coordination of samples collection, data analysis and interpretation.
5 - Xxxxxxxxxx Xxxxx	IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx	Collaboration to PDX models development, data analysis and interpretation
6 Under 40 - XXXXXXX XXXXX	Xxxxxxxx fisioterapici ospitalieri - Istituto Regina Xxxxx	Contribution to studies of in vitro cell lines and in vivo mouse models of drug resistance
7 Under 40 - DI XXXXXXX XXXXX	Xxxxxxxx fisioterapici ospitalieri - Istituto Regina Elena	In vitro and in vivo preclinical studies with LNP.miRs formulations

Key Personnel Name	Co-PI	Resp. CE	Resp. Animal	Birth Date	Gender
1 - del bufalo donatella	X			07/05/1961	F
2 - Xxxxxxx Xxxx				21/03/1967	F
3 - De Xxxx Xxxxxxxx				13/03/1973	M
4 - Xxxxxxxx Xxxxx Xxxxxxx				08/11/1964	M
5 - Xxxxxxxxxx Xxxxx				09/12/1977	F
6 Under 40 - XXXXXXX XXXXX				12/06/1985	M
7 Under 40 - DI XXXXXXX XXXXX				02/11/1988	F

Person in charge for the animal experiment: XXXXXXXXX XXXXXXX

Additional research collaborators under 40 to hire
Key Personnel Name	Operative Unit	Birth Date	Gender	Role in the project	Degree	Actual Pos. and Inst.
0 - Xxxx Xxxxxxx	Università degli Studi di Xxxxxx Xxxxxxxx XX	03/04/1992	F	Development of lipid nanoparticles encapsulating miRNAs	PhD	Post-doc fellow - Fondazione Xxxxxxx Xxxxxxxx
1 - Xxxxxxxx Xxxxxxx	IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx	06/05/1984	F	Development of PDX models	PhD	Post-doc fellow - Istituto Nazionale Tumori-IRCCS- Fondazione Xxxxxxx

2.1 Administrative data of participating

Operative Unit Number 1:

Address: IRCCS Istituto Nazionale Tumori Regina Xxxxx, Xxx Xxxx Xxxxxxxx 00, 00000, Xxxx, Xxxxx

PEC: xxxxxxxx@xxxx.xx

Operative Unit Number 2:

Address: Sapienza Università di Roma, Xxxxxxxx Xxxx Xxxx, 0, 00000 Xxxx, Xxxxx

PEC: xxxx.xxxxxxx@xxx.xxxxxxxx.xx

Operative Unit Number 3:

Address: Università degli Studi di Xxxxxx Xxxxxxxx XX, Xxx Xxxxxxxx Xxxxxxxxx, 00, 00000 Xxxxxx, Xxxxx

PEC: N/A

Operative Unit Number 4:

Address: IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx, Xxx Xxxxxxx Xxxxxxx, 00, 00000 Xxxxxx; Xxxxx

PEC: xxxxx.xxxxxxxx@xxx.xx

Operative Unit Number 5 (self financing):

Address: N/A

PEC: N/A

2.2 Principal Investigator (PI) Profile

Last Name: XXXXXXXXX

First Name: XXXXXXX

Last name at birth:

Gender: M

Title: Principal investigator

Nationality: Italiana

Date of birth: 22/09/1954

Official H index (Scopus or Web of Science): 70.0

Country of residence: ITALY Country of Birth: ITALY Place of Birth: Napoli

Scopus Author Id:55368402800 ORCID ID:0000-0003-2851-8605 RESEARCH ID:J-4131-2017

Contact address

Current organisation name: Istituti fisioterapici ospitalieri - Istituto Regina Elena

Current Department / Faculty / Institute / Laboratory name: Scientific Direction - Preclinical Models and New Therapeutic

Agents

Street: Xxx Xxxx Xxxxxxxx 00

Postcode / Cedex: 00144 Town: Roma

Phone:x000000000000 Phone 2: x00 00 0000 0000

Education / training
Educational institution and location	Degree	Field of study	From year	To year
European Molecular Biology Laboratory	PhD	Transcriptional control of RNA Polymerase III genes	1983	1983
Università degli Studi di Napoli "Xxxxxxxx XX", Naples Italy	PhD	Biochemistry of post- transcriptional methylation of transfer DNA	1979	1979
Università degli Studi di Napoli "Xxxxxxxx XX", Naples Italy	Master's Degree / Laurea Magistrale	Specialising in Biochemistry of post transcriptional pseudourydilation of trasfer DNA	1978	1978

Personal Statement:

Xxxx. Xxxxxxx Xxxxxxxxx is a molecular and cellular biologist with expertise in the study of mechanisims of drug resistance in cancer, signal transduction by cytokines and growth factors, cancer cell biology and genetics, immunology and immunotherapy of cancer. He has more than 30 years of research management experience as a coordinator of several research projects. In particular, in the last years he has worked in close collaboration with Xxxx. X. Xxxxxxx at Sapienza University of Rome on the study of lung cancer stem cells using as a source malignant pleaural effusions, more recently focusing on their interaction with elements of the tumour microenvironment which is the topic of this proposal. In this project Xxxx. Xxxxxxxxx will act as coordinator and supervisor of all activities.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
IRCCS Istituto Nazionale dei Tumori "Regina Elena"	Scientific Direction	Rome, Italy	Scientific Director	2016	2022
Istituto Nazionale dei Tumori IRCCS "Fondazione Xxxxxxx"	Scientific Direction	Naples, Italy	Scientific Director	2012	2016
Università degli Studi di Catanzaro "Magna Graecia"	Department of Molecular Biology & Cellular Biology	Catanzaro, Italy	Full Professor of Molecular Biology & Cellular Biology	0000	0000
IRBM- Istituto di Ricerche di Biologia Molecolare X.Xxxxxxxxx	Scientific Direction	Pomezia, Italy	Scientific Director	2006	2009
IRBM- Istituto di Ricerche di Biologia Molecolare X.Xxxxxxxxx	Department of Molecular & Cellular Biology	Pomezia, Italy	Head of Molecular & Cellular Biology	0000	0000
IRBM- Istituto di Ricerche di Biologia Molecolare X.Xxxxxxxxx	Department of Genetics	Pomezia, Italy	Head of the Genetics Department	1991	2001
Università degli Studi di Napoli "Xxxxxxxx XX"	Faculty of Molecular Biology	Naples, Italy	Full Professore of Molecular Biology	1990	1991
Università degli Studi di Pisa	Department of Biology	Pisa, Italy	Associate Professor of Genetics	1987	1990
Consiglio Nazionale delle Ricerche	Ricerca	Naples, Italy	Ricercatore	1985	1987
European Molecular Biology Laboratory	Department of Molecular Biology	Heidleberg, Germany	Staff Scientific	1983	1985

Other awards and honors

Xxxx. Xxxxxxx Xxxxxxxxx was elected a member Of EMBO in year 1998

From 2012-2016 was nominated Component of the Italian national Committee of Biosafety, Biotechnology and Life Sciences of the Presidenza dei Ministri

Other CV informations

-Since 2014 to present has been member of the Executive committee of the Alliance Against Cancer (ACC)

-Member of several national (SIBBM, SIC, FISV) and international (SITC, AAAS, AACR, EACR) scientific societies

-Starting from January 2017 President of FISV (Italian Federation of Scientiific Societies of Life Sciences)

-Since April 21 President of DIGICORE (Digital Institute for Cancer outcomes Research)

-Extensive experience in Technology Transfer as witnessed by Co-inventorship in about 30 international granted patents/patent applications

-Reviewer and editorial board member of several international scientific journals (CDDIS, JTRM, JITC, Cancers) and editor of textbook for graduate students

Selected peer-reviewed publications of the PI valid for minimum expertise level
Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**	P.*
Multi-omic approach identifies a transcriptional network coupling innate immune response to proliferation in the blood of COVID-19 cancer patients	Article	1019	12	2021	10.1038/s41419-021- 04299-y	34716309	1	L

Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**	P.*
Drug repurposing against COVID-19: focus on anticancer agents	Review	86	39	2020	10.1186/s13046-020- 01590-2	32398164	35	F
The emerging role of cancer cell plasticity and c ell-cycle quiescence in immune escape	Note	000	00	0000	10.1038/s41419-020- 2669-8	32555172	8	C
Boosting the arsenal against COVID-19 through computational drug repurposing	Editorial	946-948	25	2020	10.1016/x.xxxxxx.2020.04 .005	32304645	31	F
Reprogramming miRNAs global expression orchestrates development of drug resistance in BRAF mutated melanoma	Article	1267- 1282	26	2019	10.1038/s41418-018- 0000-0	00000000	00	L
Single cell analysis to dissect molecular heterogeneity and disease evolution in metastatic melanoma	Review	000	00	0000	10.1038/s41419-019- 2048-5	31672982	24	L
ErbB3 Phosphorylation as Central Event in Adaptive Resistance to Targeted Therapy in Metastatic Melanoma: Early Detection in CTCs during Therapy and Insights into Regulation by Autocrine Neuregulin	Article	1425	11	2019	10.3390/cancers111014 25	31557826	14	L
Combinations of immuno-checkpoint inhibitors predictive biomarkers only marginally improve their individual accuracy	Article	000	00	0000	10.1186/s12967-019- 1865-8	31014354	13	L
Metabolic features of cancer stem cells: the emerging role of lipid metabolism	Review	2367- 2378	37	2018	10.1038/s41388-018- 0141-3	29445137	63	L
Stearoyl-CoA-desaturase 1 regulates lung cancer stemness via stabilization and nuclear localization of YAP/TAZ	Article	4573- 4584	36	2017	10.1038/onc.2017.75	28368399	79	C
MicroRNAs in melanoma development and resistance to target therapy	Review	22262- 22278	8	2017	10.18632/oncotarget.147 63	28118716	71	L
Ferritin heavy chain is a negative regulator of ovarian cancer stem cell expansion and epithelial to mesenchymal transition	Article	62019- 62033	7	2016	10.18632/oncotarget.114 95	27566559	41	C
miR-579-3p controls melanoma progression and resistance to target therapy	Article	E5005-13	113	2016	10.1073/pnas.16077531 13	27503895	71	L
Stearoyl-CoA desaturase-1 is a key factor for lung cancer-initiating cells	Article	e947	4	2013	10.1038/cddis.2013.444	24309934	91	C
Activation of an early feedback survival loop involving phospho-ErbB3 is a general response of melanoma cells to RAF/MEK inhibition and is abrogated by anti-ErbB3 antibodies	Article	000	00	0000	10.1186/1479-5876-11- 180	23890105	52	L

* Position: F=First L=Last C=Correspondent O=Other N=Not applicable

** Autocertificated

Selected peer-reviewed publications of the PI for the evaluation CV
Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**
International validation of the consensus Immunoscore for the classification of colon cancer:	Article	2128- 2139	391 (00000)	0000	00.0000/S0140- 0000(00)00000-X	00000000	864
a prognostic and accuracy study

Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**
Abscopal effects of radiotherapy on advanced melanoma patients who progressed after ipilimumab immunotherapy	Article	e28780	0 (0)	0000	00.0000/onci.28780		256
Immunological and biological changes during ipilimumab treatment and their potential correlation with clinical response and survival in patients with advanced melanoma	Article	675-683	63 (0)	0000	00.0000/s00262-014- 1545-8	24695951	194
Quantitative optical imaging of primary tumor organoid metabolism predicts drug response in breast cancer	Article	5184- 5194	74 (00)	0000	00.0000/0008- 5472.CAN-14-0663	25100563	179
Triple negative breast cancer: Looking for the missing link between biology and treatments	Article	26560- 26574	6 (00)	0000	00.00000/oncotarget.530 6	26387133	112
Immunoscore and Immunoprofiling in cancer: An update from the melanoma and immunotherapy bridge 2015	Article	000	00 (0)	0000	00.0000/s12967-016- 1029-z	27650038	96
The additional facet of immunoscore: Immunoprofiling as a possible predictive tool for cancer treatment	Article	00	00 (0)	0000	00.0000/1479-5876-11- 54	23452415	92
Stearoyl-CoA desaturase-1 is a key factor for lung cancer-initiating cells	Article	e947	0 (00)	0000	00.0000/cddis.2013.444	24309934	91
Tumor genotype and immune microenvironment in POLE-ultramutated and MSI-hypermutated Endometrial Cancers: New candidates for checkpoint blockade immunotherapy?	Article	61-68	48	2016	10.1016/j.ctrv.2016.06.0 08	27362548	81
Stearoyl-CoA-desaturase 1 regulates lung cancer stemness via stabilization and nuclear localization of YAP/TAZ	Article	4573- 4584	36 (32)	2017	DOI: 10.1038/onc.2017.75	28368399	78

** Autocertificated

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
Istituto Italiano	IRCCS Istituto Nazionale Tumori	2021	Sviluppo di approcci e	Coordinator	142.000,00	xxxxx://xxxxxxxxxxxx.xx
Buddista Soka Gakkai	Regina Elena		metrica per valutare			xxxxxxxx.xx/xxxxxxxx/x
			l¿impatto e il miglioramento			ometa/
			della prognosi in pazienti
			con fragilità nell¿epoca
			Covid-19 - COMETA
AIRC	IRCCS Istituto Nazionale Tumori	2018 -	A microRNA-based	Coordinator	730.000,00	xxxxx://xxx.xxxx.xx/xxx
	Xxxxxx Xxxxx	2022	approach to advanced			ercatori/i-nostri-
			diagnosis and therapy of			ricercatori/xxxxxxx-
			metastatic melanoma			xxxxxxxxx

Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
IMI Intergruppo Melanoma Italiano	IRCCS Istituto Nazionale Tumori Regina Xxxxx	2018	A xxXXX-based signature as predictive biomarker of drug resistance in melanoma	Coordinator	30.000,00	xxxxx://xxx.xxxxxx xxxxx.xx/xxxxxxxxxxx/ article/639/82%20Ve rbale%20Pubblica% 20Selezione%20per %20il%20Finanziam ento%20di%20n.%2 02%20Progetti%20S cientifici.pdf
MUR PROSCAN	IRCCS Istituto Nazionale Tumori Regina Xxxxx	2018	Strumenti micro-meccanici e robotici per la diagnosi e la terapia del cancro alla prostate	Collaborator	141.717,32	xxxxx://xxx.xxxx.xxx .it/documents/20182/ 369540/DD+n.+2013 +Scheda+Costi.pdf/1 xx0x000-x0x0-000x- x0x0- x000x0xxxx0x?xxxxx on=1.0&t=15379746 21659
AIRC	IRCCS Istituto Nazionale Tumori Fondazione X Xxxxxxx	2015 - 2017	ERBB3 Receptor as a new key player in melanoma progression	Coordinator	330.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/xxxxxxx- xxxxxxxxx
AIRC	Università degli Studi di Catanzaro "Magna Graecia"	2011 - 2013	Targeting the HEDR3 receptor with monoclonal antibodies	Coordinator	255.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/xxxxxxx- xxxxxxxxx

2.3 CO-PI Profile

Last Name: del bufalo

First Name: donatella

Last name at birth:

Gender: F

Title: In vitro and in vivo preclinical studies with LNP-miRs formulations

Nationality: italiana

Date of birth: 07/05/1961

Official H index (Scopus or Web of Science): 35.0

Country of residence: ITALY

Country of Birth: ITALY

Place of Birth: Montopoli di Xxxxxx

Xxxxxx Author Id:6701629065 ORCID ID:0000-0002-3148-6096 RESEARCH ID:K-8673-2016

Contact address

Current organisation name: Istituti fisioterapici ospitalieri - Istituto Regina Elena

Current Department / Faculty / Institute / Laboratory name: Scientific Direction - Preclinical Models and New Therapeutic

Agents

Street: xxx Xxxx Xxxxxxxx 00

Postcode / Cedex: 00144 Town: Roma

Phone:x000000000000 Phone 2: 0000000000

Education / training
Educational institution and location	Degree	Field of study	From year	To year
Sapienza University, Rome, Italy	Specialization / Specializzazione	Role of Bcl-2 in tumor metastatizzation	1987	1991
Sapienza University, Rome, Italy	Single-cycle master's degree / Laurea magistrale a ciclo unico	Analysis of factors involved in the formation of chromatid aberrations in the G2 phase of the cell cycle: study with different inhibitors	1980	1985

Personal Statement:

Xxx Xxxxxx'x research is focused on basic as well as on translational oncology, devoted to study in vitro and in vivo the role of different genetic pathways in cancer progression and response to therapy, with particular regards to melanoma. Her publications confirm experience in the field of the proposal. As co-PI, together with the PI, she will i) monitor the proper advancement of the project controlling the connection between work plan and results obtained, ii) be in charge of the in vitro and in vivo preclinical studies to characterize and determine the basal toxicity of LNP formulations, and to evaluate the efficacy of LNP-miR in combination with targeted therapies. In collaborations with XX and other collaborators, co-PI will write scientific papers.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
IRCCS Regina Elena National Cancer Institute	Research and Advanced Technologies Department - Preclinical Models and New Therapeutic Agents Unit	Rome, Italy	Research Scientist, permanent position	2016	2022
IRCCS Regina Elena National Cancer Institute	Laboratory of Experimental Chemotherapy	Rome, Italy	Research Scientist, permanent position	1993	2014
Sapienza University	Faculty of Biological Science	Rome, Italy	Teaching activity	1992	1995
IRCCS Regina Elena National Cancer Institute	Laboratory of Experimental Chemotherapy	Rome, Italy	Junior Research Scientist	1992	1992
NCI-National Cancer Institute Cancer Institute	Molecular and Cellular Immunology Laboratory	Frederick, MD, USA	Visiting Scientist	1990	1991
IRCCS Regina Elena National Cancer Institute	Laboratory of Experimental Chemotherapy	Rome, Italy	Research Fellow	1987	1989

Other awards and honors

2021-present Executive committee (EM) BioBank IFO 2021-present EM Animal welfare IFO

2018-present EM Animal welfare Sapienza Univerity of Rome 2014-present Editorial Board some Journals

2012-16 EM European Association Cancer Research

2010-present Responsible for the use of animals for scientific purposes (Authorization by Italian Ministry of the Health) 2006-09 EM Italian Cancer Society

2000-04 EM AIRC Study Section

Other CV informations

Previously experience as PI of different funded grants (AIRC, Italian Ministry of Health, Research National Council, Fondazione Veronesi, Fondazione Calabresi), and as tutor of fellows, undergraduate and PhD students proved her skills for project planning, execution and dissemination. The strong expertise in cellular and molecular biology and in in vivo studies, documented by several authorizations from Italian Ministry of Health for animal experimentation (342/2019-PR, 563/2021), in combination with expertise from collaborative interdisciplinary projects and established external collaboration, provides a basis for successful project co-leadership.

Selected peer-reviewed publications of the Co-PI valid for minimum expertise level
Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**	P.*
Targeting the anti-apoptotic Bcl-2 family proteins: Machine learning virtual screening and biological evaluation of new small molecules	Article	2427- 2444	12	2022	10.7150/thno.64233	35265218	0	L
Bcl-xl: A focus on melanoma pathobiology	Review	1-17	22	2021	10.3390/ijms22052777	33803452	0	L
Antitumor effect of Melaleuca alternifolia essential oil and its main component terpinen-4- ol in combination with target therapy in melanoma models	Article	NOT_FO UND	7	2021	10.1038/s41420-021- 00510-3	34059622	6	L

Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**	P.*
New insights into the roles of antiapoptotic members of the Bcl-2 family in melanoma progression and therapy	Review	1126- 1135	26	2021	10.1016/x.xxxxxx.2021.01 .027	33545382	3	L
Essential oils and their main chemical components: The past 20 years of preclinical studies in Melanoma	Review	1-45	12	2020	10.3390/cancers120926 50	32948083	9	L
Inhibition of Anti-Apoptotic Bcl-2 Proteins in Preclinical and Clinical Studies: Current Overview in Cancer	Review	NOT_FO UND	9	2020	10.3390/cells9051287	32455818	31	L
Melanoma-specific bcl-2 promotes a protumoral M2-like phenotype by tumor-associated macrophages	Article	NOT_FO UND	8	2020	10.1136/jitc-2019- 000489	32269145	11	L
microRNA-378a-5p iS a novel positive regulator of melanoma progression	Article	NOT_FO UND	9	2020	10.1038/s41389-020- 0203-6	32060259	15	L
Targeting hypoxia in tumor: A new promising therapeutic strategy	Review	NOT_FO UND	39	2020	10.1186/s13046-019- 1517-0	31924239	14	L
Emerging role of histone acetyltransferase in stem cells and cancer	Review	NOT_FO UND	2018	2018	10.1155/2018/8908751	30651738	30	L
Semaphorin 5A drives melanoma progression: Role of Bcl-2, miR-204 and c-Myb	Article	NOT_FO UND	37	2018	10.1186/s13046-018- 0933-x	30454024	14	L
Histone deacetylase inhibitor ITF2357 leads to apoptosis and enhances doxorubicin cytotoxicity in preclinical models of human sarcoma	Article	NOT_FO UND	7	2018	10.1038/s41389-018- 0026-x	29472530	12	L
BCL-X<inf>L</inf> overexpression promotes tumor progression-associated properties article	Article	NOT_FO UND	8	2017	10.1038/s41419-017- 0055-y	29238043	46	L
Non-canonical roles of Bcl-2 and Bcl-xL proteins: Relevance of BH4 domain	Review	579-587	38	2017	10.1093/carcin/bgx016	28203756	27	L
Affinity purification-mass spectrometry analysis of bcl-2 interactome identified SLIRP as a novel interacting protein	Article	NOT_FO UND	7	2016	10.1038/cddis.2015.357	26866271	8	L
miR-211 and MITF modulation by Bcl-2 protein in melanoma cells	Article	2304- 2312	55	2016	10.1002/mc.22437	26599548	17	L
Kinetochore-microtube attachments in cancer therapy	Editorial	902-903	2	2015	10.18632/oncoscience.2 65	26697517	0	F
Histone deacetylase inhibition synergistically enhances pemetrexed cytotoxicity through induction of apoptosis and autophagy in non- small cell lung cancer	Article	NOT_FO UND	13	2014	10.1186/1476-4598-13- 230	25301686	44	F
BH4 domain of bcl-2 protein is required for its proangiogenic function under hypoxic condition	Article	2558- 2567	34	2013	10.1093/carcin/bgt242	23836782	21	L
Removal of the BH4 domain from Bcl-2 protein triggers an autophagic process that impairs tumor growth	Article	315-327	15	2013	10.1593/neo.121392	23479509	26	L

* Position: F=First L=Last C=Correspondent O=Other N=Not applicable

** Autocertificated

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
AIRC	IRCCS Regina Elena National Cancer Institute	2021 - 2025	Shaping melanoma microenvironment by bcl-2: from novel mediators of tumor/stroma crosstalk to new therapeutic approaches	Coordinator	750.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/donatella- del-bufalo
Banca d'Italia	IRCCS Regina Elena National Cancer Institute	2022	Identificazioni di nuove strategie terapeutiche per il trattamento del melanoma	Coordinator	25.000,00	xxxxx://xxx.xxxxxxxx xxxx.xx/xxx- siamo/impegno- ambientale- sociale/index.html?d otcache=refresh
AIRC	IRCCS Regina Elena National Cancer Institute	2017 - 2019	Spanning bcl-2 functions in melanoma models: from microenvironment to microRNA modulation	Coordinator	450.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/donatella- del-bufalo
AIRC	IRCCS Regina Xxxxx National Cancer Institute	2014 - 2016	Digging deeper into regulation of autophagy and angiogenesis by bcl-2 family proteins	Coordinator	450.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/donatella- del-bufalo
AIRC	IRCCS National Cancer Institute Regina Elena	2011 - 2013	Cross-talk between bcl-2 and hypoxia: insight into molecular events leading to tumor progression	Coordinator	330.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/donatella- del-bufalo
AIRC	IRCCS National Cancer Institute Regina Xxxxx	2008 - 2010	Interplay between bcl-2 and hypoxia: implication for cancer therapy	Coordinator	240.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/donatella- del-bufalo
AIRC	IRCCS National Cancer Institute Regina Xxxxx	2005 - 2007	Studies of the mechanisms by which bcl-2 overexpression increases angiogenic activity	Coordinator	240.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/donatella- del-bufalo
Ministry of Health	IRCCS National Cancer Institute Regina Xxxxx	2003 - 2005	Identification and characterization of inhibitors of chemotactic factors for the treatment of neoplasia and inflammatory patologies	Collaborator	101.200,00	NA
AIRC	IRCCS National Cancer Institute Regina Xxxxx	2002 - 2004	Study of the mechanism by which bcl-2 modulates angiogenesis	Coordinator	78.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/donatella- del-bufalo
Ministry of Health	IRCCS National Cancer Institute Regina Xxxxx	2001 - 2003	Studies of the molecular mechanisms by which bcl-2 modulates the response to antineoplastic treatments	Coordinator	33.000,00	NA

2.3 Research Collaborators n. 2

Last Name: Xxxxxxx

First Name: Xxxx

Last name at birth:

Gender: F

Title: Contribution to in vitro assays and to in vivo pre-clinical studies. Pathology analysis of tumors from treated mice

Nationality: italiana

Date of birth: 21/03/1967

Official H index (Scopus or Web of Science): 27.0

Country of residence: ITALY Country of Birth: ITALY Place of Birth: Isola del Liri

Scopus Author Id:7102285810 ORCID ID:0000-0002-5491-2449 RESEARCH ID:K-8493-2016

Contact address

Current organisation name: Sapienza - University of Rome

Current Department / Faculty / Institute / Laboratory name: Department of Clinical & Molecular Medicine

Street: via di grottarossa 1035/1039

Postcode / Cedex: 00189 Town: roma

Phone:x000000000000 Phone 2:

Education / training
Educational institution and location	Degree	Field of study	From year	To year
Sapienza University of Rome, Rome, Italy	Specialization / Specializzazione	Establishment of a bio-bank of MPE-derived stem cells.Analysis of CSC derived from Malignant Pleural Effusion (MPE) of patients affected by lung adenocarcinoma.	2004	2008
Sapienza University of Rome, Rome, Italy	PhD	generation and characterization of primary cultures of lung adenocarcinoma; propagation of CSC derived from Malignant Pleural Effusion (MPE) of patients affected by lung adenocarcinoma	1994	1999
Sapienza University of Rome, Rome, Italy	Single-cycle master's degree / Laurea magistrale a ciclo unico	Cangerogenesis in cervical and breast cancer.	1989	1994

Personal Statement:

Xxxx. Xxxx Xxxxxxx has extensive experience in the study of non small cell lung cancer (NSCLC) and in the isolation and characterization of primary tumour cells from MPEs of patients with NSCLC. In addition, in recent years she has dedicated herself to the study of the tumour microenvironment by isolating PBMC from peripheral blood of patients with lung adenocarcinoma and PEMC from the pleural effusions collected from the same patients. Her crucial role in the project will be to coordinate the enrollment of patients and the biobanking of biological material. She will also be involved in the coordination of activities concerning the culturing of tumour and immune cells for the setting of ex vivo models necessary

for the validation of the identified potential targets.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
Sapienza University of Rome	Department of Clinical and Molecular Medicine	Roma, Italy	Full Professor	2021	2022
Sapienza University of Rome	Department of Clinical and Molecular Medicine	Rome, Italy	Assistant Professor	2019	2021
Sapienza University of Rome	Department of Clinical and Molecular Medicine	Rome, Italy	Researcher Clinical Pathology	2006	2019

Other awards and honors

2001-2003 Fellowship of the American Italian Cancer Foundation at Xxxxxx Cancer Centre, Xxxxxx Xxxxxxxxx University, Xxxxxxxxxxxx, XX, XXX

0000-0000 Fellowship of the American Italian Cancer Foundation at La Spaineza University of Rome

2018-present President in Laurea Magistrale in Technical and Diagnostic Health Professions (SSD06/N1-Med46)

2020-2023 Junior Research Fellow Superior School Advance Studies Sapienza (SSAS)-Academic Class "Life Sciences"

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
Istituto Italiano Buddista Soka Gakkai	Università degli Studi di Roma "Sapienza"	2021 - 2022	Sviluppo di approcci e metrica per valutare l'impatto e il miglioramento della prognosi in pazienti con fragilità nell'epoca Covid-19	Collaborator	99.000,00	xxxxx://xxxxxxxxxxxx.xx xxxxxxxx.xx/xxxxxxxx/x ometa/
Regione Lazio - Gruppi di Ricerca	Università degli Studi di Roma "Sapienza"	2021 - 2023	Deconvoluzione del Microambiente tumorale polmonare metastatico ed identificazione di nuovi bersagli per l'Immunoterapia	Coordinator	149.999,85	xxxxx://xxx.xxxxxxx.x xxxx.xx/xxxxx/xxxxxxx/xxx es/documentazione/ SVI_RIC_G04014_13 _04_2021_Allegato2. pdf
AIRC	Università degli Studi di Roma "Sapienza"	2020 - 2025	Cellule staminali tumorali e metabolismo lipidico: capire il ruolo di SCD1 come bersaglio terapeutico e biomarker predittivo	Coordinator	456.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/xxxx- xxxxxxx
Università degli Studi di Roma "Sapienza"	Università degli Studi di Roma "Sapienza"	2019	Biobanks of primary cultures enriched with Lung Cancer Stem Cells (CSC) and adapted immune cells: targeting lipid metabolism and correlations patients risk factors	Coordinator	29.000,00	xxxxx://xxx.xxxxxxx0 .it/dip_dmcm/node/66 37

Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
PRIN	Università degli Studi di Roma "Sapienza"	2019 - 2021	Ruolo di YAP/TAZ sui regolatori di trascrizione nel cancro chemio-resistente	Collaborator	98.547,00	xxxxx://xxxx.xxx.xxx.xx /Ricerca?Filtro.Anno =%25&Filtro.Ateneo= Universit%C3%A0+d egli+Studi+di+ROMA +%22La+Sapienza% 22&Filtro.Argomento =YAP%2FTAZ&Filtro .Cognome=
Lazio Innova	Università degli Studi di Roma "Sapienza"	2018 - 2021	Cellule staminali neoplastiche e immuno- evasione nell'adenocarcinoma polmonare	Coordinator	138.000,00	xxxxx://xxx.xxxxxxxxx xx.xx/xxx/xxxxxxx/00 18/06/BUR-2018-53- 0-252-278- estratto.pdf
AIRC	Università di Roma "Sapienza"	2015 - 2018	Studio su Stearoyl CoA Desaturasi 1 (SCD1) come fattore chiave di inizio delle cellule tumorali di tumore del polmone non a piccole	Coordinator	357.000,00	xxxxx://xxx.xxxx.xx/xxx ercatori/i-nostri- ricercatori/xxxx- xxxxxxx

2.4 Research Collaborators n. 3

Last Name: Xx Xxxx

First Name: Xxxxxxxx

Last name at birth:

Gender: M

Title: Development of lipid nanoparticles encapsulating miRNAs

Nationality: Italiana

Date of birth: 13/03/1973

Official H index (Scopus or Web of Science): 34.0

Country of residence: ITALY Country of Birth: ITALY Place of Birth: Napoli

Scopus Author Id:7102090682 ORCID ID:0000-0002-1210-1269 RESEARCH ID:G-6343-2012

Contact address

Current organisation name: Università degli Studi di Xxxxxx Xxxxxxxx XX

Current Department / Faculty / Institute / Laboratory name: Department of Pharamcy

Street: Xxx Xxxxxxxx Xxxxxxxxx 00

Postcode / Cedex: 80131 Town: Napoli

Phone:x000000000000 Phone 2:

Education / training
Educational institution and location	Degree	Field of study	From year	To year
Università degli Studi di Cagliari - Cagliari (in Consortium with the Università degli Studi di Xxxxxx Xxxxxxxx XX - Naples).	PhD	Drug delivery of proteins and nucleic aicids	1998	2001
Degree in Pharmaceutical Chemistry and Technology	Single-cycle master's degree / Laurea magistrale a ciclo unico	Pharmaceutical Chemistry and Technology	1991	1997

Personal Statement:

The scientific activity can be summarized as the design and development of drug delivery systems based on micro- and nanotechnologies, especially focused on the development of liposomes and other lipid-based nanovectors. Prof. Xx Xxxx is author of about 100 papers (H-index: 34) on international journals, 4 book chapters and different patent applications (with granted PCT, US and EU patent). Dr Xx Xxxx is member of the Editorial Board of different journals in the field of drug delivery and pharmaceutics. Dr Xx Xxxx is a Board member of ADRITELF and Divisione di Tecnologia Farmaceutica SCI, and in the past of the Controlled Release Society (CRS) Italy Chapter.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
Università degli Studi di Xxxxxx Xxxxxxxx XX	Drug Delivery Group - BioNanoMed	Xxx Xxxxxxxx Xxxxxxxxx 00, Xxxxxx (Xxxxx)	Full professor	2021	2022
Università degli Studi di Xxxxxx Xxxxxxxx XX	Drug Delivery Group - BioNanoMed	Xxx Xxxxxxxx Xxxxxxxxx 00, Xxxxxx (Xxxxx)	Associate professor	2014	2021
Università degli Studi di Xxxxxx Xxxxxxxx XX	Drug Delivery Group - BioNanoMed	Xxx Xxxxxxxx Xxxxxxxxx 00, Xxxxxx (Xxxxx)	Assistant professor	2002	2014

Other awards and honors

Premio ADRITELF 2020 consegnato nel xxxxx xxxxx "00xx World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology", maggio 2021

Premio Speciale per il poster presentato nell'ambito del 53° Simposio AFI che si è tenuto a Rimini nei giorni 12-14 giugno 2013.

Premio nell'ambito di Campani Start Cup 2011 per il progetto NANOPHARM.

Premio Speciale per il poster presentato nell'ambito del 51° Simposio AFI che si è tenuto a Rimini nei giorni 8-10 giugno 2011.

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
Phospholipid Research	Università di Xxxxxx Xxxxxxxx XX	2021	Engineering Lipid Self-	Coordinator	46.000,00	xxxxx://xxx.xxxxxxxx
Center Funding			Assembling Nanoparticles			ipid-research-
			as Novel mRNA-based			xxxxxx.xxx/xxxxxxx/x
			Vaccine Platforms			ngineering-lipid-self-
						assembling-
						nanoparticles-as-
						novel-mrna-based-
						vaccine-platforms/
Phospholipid Research	Università degli Studi di Napoli	2020	Lipid nanovectors to use	Coordinator	30.000,00	xxxxx://xxx.xxxxxxxx
Center Funding	Federico II		non-coding RNA			ipid-research-
			oligonucleotides in			xxxxxx.xxx/xxxxxxx/xxx
			glioblastoma in combination			id-nanovectors-to-
			with standard therapy -Part			use-non-coding-rna-
			2			oligonucleotides-in-
						glioblastoma-in-
						combination-with-
						standard-therapy-
						part-2/

Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
Phospholipid Research Center Funding	Università degli Studi di Xxxxxx Xxxxxxxx XX	2018	Lipid nanovectors to use non-coding RNA oligonucleotides in glioblastoma in combination with standard therapy	Coordinator	35.000,00	xxxxx://xxx.xxxxxxxx ipid-research- xxxxxx.xxx/xxxxxxx/xxx id-nanovectors-to- use-non-coding-rna- oligonucleotides-in- glioblastoma-in- combination-with- standard-therapy
BioCam PerMedNet	Università degli Studi di Xxxxxx Xxxxxxxx XX	2018	MEDICINA PERSONALIZZATA PER STRATEGIE INNOVATIVE IN MALATTIE NEUROPSICHIATRICHE E VASCOLARI - PerMedNet	Collaborator	8.954.886,00	xxxxx://xxx.xxxxxx.x u/progetti-di- ricerca/permednet_p 3.html
Regione Campania - POR CAMPANIA FESR 2014/2020 - MANIFESTAZIONE DI INTERESSE PER LA REALIZZAZIONE DI TECHNOLOGY PLATFORM NELL'AMBITO DELLA LOTTA ALLE PATOLOGIE ONCOLOGICHE X.X. XX 00 10 N. 355 DEL 05/06/2017 E SS.MM.II.	Università degli Studi di Xxxxxx Xxxxxxxx XX	2018	Innovazioni diagnostiche e terapeutiche per tumori neuroendocrini, endocrini e per il glioblastoma attraverso una piattaforma tecnologica integrata di competenze cliniche, genomiche, ICT, farmacologiche e farmaceutiche	Collaborator	6.428.000,00	xxxxx://xxx.xxxx.xxxxx .it/handle/11588/787 014?mode=full.564
Ministero della Salute	"Università degli Studi di Xxxxxx Xxxxxxxx XX"	2007	Ricerca e sviluppo di nuovi gel bioattivi per la medicazione di piaghe da decubito (GR-2007-687941)	Coordinator	520.000,00	xxxx://xxx.xxxxxx.xxx .it
Università Xxxxx Xxxxxxxx	"Università degli Studi di Xxxxxx Xxxxxxxx XX"	2006	Liposomi cationici bioadesivi per la somministrazione oculare di oligonucleotidi antisenso	Coordinator	4.000,00	xxxxx://xxx.xxxxxxxxx e-franco- xxxxxxxxx.xxx/xxxx- principal/bandi/progr amma-galileo/
PRIN MUR	"Università degli Studi di Xxxxxx Xxxxxxxx XX"	2009	Sviluppo nanotecnologico di agenti citotossici e target- based nel trattamento dei tumori: nuove strategie basate su razionali biologici	Collaborator	442.725,00	xxxxx://xxxx.xxx.xxx.xx /Pages/Index/106

2.5 Research Collaborators n. 4

Last Name: Ascierto

First Name: Xxxxx Xxxxxxx

Last name at birth:

Gender: M

Title: Recruitment, staging and treatment of eligible patients.

Coordination of samples

collection, data analysis and interpretation.

Nationality: italiana

Date of birth: 08/11/1964

Official H index (Scopus or Web of Science): 84.0

Country of residence: ITALY Country of Birth: ITALY Place of Birth: SOLOPACA

Scopus Author Id:6701588348 ORCID ID:0000-0002-8322-475X RESEARCH ID:J-9106-2016

Contact address

Current organisation name: IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx

Current Department / Faculty / Institute / Laboratory name: Dipartimento di Ricerca Traslazionale a supporto dei percorsi

oncologici/Experimental Pharmacology Unit/ Mercogliano Systems Biology and Bioinformatics Laboratory

Street: XXX XXXXXXX XXXXXXX 00

Postcode / Cedex: 80131 Town: NAPOLI

Phone:x000000000000 Phone 2: 0000000000

Education / training
Educational institution and location	Degree	Field of study	From year	To year
Xxxxxxxxxx xxxxx Xxxxx xx Xxxxxx Xxxxxxxx XX, Xxxxxx (XX), Xxxxx	Specialization / Specializzazione	Oncology	1990	1994
Università degli Studi di Xxxxxx Xxxxxxxx XX, Napoli (NA), Italy	Single-cycle master's degree / Laurea magistrale a ciclo unico	Medicine and Surgery	1983	1990

Personal Statement:

Dr. Ascierto is a world leading key opinion leader in Immunotherapy. He is currently Director of the Department of Melanoma, Cancer Immunotherapy and Development Therapeutics at Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Xxxxxx, Italy. He is a Scientific Reviewer for leading medical journals including NEJM, JCO, Lancet

Oncology, & Clinical Cancer Research. He is a member of the Steering Committee of the Society of Melanoma Research, Board of Directors for SITC, & President of the Fondazione Melanoma Onlus, & Campania Society of ImmunoTherapy of Cancer. He has presided as PI on over 150 clinical trials, & he is the author of more than 550 publications in peer-reviewed journals, with a total IF of about 5500 and about 52000 citations.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Xxxxxx	Department of Melanoma, Cancer Immunotherapy and Development Therapeutics	Napoli, Italy	Director	2020	2022
Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	Unit of Melanoma, Cancer Immunotherapy and Innovative Therapy	Napoli, Italy	Director	2013	2022
Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	Unit of Melanoma, Cancer Immunotherapy and Innovative Therapy	Napoli, Italy	Vice-Director	2011	2013
Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	Unit of Medical Oncology and Innovative Therapy	Napoli, Italy	Director	2008	2011
Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Xxxxxx	Department of Clinical Immunology	Napoli, Italy	Vice-Director	2003	2008

Other awards and honors

From 2010 President of Melanoma Foundation From 2014 President of SCITO

From 2016 Member of the Board of SMR From 2017 Member of the Board of CDDF

From 2017 Member of Board of Directors for the SITC

2018 Award Zanibelli as Person of the Year in the fied of Heathcare and Cancer Research. 2018 Award Medicina Italia as excellence of 2018

2019 Award Xxxxx Xx Catena - Silver Medal for merit and Civic engagement 2019 Award Campania as Science Best Award

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
Ministero della Salute	Istituto Nazionale Tumori IRCCS	2022	PROGRAMMAZIONE	Coordinator	30.000,00	xxxxx://xxxxxxxxxxxx.x
	Fondazione X. Xxxxxxx Napoli		TRIENNALE RICERCA			xxx.xx/xxxxxxxxxxxx/x
			CORRENTE 2022-2024 -			reaprogetti
			MoH - WFR 2611811;
			Progetto dal Titolo
			"Valutazione dei
			meccanismi di resistenza e
			dei biomarcatori indicativi di
			risposta al trattamento con
			Immunoterapia e/o Farmaci
			Innovativi L2/1"

Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
ACC - Alleanza Contro il cancro	Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Xxxxxx	2021	Programma nazionale di oncologia personalizzata per gli IRCCS della Rete ACC -RCR-2021-2367121 - Progetto dal Titolo "ACC- ImmunoPortal, un data- repository italiano per l'ottimizzazione dell'immunoterapia con ICI: uno studio osservazionale ambispettico (WP6/WG6 Immunotherapy)"	Collaborator	14.737,00	xxxxx://xxx.xxxxxxxx xxxxxxxxxxxxxx.xx/xxxx etti/
POR Campania	Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	2020	POR FESR CAMPANIA 2014-2020 - CUP H64I20000300002; Titolo Progetto: "Caratterizzazione Bio-Molecolare del virus SARS-COV-2 e dei cofattori dell'infiammazione implicati nella patogenesi della COVID-19	Collaborator	799.999,26	xxxxx://xxxxxxx.xxxxxx x.xxxxxxxx.xx/xx/xxxx etti-e- beneficiari/progetti-e- beneficiari- 57ex/elenco-dei- progetti-e-dei- beneficiari?page=1
POR Regione Lombardia	Istituto Nazionale Tumori IRCCS Xxxxxxxxxx X. Xxxxxxx Xxxxxx	0000	Xxxxxxx Xxxxxxxxx POR FESR 2014/2020. Call HUB Ricerca ed Innovazione. CUP E61B19000600007; Titolo del Progetto "Sviluppo di nuove molecole di seconda generazione per immunoterapia oncologica (ImmunHUB)".	Collaborator	1.130.000,00	xxxxx://xxx.xxxx.xxxx xxx.xxxxxxxxx.xx/xxx/ portal/PROUE/FESR /beneficiari-e- operazioni- finanziate/elenco- beneficiari-e- operazioni/
AIRC	Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	2019	Metastatic disease: the key unmet need in oncology - Second Edition - Fondazione AIRC 5x1000 2018. - Id. 22757 - Progetto dal Titolo "Disease-specific universal vaccines as new combinatorial immunotherapy for metastatic melanoma, sarcoma and osteosarcoma - Acronimo UniCanVax".	Collaborator	156.860,00	xxxxx://xxxxxxxxx0xx xxxxxx.xxxx.xx/xxxxxxxxx azione/
ACC - Alleanza Contro il Cancro	Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	2019	Progetto di ricerca su cellule CAR-T per patologie ematologiche maligne e per tumori solidi (ACC Network) - CUP H64I19000770001	Collaborator	220.000,00	xxxxx://xxx.xxxxxxxx xxxxxxxxxxxxxx.xx/xxxx etti/car-t/
POR Campania	Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	2019	CUP B61C17000060007; Dalla Genomica alla Terapia dei Tumori Rari (Genomica e Terapia)	Collaborator	830.000,00	xxxxx://xxxxxxx.xxxxxx x.xxxxxxxx.xx/xx/xxxx etti-e- beneficiari/progetti-e- beneficiari- 57ex/elenco-dei- progetti-e-dei- beneficiari?page=1

Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
POR Campania	Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	2019	CUP B61G18000470007 "Combattere la resistenza tumorale: piattaforma integrata multidisciplinare per un approccio tecnologico innovativo alle oncoterapie (Campania Oncoterapie)"	Collaborator	3.100.000,00	xxxxx://xxxxxxx.xxxxxx x.xxxxxxxx.xx/xx/xxxx etti-e- beneficiari/progetti-e- beneficiari- 57ex/elenco-dei- progetti-e-dei- beneficiari?page=1
ERA NET TRANSCAN	Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	2014	ERA-NET on Translational Cancer Research (TRANSCAN) 2013 "Integrative cancer- immunology and immunoscore for cancer classification and immunotherapies (IMMUNOSCOR)"	Collaborator	242.000,00	xxxxx://xxx.xxxxxxxx xx0.xx/xxxxx.xxx/xxx tract/xxxxxxxxxx.xx ml
Ministero della Salute - Bando Ricerca Finalizzata	Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx Napoli	2011	CO-2011-02348049 - New Possible Approach for the Classification of Melanoma and Prediction of Response to Immunotherapy: Immunoscore and Immune Profiling"	Coordinator	150.000,00	xxxxx://xxxxxxxxxxxx.x xxx.xx/xxxxxxxxxxxx/x reaprogetti

2.6 Research Collaborators n. 5

Last Name: Xxxxxxxxxx

First Name: Xxxxx

Last name at birth:

Gender: F

Title: Collaboration to PDX models development, data analysis and interpretation

Nationality: Italiana

Date of birth: 09/12/1977

Official H index (Scopus or Web of Science): 32.0

Country of residence: ITALY

Country of Birth: ITALY

Place of Birth: San Paolo Bel Sito

Scopus Author Id:8320797100 ORCID ID:0000-0002-7538-4657 RESEARCH ID:J-9914-2018

Contact address

Current organisation name: IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx

Current Department / Faculty / Institute / Laboratory name: Dipartimento di Ricerca Traslazionale a supporto dei percorsi

oncologici/Experimental Pharmacology Unit/ Mercogliano Systems Biology and Bioinformatics Laboratory

Street: Istituto Nazionale Tumori - IRCCS - Fondazione X. Xxxxxxx - Laboratori di Mercogliano, via Ammiraglio Bianco

Postcode / Cedex: 83013 Town: Mercogliano

Phone:x000000000000 Phone 2: 0000000000

Education / training
Educational institution and location	Degree	Field of study	From year	To year
Xxxxxxxxxx Xxxxxxxx XX of Naples (Italy)	Specialization / Specializzazione	Specialty Certification in Clinical Pathology and Biochemistry	2013	2017
Second University of Naples, Naples	PhD	PhD in Computational Biology - Computational analysis to study the structure-function relationship in proteins	2000	2004
University of Naples "Federico II", Naples	Single-cycle master's degree / Laurea magistrale a ciclo unico	Degree in Chemistry-Application of NMR approach to study the structure of peptides	1995	2000

Personal Statement:

Dr. Xxxxxxxxxx'x research activity is focused on the study of cytokines and chemokines profiling as well as metabolomic/lipidomic analysis by 1H-Nuclear magnetic resonance (NMR) spectroscopy and Liquid chromatography-Mass Spectrometry, in various biological matrices of cancer patients. She has a long experience in the application of systems biology approaches to integrate omics and clinical data, and in the prediction of secondary and three-dimensional structure of proteins. Moreover, she is currently applying virtual screening methods in drug repurposing studies. Dr Xxxxxxxxxx has published 155 long publications, 25 abstract/minor publications, and a chapter in Metabolomics book. She has been guest editor in three special issues on IJMS journal.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx	Experimental Pharmacology Unit, Mercogliano Laboratory	Mercogliano (Avellino), Italy	Staff Investigator	2022	2022
IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx	Experimental Pharmacology Unit, Mercogliano Laboratory	Mercogliano (Avellino), Italy	Researcher	2020	2022
IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx	Cancer Research Center of Mercogliano (CROM)	Mercogliano (Avellino), Italy	Senior Fellow	2008	2019
National Research Council (CNR)	Institute of Food Science - Laboratory of Computational Biology and Bioinformatics	Avellino, Italy	Fellow	2005	2008
Second University of Naples	Interdepartmental Research Center of Computational Sciences (C.R.I.S.C.E.B.)	Naples, Italy	PhD student	2000	2004

Other awards and honors

- 2005 to 2016: Faculty Member & PhD in Computational Biology at Second University of Naples

- 2014 - 2023: Scientific enable for associate professor (05 / E2 - Molecular Biology and 05 / E1 - General Biochemistry and Clinical Biochemistry), and full professor (05 / E1 - General Biochemistry and Clinical Biochemistry)

- Dr. Xxxxxxxxxx is currently Professor of Biochemistry and Clinical Biochemistry and Molecular Biology at Nursing Course of University of Xxxxxx Xxxxxxxx XX

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
Ministero della Salute - RIcerca Corrente	IRCCS Istituto Nazionale Tumori Fondazione X Xxxxxxx	2022	Definizione di nuove strategie terapeutiche di combinazione per potenziare l'efficacia del trattamento con anti-EGFR e prevenirne/ritardarne i meccanismi di resistenza nei tumori colorettali (Linea 3/2)	Collaborator	30.000,00	NA
Ministero della Salute - Ricerca Corrente	IRCCS Istituto Nazionale Tumori Fondazione X Xxxxxxx	2022	Caratterizzazione dell'impatto del microbioma in modelli murini di carcinoma gastrico durante i trattamenti chemioterapici ed immunoterapici (Linea 3/12)	Collaborator	15.000,00	NA
Ministero della Salute - Ricerca Corrente	IRCCS Istituto nazionale Tumori Fondazione X Xxxxxxx	2022	Targeting molecolare dei recettori beta adrenergici come strategia per il superamento della resistenza nei tumori testa collo( HNSCC) (Linea 4/1)	Collaborator	15.000,00	NA

Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
Ministero della Salute - Ricerca Corrente	IRCCS Istituto Nazionale Tumori Fondazione X Xxxxxxx	2022	I Macrofagi associati al tumore (TAM) e le cellule soppressorie di derivazione mieloide (MDSC) come bersagli terapeutici per riattivare la sorveglianza immunitaria nel carcinoma mammario triplo negativo (TNBC) (Linea 2/9)	Collaborator	25.000,00	NA
Ministero della Salute - Ricerca corrent	IRCCS Istituto Nazionale Tumori Fondazione X Xxxxxxx	2022	Ricerca Traslazionale ed applicazione di metodiche di Intelligenza Artificiale per la definizione di percorsi analgesici patient-centered nel trattamento del dolore oncologico da neoplasia del pancreas (Linea 4/4)	Collaborator	20.000,00	NA
Ministero della Salute - Ricerca Corrente	IRCCS Istituto Nazionale Tumori Fondazione X Xxxxxxx	2022	Ri-educare il Microambiente Tumorale disfunzionale per un approccio terapeutico innovativo del sarcoma (Linea 2/5)	Collaborator	30.000,00	NA
Ministero della Salute - Ricerca Corrente	IRCCS Istituto Nazionale Tumori Fondazione X Xxxxxxx	2022	Riposizionamento dell'antiepilettico acido valproico nel trattamento dei tumori metastatici colonrettali per potenziare l'efficacia della terapia standard e come immunomodulatore per prevenire/bypassare i meccanismi di resistenza al trattamento con inibitori di checkpoint immunitari (Linea 2/2)	Collaborator	35.000,00	NA
Ministero della Salute - Ricerca Corrente	IRCCS Istituto Nazionale Tumori Fondazione X Xxxxxxx	2022	Identificare fattori prognostici e predittivi di risposta al trattamento medico sistemico di I linea nei pazienti affetti da HCC avanzato (Linea 3/11)	Collaborator	15.000,00	NA
Ministero della Salute - Ricerca Corrente	Istituto Nazionale Tumori IRCCS Fondazione	2022	Evaluating the role of the selenoproteins and VCP/p97 as new prognostic markers and therapeuthic targets in solid cancers- Linea 1/4	Coordinator	20.000,00	xxxxx://xxxxxxxxxxxx.x xxx.xx/xxxxxxxxxxxx/x reaprogetti

2.7 Research Collaborators n. 6 - Under 40

Last Name: FATTORE

First Name: XXXXX

Last name at birth:

Gender: M

Title: Contribution to studies of in vitro cell lines and in vivo mouse models of drug resistance

Nationality: Italiana

Date of birth: 12/06/1985

Official H index (Scopus or Web of Science): 13.0

Country of residence: ITALY Country of Birth: ITALY Place of Birth: Maddaloni

Scopus Author Id:55200759600 ORCID ID:0000-0002-9245-0126 RESEARCH ID:K-7791-2016

Contact address

Current organisation name: Istituti fisioterapici ospitalieri - Istituto Regina Elena

Current Department / Faculty / Institute / Laboratory name: Scientific Direction - Preclinical Models and New Therapeutic

Agents

Street: Xxx Xxxx Xxxxxxxx 00

Postcode / Cedex: 00144 Town: Roma

Phone:x000000000000 Phone 2:

Education / training
Educational institution and location	Degree	Field of study	From year	To year
University "Sapienza", Rome, Italy	PhD	PhD in experimental Medicine - Study of the mechanisms of resistance to therapy in human cancers.	2011	2015
University of Naples "Federico II", Faculty of Biotechnology, Naples, Italy	Master's Degree / Laurea Magistrale	Study the role of WT1/ZNF224 interaction in the modulation of the expression of apoptotic genes in leukemia cells.	2007	2011
University "Xxxxx Xxxxxxxxxx", Faculty of Sciences MM. FF. NN, Caserta, Italy	Bachelor Degree / Laurea Triennale	Pathogenesis of prion diseases	2003	2007

Personal Statement:

Xxxxx Xxxxxxx has a long experience as collaborator in several projects focused on melanoma research. He is familiar with the statistical analyses of experimental data and in the extraction and evaluation of expression data from public databases (i.e. TCGA). He has a large experience in in vitro cell culture management, molecular biology techniques and drug efficacy studies. In last few years, his research activities have been focused on the study of microRNAs in melanoma aiming to identify novel therapeutics and predictors of drug resistance. He is familiar with in vivo experimental models and bioinformatics analyses. Dr. Fattore is co-inventor of an approved italian and PCT extended patent entitled "miRNAs for the in vitro diagnosis of drug resistant tumors".

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
IRCCS Regina Elena National Cancer Institute	SAFU Laboratory	Rome, Italy	Fixed Term Researcher	2020	2022
Istituto Nazionale Tumori IRCCS Fondazione Xxxxxxx	Department of Melanoma, Oncologic Immunotherapy and Innovative Therapies	Naples, Italy	Fixed term Researcher	2019	2020
Istituto Pasteur Italia- Fondazione Xxxxx Xxxxxxxxxx	Department of Molecular and Clinical Medicine, University "Sapienza"	Rome, Italy	Post doc	2019	2019
IRCCS Regina Elena National Cancer Institute	Preclinical Models and New Therapeutic Agents Unit	Rome, Italy	Post doc with Fondazione Xxxxxxx Xxxxxxxx fellowship	2018	2018
Ohio State University	Department of Molecular Virology Immunology and Medical Genetics	Columbus, Ohio	Research collaborator	2016	2016

Other awards and honors

Recipient of the research projects: "Xxxxxx Xxxxxxx"

(Fondazione Cenci Bolognetti). Fellowships: Fondazione Veronesi, AIRC "Xxxxxxxx Xxxxxxx" and "Xxxxxxxx Xxxxx"; Oral Communications at meetings: SiBBM 2016 and 2017; SIC 2013; Melanoma Bridge 2013, 2020 and 2021; best poster prize at IMI virtual congress 2020.

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
LILT	IRCCS Regina Xxxxx National Cancer Institute	2021	Therapeutic and diagnostic implications of miR-4488 and miR-4443 to fight resistance to targeted therapy in metastatic melanoma	Coordinator	50.000,00	xxxxx://xxx.xxxx.xx/xxxxx /default/files/inline- files/Elenco%20per% 20sito.pdf

2.8 Research Collaborators n. 7 - Under 40

Last Name: DI MARTILE

First Name: XXXXX

Last name at birth:

Gender: F

Title: In vitro and in vivo preclinical studies with LNP.miRs formulations

Nationality: ITALIANA

Date of birth: 02/11/1988

Official H index (Scopus or Web of Science): 11.0

Country of residence: ITALY Country of Birth: ITALY Place of Birth: Chieti

Scopus Author Id:55915357300 ORCID ID:0000-0001-9924-9022 RESEARCH ID:K-3348-2018

Contact address

Current organisation name: Istituti fisioterapici ospitalieri - Istituto Regina Elena

Current Department / Faculty / Institute / Laboratory name: Scientific Direction - Preclinical Models and New Therapeutic

Agents

Street: Xxx Xxxx Xxxxxxxx 00

Postcode / Cedex: 00144 Town: Roma

Phone:x000000000000 Phone 2:

Education / training
Educational institution and location	Degree	Field of study	From year	To year
Sapienza University, Azienda Ospedaliera Sant'Xxxxxx, Rome, Italy	Specialization / Specializzazione	Immunology, Pathology and Laboratory Medicine	2019	2022
Sapienza University, Rome, Italy	PhD	Histone AcetylTransferase (HAT) inhibitors as therapeutic approach affecting lung cancer growth and angiogenesis	2014	2018
Tor Vergata University, Rome, Italy	Master's Degree / Laurea Magistrale	The role of BH4 domain and cellular localization of bcl-2 protein in tumor progression and angiogenesis	2011	2013
Tor Vergata University, Rome, Italy	Bachelor Degree / Laurea Triennale	Induction of apoptosis by ionizing radiation in cultured human epithelial cells (HaCat)	2007	2011

Personal Statement:

Dr. Di Martile has experience as a PI of a young researcher xxxxx funded by IRE and as a collaborator of several funded research projects (AIRC IG, LILT). She has a large experience in in vivo studies (FELASA certification) and she has been in charge of performing experiments in approved Ministry of Health studies (n° 342/2019-PR 06/05/2019; n° 563/2021 27/07/2021). She has also a large experience in in vitro cell culture management, molecular biology techniques and drug efficacy studies. In last years, her research activities have been focused on the dissection of the crosstalk between tumor, stromal and immune cells. She has been also experienced in the evaluation of efficacy of anticancer agents (such as epigenetic drugs) in preclinical models of different tumor histotypes.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
IRCCS Regina Elena National Cancer Institute	Preclinical Models and New Therapeutic Agents Unit	Rome, Italy	Fixed-term researcher	2020	2022
IRCCS Regina Elena National Cancer Institute	Preclinical Models and New Therapeutic Agents Unit	Rome, Italy	Post-doc triennal AIRC/FIRC fellow	2020	2022
IRCCS Regina Elena National Cancer Institute	Preclinical Models and New Therapeutic Agents Unit	Rome, Italy	Post-doc fellow	2018	2019
IRCCS Regina Elena National Cancer Institute	Preclinical Models and New Therapeutic Agents Unit	Rome, Italy	PhD student fellow	2014	2017
IRCCS Regina Elena National Cancer Institute	Experimental Chemotherapy Laboratory	Rome, Italy	Research collaborator	2013	2014

Other awards and honors

2020-2022: Triennal AIRC/FIRC Fellowship ""Xxxxxxx Xxxxxxxxxxx Xxxxxxxx Xxxxxxx""

2018-2020: PI of three year Young researcher grant by IRCCS Xxxxxx Xxxxx National Cancer Institute 2018: Travel bursary to attend SIC Meeting, Milan.

2018: Travel bursary to attend XXXX Xxxxxxx, Xxxxxxx. 0000: Travel Bursary to attend EACR Meeting, Manchester 2013: Best poster Prize at the SIC Meeting, Catanzaro.

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
LILT	IRCCS Regina Xxxxx National Cancer Institute	2021	Therapeutic and diagnostic implications of miR-4488 and miR-4443 to fight resistance to targeted therapy in metastatic melanoma	Collaborator	50.000,00	xxxxx://xxx.xxxx.xx/xxxxx /default/files/inline- files/Elenco%20per% 20sito.pdf
Ministero della Salute - Ricerca Corrente	IRCCS Regina Elena National Cancer Institute	2018	L'inibizione di bcl-2 come nuova strategia terapeutica per contrastare la progressione del melanoma ed incrementare la risposta al trattamento farmacologico	Coordinator	65.000,00	xxxxx://xxxxxxxxxxxx.x xxx.xx/xxxxxxxxxxxx/x reaprogetti

2.9 Additional Research Collaborators n. 2 - Under 40 to hire

Last Name: Xxxx

First Name: Xxxxxxx

Last name at birth: Xxxx

Gender: F

Title: Development of lipid nanoparticles encapsulating miRNAs

Nationality: Italiana

Date of birth: 03/04/1992

Official H index (Scopus or Web of Science): 6.0

Country of residence: ITALY Country of Birth: ITALY Place of Birth: Napoli

Scopus Author Id:57192379338 ORCID ID:0000-0002-7263-7209 RESEARCH ID:AHD-7836-2022

Contact address

Current organisation name: Università degli Studi di Xxxxxx Xxxxxxxx XX

Current Department / Faculty / Institute / Laboratory name: Department of Pharamcy

Street: Dipartimento di Farmacia - Università degli Studi di Xxxxxx Xxxxxxxx XX, via X. Xxxxxxxxx, 49

Postcode / Cedex: 80131 Town: Napoli

Phone:x000000000000 Phone 2:

Education / training
Educational institution and location	Degree	Field of study	From year	To year
Imperial College London - Department of Materials - South Kensington Campus Xxxxxx XX0 0XX, XX	PhD	Design and development of smart materials for biomedical applications	2015	2020
University of Xxxxxx Xxxxxxxx XX - Scuola Politecnica e delle Scienze di Base- Xxx Xxxxxxx, 00, 00000 Xxxxxx, XX	Master's Degree / Laurea Magistrale	Chemical Engineering	2013	2015
University of Xxxxxx Xxxxxxxx XX - Scuola Politecnica e delle Scienze di Base- Xxx Xxxxxxx, 00, 00000 Xxxxxx, XX	Bachelor Degree / Laurea Triennale	Chemical Engineering	2010	2013

Personal Statement:

Currently a Fondazione Veronesi Post-doctoral Fellow working on the development of self-assembling nanoparticles encapsulating microRNA for the treatment of metastatic melanoma. Has an established track-record comprising 15 peer- reviewed publications, 1 filed patent, 6 presentations at international conferences, and numerous early career awards. She has investigated liposome-based formulations for the design of ultrasound-responsive systems and has developed a platform that enables the use of ultrasound as a novel trigger for enzymatic catalysis and hydrogel formation. Dr Xxxx has also mastered the use of neutron and X-ray scattering techniques, as well as cryo-TEM.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
University of Xxxxxx Xxxxxxxx XX	Drug Delivery Group - BioNanoMed	Xxx Xxxxxxxx Xxxxxxxxx 00, Xxxxxx (Xxxxx)	Fondazione Veronesi Post- Doctoral Fellow	2022	2022
University of Xxxxxx Xxxxxxxx XX	Drug Delivery Group - BioNanoMed	Xxx Xxxxxxxx Xxxxxxxxx 00, Xxxxxx (Xxxxx)	Post-Doctoral Researcher	2020	2020
Imperial College London	Xxxxxxx Group - Department of Materials	South Kensington Campus Xxxxxx XX0 0XX, XX	PhD Student	2015	2020

Other awards and honors

2022 Adritelf Travel Award € 500

2020 Italy Made Me Award conferito dall¿Ambasciata d¿Italia a Londra a ricercatori italiani che operano nel Regno Unito in riconoscimento di ricerca innovativa condotta in ciascuno dei tre settori ERC £ 1000.

2020 Matthey PhD Prize dal Department of Materials, Imperial College London £ 250. Travel grants for a total of £ 1300

Primo premio come migliore laureata del corso di Laurea Magistrale in Ingegneria Chimica. A/a 2015-2016, Università Xxxxxxxx XX € 1.000

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
Fondazione Veronesi	University of Xxxxxx Xxxxxxxx XX	2022	Sanp as novel nucleic acids vectors for precision medicine against melanoma	Coordinator	30.000,00	NA
Imperial College London	Imperial College London	2019	Beamtime application at the ISIS Neutron and Muon Source (UK) - RB1920464 "Effect of formulation method and lipid composition on liposome lamellarity	Collaborator	1.000,00	NA
Imperial College London	Imperial College London	2019	Beamtime application at the ISIS Neutron and Muon Source (UK) - RB1910285 ¿Self-assembly of thermo- responsive gels	Collaborator	1.000,00	NA
Imperial College London	Imperial College London	2018	Beamtime application at the ISIS Neutron and Muon Source (UK) - RB1810203 "Self-assembly of lipid and triblock copolymer-based amphiphyles for drug delivery"	Collaborator	1.000,00	NA
Imperial College London	Imperial College London	2018 - 2019	Rosetrees Trust PhD Plus Award	Coordinator	18.700,00	NA
Imperial College London	Imperial College London	2015 - 2018	Xxxxxxxxxxx Xxxxx Founder's Scholarship	Coordinator	96.000,00	NA

2.10 Additional Research Collaborators n. 3 - Under 40 to hire

Last Name: Affatato

First Name: Xxxxxxx

Last name at birth:

Gender: F

Title: Development of PDX models

Nationality: ITALIANA

Date of birth: 06/05/1984

Official H index (Scopus or Web of Science): 7.0

Country of residence: ITALY Country of Birth: ITALY Place of Birth: FOGGIA

Scopus Author Id:56728783300 ORCID ID:0000-0002-2235-039X RESEARCH ID:AHD-9757-2022

Contact address

Current organisation name: IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx

Current Department / Faculty / Institute / Laboratory name: Dipartimento di Ricerca Traslazionale a supporto dei percorsi

oncologici/Experimental Pharmacology Unit/ Mercogliano Systems Biology and Bioinformatics Laboratory

Street: Xxx Xxxxxxx Xxxxxxx 00

Postcode / Cedex: 80131 Town: NAPOLI

Phone:x000000000000 Phone 2:

Education / training
Educational institution and location	Degree	Field of study	From year	To year
Istituto di Ricerche Farmacologiche Xxxxx Xxxxx-IRCCS	PhD	PhD in Pharmacological Sciences: New therapeutic targets in ovarian cancer; mechanisms of resistance to therapy and PDX models	2017	2019
University of Parma	Master's Degree / Laurea Magistrale	Antitumor effects of retinoids in breast cancer	2008	2012

Personal Statement:

Dr Affatato research activity is currently focused on the study of mechanisms of resistance to anti-EGFR therapy in KRAS wt/BRAF wt and KRAS wt/BRAF mut colorectal cancer. She has a large experience in in vitro cell culture, molecular biology and drug efficacy studies. In last years her research acitivties have been focused on establishment and characterization of Patient-derived Xenograft (PDX) models of different tumor types (ovary, mesothelioma, thymoma) and evaluation of efficacy of anticancer agents.

Positions and honors

Positions
Institution	Division / Research group	Location	Position	From year	To year
Istituto Nazionale Tumori IRCCS Fondazione X. Xxxxxxx	Experimental Pharmacology Unit	Naples, Italy	Post-doc Fellow	2020	2022
Istituto di Ricerche Farmacologiche Xxxxx Xxxxx- IRCCS	Oncology Department/Laboratory of Molecular Pharmacology	Milan, Italy	PhD student	2017	2019
Istituto di Ricerche Farmacologiche Xxxxx Xxxxx- IRCCS	Oncology Department/Laboratory of Molecular Pharmacology	Milan, Italy	Fellow	2016	2016
Istituto di Ricerche Farmacologiche Xxxxx Xxxxx- IRCCS	Laboratory of Cardiovascular Clinical Pharmacology	Milan, Italy	Fellow	2013	2015

Other awards and honors

N/A

Grant
Funded by Institution	Researcher inst. where xxxxx is/was performed	Year	Title	Position in Projects	Fund (euro)	Source website grant listed
N/A	N/A	N/A	N/A	Collaborator	0,00	N/A

2.17 Expertise Research Collaborators

Selected peer-reviewed publications of the Research Group / Collaborators
Collaborato	Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**	P.*
Nele Valeria	Investigation of the Thermogelation of a Promising Biocompatible ABC Triblock Terpolymer and Its Comparison with Pluronic F127	Article	1783- 1799	55	2022	10.1021/acs.macromol.1 c02123	35431333	1	O
Nele Valeria	Design of Lipid-Based Nanocarriers via Cation Modulation of Ethanol- Interdigitated Lipid Membranes	Article	11909- 11921	37	2021	10.1021/acs.xxxxxxxx.1c 02076	34581180	2	F
Xxxxxxx Xxxx	Circulating Vitamin D levels status and clinical prognostic indices in COVID-19 patients	Article	NOT_FO UND	22	2021	10.1186/s12931-021- 01666-3	33658032	22	O
Nele Valeria	Ultrasound-Triggered Enzymatic Gelation	Article	NOT_FO UND	32	2020	10.1002/adma.20190591 4	31922627	15	F
Xxxxxxxx Xxxxxxx	Identification of PLK1 as a new therapeutic target in mucinous ovarian carcinoma	Article	NOT_FO UND	12	2020	10.3390/cancers120306 72	32183025	14	F
del bufalo donatella	Inhibition of Anti-Apoptotic Bcl- 2 Proteins in Preclinical and Clinical Studies: Current Overview in Cancer	Review	NOT_FO UND	9	2020	10.3390/cells9051287	32455818	31	L
Nele Valeria	Effect of Formulation Method, Lipid Composition, and PEGylation on Vesicle Lamellarity: A Small-Angle Neutron Scattering Study	Article	6064- 6074	35	2019	10.1021/acs.xxxxxxxx.8b 04256	30977658	38	F
Xxxxxxxx Xxxxxxx	Overcoming platinum-acquired resistance in ovarian cancer patient-derived xenografts	Article	NOT_FO UND	11	2019	10.1177/1758835919839 543	31258626	19	O
Xxxxxxx Xxxx	Metabolic features of cancer stem cells: The emerging role of lipid metabolism	Article	2367- 2378	37	2018	10.1038/s41388-018- 0141-3	29445137	64	F
Xxxxxxx Xxxx	Inhibition of Stearoyl-CoA desaturase 1 reverts BRAF and MEK inhibition-induced selection of cancer stem cells in BRAF-mutated melanoma	Article	NOT_FO UND	37	2018	10.1186/s13046-018- 0989-7	30558661	33	L
Xxxxxxxx Xxxxxxx	Recent insights into mucinous ovarian carcinoma	Review	NOT_FO UND	19	2018	10.3390/ijms19061569	29795040	29	O
del bufalo donatella	Pharmacological activation of SIRT6 triggers lethal autophagy in human cancer cells	Article	NOT_FO UND	9	2018	10.1038/s41419-018- 1065-0	30250025	48	O

Collaborato	Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**	P.*
Xxxxxxxxxx Xxxxx	Xxxxxxxxx B1 and M1: Biological properties and their involvement in cancer development	Review	NOT_FO UND	10	2018	10.3390/toxins10060214	29794965	144	O
DI XXXXXXX XXXXX	Emerging role of histone acetyltransferase in stem cells and cancer	Review	NOT_FO UND	2018	2018	10.1155/2018/8908751	30651738	30	O
XXXXXXX XXXXX	Inhibition of Stearoyl-CoA desaturase 1 reverts BRAF and MEK inhibition-induced selection of cancer stem cells in BRAF-mutated melanoma	Article	NOT_FO UND	37	2018	10.1186/s13046-018- 0989-7	30558661	33	O
Xxxxxxxx Xxxxx Xxxxxxx	Overall Survival with Combined Nivolumab and Ipilimumab in Advanced Melanoma	Article	1345- 1356	377	2017	10.1056/NEJMoa170968 4	28889792	1921	O
Xxxxxxx Xxxx	Xxxxxxxx-CoA-desaturase 1 regulates lung cancer stemness via stabilization and nuclear localization of YAP/TAZ	Article	4573- 4584	36	2017	10.1038/onc.2017.75	28368399	79	L
Xxxxxxx Xxxx	Xxxxxxxx of Stearoyl-CoA- desaturase 1 activity reverts resistance to cisplatin in lung cancer stem cells	Article	93-104	406	2017	10.1016/j.canlet.2017.07 .027	28797843	61	L
DI XXXXXXX XXXXX	BCL-X<inf>L</inf> overexpression promotes tumor progression-associated properties article	Article	NOT_FO UND	8	2017	10.1038/s41419-017- 0055-y	29238043	46	O
XXXXXXX XXXXX	MicroRNAs in melanoma development and resistance to target therapy	Review	22262- 22278	8	2017	10.18632/oncotarget.147 63	28118616	00	X
XX XXXXXXX XXXXX	Histone acetyltransferase inhibitor CPTH6 preferentially targets lung cancer stem-like cells	Article	11332- 11348	7	2016	10.18632/oncotarget.723 8	26870991	00	X
XX XXXXXXX XXXXX	The multifaceted role of lysine acetylation in cancer: Prognostic biomarker and therapeutic target	Article	55789- 55810	7	2016	10.18632/oncotarget.100 48	27322556	47	F
XXXXXXX XXXXX	xxX-579-3p controls melanoma progression and resistance to target therapy	Article	E5005- E5013	113	2016	10.1073/pnas.16077531 13	27503895	71	F
Xxxxxxxx Xxxxx Xxxxxxx	Xxxxxxxxx versus chemotherapy in patients with advanced melanoma who progressed after anti-CTLA-4 treatment (CheckMate 037): A randomised, controlled, open- label, phase 3 trial	Article	375-384	16	2015	10.1016/S1470- 2045(15)70076-8	25795410	1851	O

Collaborato	Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**	P.*
Xxxxxxxx Xxxxx Xxxxxxx	Combined nivolumab and ipilimumab or monotherapy in untreated Melanoma	Article	23-34	373	2015	10.1056/NEJMoa150403 0	26027431	4638	O
De Xxxx Xxxxxxxx	Multifunctional polymeric micelles co-loaded with anti- survivin siRNA and paclitaxel overcome drug resistance in an animal model of ovarian cancer	Article	1075- 1084	14	2015	10.1158/1535- 7163.MCT-14-0556	25657335	76	O
Xxxxxxxx Xxxxxxx	All-trans-retinoic Acid Modulates the plasticity and inhibits the motility of breast cancer cells role of notch1 and transforming growth factor (TGF ß)	Article	17690- 17709	290	2015	10.1074/jbc.M115.63851 0	26018078	34	O
Xxxxxxxxxx Xxxxx	GPX4 and GPX7 over- expression in human hepatocellular carcinoma tissues	Article	5-10	59	2015	10.4081/ejh.2015.2540	26708178	46	L
Xxxxxxxx Xxxxx Xxxxxxx	Xxxxxxxx vemurafenib and cobimetinib in BRAF-mutated melanoma	Article	1867- 1876	371	2014	10.1056/NEJMoa140886 8	25265494	1380	O
Xxxxxxxx Xxxxx Xxxxxxx	Xxxxxxxxx in previously untreated melanoma without BRAF mutation	Article	320-330	372	2015	10.1056/NEJMoa141208 2	25399552	3735	L
De Xxxx Xxxxxxxx	Xxx-34: A new weapon against cancer?	Review	e195	3	2014	10.1038/mtna.2014.47	25247240	360	O
De Xxxx Xxxxxxxx	Transferrin-conjugated SNALPs encapsulating 2?-O- methylated miR-34a for the treatment of multiple myeloma	Article	NOT_FO UND	2014	2014	10.1155/2014/217365	24683542	49	L
De Xxxx Xxxxxxxx	Xxxxxxxxx micelles containing reversibly phospholipid- modified anti-survivin siRNA: A promising strategy to overcome drug resistance in cancer	Article	224-231	343	2014	10.1016/j.canlet.2013.09 .037	24099916	76	O
del bufalo donatella	1,3,4-Oxadiazole-containing histone deacetylase inhibitors: Anticancer activities in cancer cells	Article	6259- 6265	57	2014	10.1021/jm500303u	24972008	78	O
del bufalo donatella	VEGF-induced neoangiogenesis is mediated by NAADP and two-pore channel-2 -dependent ca<sup>2+</sup>signaling	Article	4706- 4715	111	2014	10.1073/pnas.14060291 11	25331892	107	O
DI XXXXXXX XXXXX	Histone deacetylase inhibition synergistically enhances pemetrexed cytotoxicity through induction of apoptosis and autophagy in non-small cell lung cancer	Article	NOT_FO UND	13	2014	10.1186/1476-4598-13- 230	25301686	44	O

Collaborato	Title	Type	Pag	Vol	Year	DOI	PMID	Cit.**	P.*
De Xxxx Xxxxxxxx	Xxxxxxxxxxxx for topical administration of resveratrol: A comparative study	Article	179-187	440	2013	10.1016/j.ijpharm.2012.0 8.009	22909994	110	L
Xxxxxxxxxx Xxxxx	Acquired resistance to zoledronic acid and the parallel acquisition of an aggressive phenotype are mediated by p38-MAP kinase activation in prostate cancer cells	Article	NOT_FO UND	4	2013	10.1038/cddis.2013.165	23703386	52	O
Xxxxxxxxxx Xxxxx	Gene expression signature of human HepG2 cell line	Article	335-345	518	2013	10.1016/j.gene.2012.12. 106	23357223	66	F
Xxxxxxxxxx Xxxxx	Functional and structural features of adipokine family	Review	1-14	61	2013	10.1016/j.cyto.2012.08.0 36	23022179	82	L
XXXXXXX XXXXX	Combination therapy with anti- ErbB3 monoclonal antibodies and EGFR TKIs potently inhibits non-small cell lung cancer	Article	1253- 1265	4	2013	10.18632/oncotarget.114 1	23896512	34	O
XXXXXXX XXXXX	Activation of an early feedback survival loop involving phospho-ErbB3 is a general response of melanoma cells to RAF/MEK inhibition and is abrogated by anti-ErbB3 antibodies	Article	NOT_FO UND	11	2013	10.1186/1479-5876-11- 180	23890105	52	F
del bufalo donatella	The mitogen-activated protein kinase (MAPK) cascade controls phosphatase and tensin homolog (PTEN) expression through multiple mechanisms	Article	667-679	90	2012	10.1007/s00109-011- 0844-1	22215152	54	O

* Position: F=First L=Last C=Correspondent O=Other N=Not applicable

** Autocertificated

3 - Ethics

1. HUMAN EMBRYOS/FOETUSES
Does your research involve Human Embryonic Stem Cells (hESCs)?	No
Does your research involve the use of human embryos?	No
Does your research involve the use of human foetal tissues / cells?	No
2. HUMANS
Does your research involve human participants?	Yes
Does your research involve physical interventions on the study participants?	Yes

3. HUMAN CELLS / TISSUES
Does your research involve human cells or tissues (other than from Human Embryos/ Foetuses?	Yes
4. PERSONAL DATA
Does your research involve personal data collection and/or processing?	No
Does your research involve further processing of previously collected personal data (secondary use)?	No
5. ANIMALS
Does your research involve animals?	Yes
6. ENVIRONMENT & HEALTH and SAFETY
Does your research involve the use of elements that may cause harm to the environment, to animals or plants?	No
Does your research deal with endangered fauna and/or flora and/or protected areas?	No
Does your research involve the use of elements that may cause harm to humans, including research staff?	No
7. DUAL USE
Does your research involve dual-use items in the sense of Regulation 428/2009, or other items for which an	No
8. EXCLUSIVE FOCUS ON CIVIL APPLICATIONS
Could your research raise concerns regarding the exclusive focus on civil applications?	No
9. MISUSE
Does your research have the potential for misuse of research results?	No
10. OTHER ETHICS ISSUES
Are there any other ethics issues that should be taken into consideration? Please specify	No

X

I confirm that I have taken into account all ethics issues described above and that, if any ethics issues apply, I will complete the ethics self-assessment and attach the required documents.

4 - Call-specific questions

Eligibility
I acknowledge that I am aware of the eligibility requirements for applying as specified in the Call- PNRRXXXX_M6/C2, and certify that, to the best of my knowledge my application is in compliance with all these requirements. I understand that my proposal may be declared ineligible at any point during the evaluation or granting process if it is found not to be compliant with these eligibility criteria.	X
I confirm that the proposal that I am about to submit draws substantially don’t repeat on an existing or recently finished GRANT funded.	X

Data-Related Questions and Data Protection (Consent to any question below is entirely voluntary. A positive or negative answer will not affect the evaluation of your project proposal in any form and will not be communicated to the evaluators of your project.)
For communication purposes only, the MoH asks for your permission to publish,in whatever form and medium, your name, the proposal title, the proposal acronym, the panel, and host institution, should your proposal be retained for funding.	X
Some national and regional public research funding authorities run schemes to fund MoH applicants that score highly in the MoH's evaluation but which can not be funded by the MoH due to its limited budget. In case your proposal could not be selected for funding by the MoH do you consent to allow the MoH to disclose the results of your evaluation (score and ranking range) together with your name, non- confidential proposal title and abstract, proposal acronym, host institution and your contact details to such authorities?	X
The MoH is sometimes contacted for lists of MoH funded researchers by institutions that are awarding prizes to excellent researchers. Do you consent to allow the MoH to disclose your name, non-confidential proposal title and abstract, proposal acronym, host institution and your contact details to such institutions?	X
The Ministry of Health occasionally could contacts Principal Investigators of funded proposals for various purposes such as communication campaigns, pitching events, presentation of their project's evolution or outcomes to the public, invitations to represent the Ministry of Health in national and international forums, studies etc. Should your proposal be funded, do you consent to the Ministry of Health staff contacting you for such purposes?	X
For purposes related to monitoring, study and evaluating implementation of MoH actions, the MoH may need that submitted proposals and their respective evaluation data be processed by external parties. Any processing will be conducted in compliance with the requirements of Regulation 45/2001.

5 – Description Project

Summary description

BRAF-mutated metastatic melanoma relapsing after targeted or immunotherapy is an aggressive disease with high unmet clinical need. Only available therapeutic option is standard chemotherapy with dacarbazine with limited life expectancy.

Three members of this team (UO1, UO2, UO4) have recently identified and patented the use of a subset of microRNAs (miRNAs) in combination with targeted therapy to revert drug resistance (PCT/IT2019/050073: miRNAs for treatment and diagnosis of drug resistant tumors). The claims of this patent include systemic delivery of oncosuppressor miRNAs using lipid nanoparticles (LNPs). Our joint teams have generated in vitro and in vivo data demonstrating the antitumor efficacy of xxXXX-loaded LNPs (LNP-miRs) in combination with targeted therapy in melanoma models. This project aims at optimizing LNP-miRs composition and completing pre-clinical efficacy and preliminary toxicology in preparation to clinical translation into Phase I-II studies.

Background / State of the art

Targeted therapies with inhibitors of BRAF and MEK kinases (MAPKi) elicit prompt responses in most melanoma patients harboring BRAF mutations (Xxxx et al 2017). However, therapeutic efficacy is not durable, with a median duration of response of about one year due to acquired resistance (Xxxxxx et al 2019). Moreover, patients relapsing after MAPKi have a low response rate to salvage immunotherapies with Immune Checkpoint Inhibitors (ICIs) (Xxxx et al, 2021). In the last years, our group has focused on miRNAs as orchestrators of non-genetic mechanisms adopted by melanoma cells to challenge MAPKi therapy: we initially identified a panel of miRNAs able to halt the development of resistance to targeted therapies in BRAF-mutated melanoma (Fattore et al 2018). This discovery was patent protected (PCT/IT2019/050073).

Afterwards, confirmatory results were obtained by delivering specific oncosuppressors miRNAs (namely miR-204-5p, miR- 199b-5p, miR-579-3p) into melanoma cells relying on the capability of simultaneously targeting multiple oncogenic pathways. This was achieved through miRNAs encapsulation in LNPs to circumvent the main drawbacks of delivering naked RNAs, i.e. poor cellular uptake, off-target activity and nuclease degradation. Exploiting several drug sensitive and resistant in vitro melanoma models, we demonstrated that LNPs carrying miR-204-5p and miR-199b-5p were able to reduce cell proliferation alone and, more efficiently, when combined with MAPKi (Fattore et al 2020).

Description and distribution of activities of each operating unit

We believe that our multidisciplinary team has the required complementarity to successfully manage all the activities and reach the objectives planned in the project. "TACTIC" activities are organized into 3 aims (described in detail in the appropriate sections) and an outline of tasks for each collaborator with a clear indication of the role of each UO is reported below.

The first aim plans to establish the optimal formulation parameters for LNP-miRs and prepare to industrial scale-up. The UO3 will be in charge for this task. These activities will pave the way for the further clinical translation of LNPs-loaded oncosuppressive miRNAs for melanoma. Given that the use of LNP-miRs is necessary to achieve the goals of Aims 2 and the activities carried out by UO3 are transversal to the entire project.

The second aim will be carried out by UO1 and UO2. In particular, the UO1 will be in charge of the in vitro and in vivo preclinical studies with LNP-miR formulations in the attempt to a) characterize and b) determine the basal toxicity of LNP formulations as well as c) carry out the efficacy studies of LNPs in combination with targeted therapies. This will be accomplished on different drug resistant preclinical models already available in the labs and on which the team members has gathered large experience in the recent years. In parallel, UO2 will be in charge of the biochemical and histological analyses to detect potential alterations of mice organ functionality upon LNPs' treatments, together with the evaluation of tumor masses by immunohistochemistry (IHC), Western Blot and qRT-PCR analyses for expression levels of miRNAs and their target genes.

The third aim will be carried out by the UO4. The availability of samples derived from melanoma patients necessary for the development of PDX models will be guaranteed by UO4 daily clinical activities. Thanks to his renowned commitment to melanoma care and translational research, certified by many ongoing clinical trials, UO4 is a reference center for melanoma treatment, with thousands dermatological and oncological outpatient visits, thousands of Day Hospital cancer treatments, as well as, hundreds surgical patients every year. Most importantly, it is also equipped with a qualified animal facility and researchers with long lasting experience in in vivo studies. UO4 will also carry out genomic and transciptomic characterization of established PDX, in collaboration with the UO1.

In order to ensure efficient research collaboration, all the UO will meet quarterly to assess the work progress and make sure that the project tasks are completed in due course. All the data generated throughout the project will be made available to the team members on a shared data management platform and a common naming convention will be used to make the data findable and avoid potential errors. These measures will be instrumental for an effective project implementation.

The PI will supervise the entire project, monitoring the proper advancement, and controlling the connection between work plan and results obtained. In collaborations with co-PI and other principal collaborators, XX will write scientific papers.

5.4 Specific Aims and Experimental Design

Specific aim 1

LNPs encapsulating various combination of three oncosuppressor miRNAs, i.e. miR-204-5p, miR-199b-5p and miR-579-5p, will be prepared and fully characterized through well established protocols. Formulations will be based on an ionizable cationic lipid, neutral lipids (e.g. cholesterol and distearoylphosphatidylcholine), and a PEGylated lipid, combined to achieve the highest miRNAs encapsulation, stability in serum and high transfection efficiency on melanoma cells. LNP-miRs will be prepared by a well-established ethanol injection method followed by extrusion. Briefly, an ethanolic lipid solution containing

the ionizable lipid, cholesterol, DSPC and the PEGylated lipid at fixed molar ratios will be mixed with an aqueous miRNAs solution in citric acid. The obtained LNP suspension will be extruded through 200/100 nm pore-sized membranes with a thermobarrel extruder. The LNP-miRs formulation will be dialyzed against citrate buffer and against HEPES buffered saline (HBS) overnight to remove ethanol and to neutralize the LNP surface charge. The unencapsulated miRNAs will be then removed via ultracentrifugation. LNP-miRs formulations will be sterilized for the in vitro and in vivo studies. LNP-miRs hydrodynamic diameter, xxxxxxxxxxxxxx index, and zeta potential will be characterized by photon correlation spectroscopy (PCS) and electrophoretic light scattering, respectively. The nanoparticle stability against aggregation in serum will also be assessed via PCS at different time points while the miRNAs encapsulation efficiency will be quantified by spectrophotometry and calculated as the % ratio between the miRNAs actual loading (mg of miRNAs/mg of total lipids) and xxXXX theoretical loading in formulation. Finally, the LNP-miRs stability against aggregation in complex fluid and haemolysis assays on fresh human blood will be performed as previously reported (Campani et al, 2020).

Formulations characterized by homogeneous size below 200-250 nm, high miRNAs encapsulation efficiency, and stability in serum, will be then tested on the different melanoma preclinical models described in the following aims.

In order to facilitate the clinical translation and scale-up of LNP-miRs formulations, we will test microfluidic mixing as an alternative formulation method that ensures greater reproducibility compared to other ethanol injection methods.

Furthermore, microfluidic mixing allows a rapid scale-up of the achievable formulation volume by simple parallelization of the microfluidic mixers (Xxxxxxxx et al, 2019). We will use a microfluidic system equipped with a proprietary micromixer chip to produce LNP-miRs. The mixing parameters (i.e., the flow ratio between the ethanolic and the aqueous phases and the total flow rate) will be optimized to ensure a narrow size distribution (xxxxxxxxxxxxxx index < 0.2), final colloidal dimensions ¿ 200 nm, and high miRNAs entrapment efficiencies (> 85 %). We will also investigate the possibility of replacing dialysis and ultracentrifugation with tangential flow filtration (TFF) for buffer exchange and LNP purification. Both dialysis and ultracentrifugation are sub-optimal purification techniques for formulation volumes > 10 mL since they are time-consuming (dialysis) and can lead to potential sample loss and nanoparticle aggregation (ultracentrifugation). TFF may be a valid alternative for LNP purification since it enables large volumes processing with minimal sample loss (Xxx et al, 2021). We will set up a TFF purification protocol for LNP-miRs by using a TFF cassette equipped with a 50 kDa membrane (e.g., Minimate from Pall Corporation) and assess the extent of sample recovery by measuring the LNP concentration.

The work carried out to achieve Aim 1 will establish the optimal formulation parameters for LNP-miRs and will pave the way for further clinical translation and industrial scale-up of the formulation.

Specific aim 2

LNPs formulations for miRNAs delivery in melanoma cells will be tested in three different drug resistant preclinical models: namely in vitro, in vivo and ex vivo. As mentioned above, we will use combinations of three oncosuppressor miRNAs, i.e. miR-204-5p, miR-199b-5p and miR-579-5p in order to identify the most potent to be carried forward. As to in vitro models, we have already selected A375DR and LOX IMVIDR melanoma cell lines for their double resistance (DR) to both BRAF and MEK inhibitors. The collection of DR cells will be also expanded with patients' derived BRAF-mutated melanoma cell lines. First of all, we will test the capability of LNP-miRs to efficiently deliver their cargos to melanoma cells through qRT- PCR. The effect of miRNAs on known target genes both at RNA and protein levels will be evaluated by PCR and western blot analysis, respectively. Most importantly, through MTT and ATP detection assays we will determine the capability of LNP-miRs to affect cell viability of resistant melanoma cells and to revert drug resistance when combined with BRAFi and MEKi. Then, we will move to the testing in in vivo models. The first step will be to evaluate the potential toxicity, immunogenicity and side effects of LNPs in wild type mice (i.e. C57BL/6). These aspects will be determined as follows: 1) acute toxicity, i.e. possible adverse effects after first exposure to a single dose of LNPs; 2) maximum tolerated dose (MTD),

i.e. the highest single dose regimens showing no significant weight loss (<10%) and absence of adverse effects; 3)

repeated-dose toxicity, i.e. testing whether toxicity occurs after continuous exposure to LNPs. Furthermore, the serum levels of glutamate pyruvate transaminase (GPT), creatinine (CRE), and creatine kinase-MB (CK-MB) will be determined, together with biochemical and histological analyses to detect potential alterations of organ functionality (i.e. liver, kidney,

intestine and lung) (Xxxxxxxx et al. 2011; Xxxxxx et al. 2013; Lo et al. 2020). Afterwards, we will test the capability of therapeutic LNPs to revert drug resistance. To this aim, we will establish xenografts of resistant melanoma models by injecting A375DR or LOX IMVIDR in immunocompromised nude mice (i.e. CD1). When the tumors reach a volume of 150 mm3, mice will be treated with xxXXX-loaded LNPs alone or in combination with BRAFi and MEKi. Treatments will be performed for four weeks and tumor growth will be measured twice a week through non-invasive imaging. We will test different LNP-miRs dosing schedules (e.g. once, twice or three times weekly) in order to identify the most potent one. The primary outcome will be to measure objective responses (e.g. tumor regressions, stable response or progressive disease). After mice sacrifice, tumor masses will be collected and analysed by IHC, Western Blot and qRT-PCR analyses, to determine expression levels of the miRNAs and their target genes. Some in vivo studies will be extended to measure the ability of therapeutic combinations of targeted therapy and LNP-miRs to impair tumor regrowth. Finally, these results will be strengthened by establishing ex vivo cultures of drug resistant melanoma biopsies using fresh biopsies from melanoma patient (Xxxxxxxxx et al, 2016). Briefly, 300 micron meter thin slices will be exposed to the therapeutic LNPs alone or in combination with MAPKi and the proliferative potential of the cells inside the tumor material will be determined by measuring Ki67 and EdU incorporation by immunostaining. This model can also allow studying the immune infiltrate composing the tumor microenvironment (TME), which is of great interest given the capability of the three microRNAs to inhibit the expression of proinflammatory factors responsible for the maintenance of an immunosuppressive and drug resistant TME. Altogether, the results of this section will demonstrate the therapeutic capability of LNPs delivered miRNAs to revert MAPKi resistance in BRAF-mutated melanomas.

Specific aim 3

This aim will be directed to validate the therapeutic potential of LNP-miRs by exploiting PDX models. Melanoma PDX successfully recapitulate the tumor architecture and genotype of the original tumor by implanting fresh materials from patients directly into immunodeficient NOD-scid IL2Rgammanull (NSG) (Xxxxxx et al, 2016). The development of PDX models will be carried out at Istituto Xxxxxxx in compliance with institutional guidelines and regulations, after approval by the Italian Ministry of Health. Xxxxxxx is a well-known national reference center for melanoma care and translational research, also equipped with a qualified animal facility and researchers with long lasting experience in in vivo studies.

Ideally, it will be preferable to use samples coming from subcutaneous metastases that are more easily operable as compared to visceral metastasis. We plan to develop two different PDX models derived from BRAF mutated melanoma patients: 1) before starting BRAFi and MEKi therapy and 2) after progression from targeted therapies or immunotherapy (determined by RECIST criteria). In both models, tumor fragments will be implanted subcutaneously into the flanks of NSG mice for an initial propagation until tumor masses reach about 000-0000 mm3 in size. Then, tumors will be harvested and PDX will be 1) bio-banked and cryopreserved, 2) subjected to transcriptomic and genomic characterization (i.e. RNA-seq and Whole Exome Sequencing [WES], respectively) and 3) dissected again for the re-implantation and propagation in additional mice. These last mouse models will be subjected to drug efficacy studies. To this aim, mice will be treated with LNP-miRs alone or in combination with BRAFi and MEKi as reported in the previous task. We envisage two types of studies: pharmacokinetic-pharmacodynamic studies and efficacy studies. The first ones will be carried out in a small number of mice and will have a short duration. Their goal will be: a) to assess the amount and kinetic of miRNAs delivery to the tumour site at different time points after i.v. injection of LNP-miRs; b) to assess the degree of downregulation of selected miRNAs targets at the level of the PDX-derived tumour lesions. Efficacy studies will be carried out according to standardized protocols established with stable cells lines described in Aim2. The outcomes of the studies will be: 1) for PDX derived from drug naïve metastasis, to demonstrate the therapeutic capability of LNPs to potentiate targeted therapies and

2) for PDX derived from drug resistant biopsies, to evaluate the therapeutic capability of miRNAs to restore drug sensitivity.

After mice sacrifice, tumor masses will be collected and analyzed as described above to study not only the expression levels of miRNAs and their target genes, but also the activated cell death pathways and the TME status along with the composition of immune infiltrate. At the moment, we have available two PDX models for each condition (sensitive vs resistant biopsies), but we are confident that we will be able to successfully enlarge our collection given that the success rates for melanoma is very higher (about 63%) (Xxxxxx et al, 2017) compared to other solid tumor types.

In summary, we believe that the results obtained with these in vivo models, will conclude our pre-clinical proof-of concept anti-tumor efficacy and preliminary safety studies and will support transition to Phase I-II studies in BRAF-mutant melanoma patients.

Experimental design aim 1

Within aim 1, we will prepare LNPs encapsulating the three oncosuppressor miRNAs (i.e., miR-204-5p, miR-199b-5p, and miR-579-5p) in various ratios. The LNP formulation comprises a mixture of an ionizable cationic lipid (1,2-dioleoyl-3- dimethylammonium-propane, DODAP), cholesterol, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and N-palmitoyl- sphingosine-1-{succinyl[methoxy(polyethylene glycol)2000]} (Cer16PEG2000) at 20:45:25:10 molar ratio. miR-loaded LNPs (LNP-miRs) will be prepared by mixing an ethanolic solution containing the lipids in the appropriate ratios with an aqueous xxXXX solution in 20 mM citric acid. Prior to mixing, the lipid and the xxXXX solutions will be kept at 65 °C for 2 minutes.

The obtained LNP suspension will be extruded through 200 and 100 nm pore-sized polycarbonate membranes at 65 °C with a thermobarrel extruder. The final miR concentration in the formulation mix will be comprised between 200 and 400 µg/mL. The LNP-miR formulations will be first dialyzed against citrate buffer pH 4 (1 hour) and then against HEPES buffer pH 7.4 (overnight) to remove ethanol and to neutralize the LNP surface charge. The unencapsulated xxXXX will be removed via ultracentrifugation. Photon correlation spectroscopy (PCS) and electrophoretic light scattering will be used to determine the hydrodynamic diameter, the xxxxxxxxxxxxxx index (PDI), and the zeta potential of the formulations. The nanoparticle stability against aggregation in serum will also be assessed via PCS at different time points (e.g., 30 minutes and 4 hours) by diluting the LNP-miRs in serum to a final concentration of 1% wt/v. The xxXXX encapsulation efficiency will be quantified by spectrophotometry and calculated as the % ratio between the xxXXX actual loading (mg of xxXXX/mg of total lipids) and xxXXX theoretical loading in formulation. More specifically, an aliquot of LNP-miRs will be diluted in methanol at a 1:100 v/v ratio and centrifuged for 30 min at 13,000 rpm; the concentration of the miRNAs in the supernatant will be obtained by measuring the absorbance at 260 nm. The total lipid content of the formulations will be determined by the Xxxxxxx assay according to previously published protocol (Fattore, Campani et al 2020). Hemolysis assays on fresh human blood will be performed on LNP-miRs to assess their suitability for intravenous injection according to previously published protocols (Campani et al, 2020). Formulations characterized by hydrodynamic diameters up to 250 nm, up to PDI 0.25, xxXXX encapsulation efficiencies above 85%, good stability against aggregation in serum, and hemolysis below 5% will be tested on the different melanoma preclinical models described in the following aims.

We will also test microfluidic mixing as an alternative formulation method; more specifically, we will use a Dolomite®

microfluidic system equipped with a proprietary micromixer chip which allows the rapid mixing of the ethanolic phase containing the lipids and the aqueous phase containing the miRs. We will test three different flow ratios between the two phases (1:1.5, 1:3, 1:5) and total flow rates of 120 and 160 µL/min. Also in this case, we will use PCS and electrophoretic light scattering to assess the hydrodynamic diameter, the PDI, and the zeta potential of the obtained formulations and spectrophotometry to determine the miRs encapsulation efficiencies. Targets include a narrow size distribution (xxxxxxxxxxxxxx index < 0.25), final colloidal dimensions below 250 nm, and high xxXXX xxxxxxxxxx efficiencies (above 85

%). We will also evaluate the using tangential flow filtration (TFF) instead of dialysis and ultracentrifugation for buffer exchange and LNP purification. We will first use plain LNPs to identify the optimal flow rate and feed pressure values which ensure a sample recovery above 75% and minimal variations in the colloidal dimensions of the formulations. The use of TFF for LNP purification will be then tested on LNP-miR formulations.

Experimental design aim 2

This aim will be structured to test the therapeutic efficacy of LNP-miRs, developed in the previous aim, in different drug resistant melanoma models. Starting from the in vitro setting, we will expand the collection of melanoma cells double resistant (DR) to both BRAFi and MEKi, in addition to the already available A375DR and LOX IMVI DR cell lines. To establish DR cells, at 2-3 patients' derived BRAF-mutated melanoma cell cultures, will be exposed to increasing

concentrations of BRAFi starting from 2 and 1nM, respectively. The selection will last until the concentrations of 2 uM for the BRAFi and 1 uM for the MEKi is reached. The acquisition of drug resistance will be assessed in cell proliferation assays using the sensitive counterparts as controls. These cell lines will be characterized for 1) the down-regulation of the oncosuppressive miRNAs (qRT-PCR), and 2) upregulation of their known target genes (e.g. BCL-2 for miR-204-5p, VEGFa for miR-199b-5p and BRAF for miR-579-5p); 3) RNA sequencing. We will then test if LNP-miRs are able to efficiently deliver their cargos to melanoma cells and to inhibit the expression of the target genes as previously described for other drug resistant cell lines. Their effects con cell viability will be then evaluated. To this purpose, MAPKi-resistant cells will be exposed to LNP-miRs for 72h to perform in vitro clonogenic, MTT and ATP detection assays. Furthermore, we will test if LNP-miRs are able to potentiate the efficacy of MAPKi testing their ability to delay the onset of resistance using long term

(4-5 weeks) in vitro clonogenic assays.

We plan to validate these data by exploiting ex vivo melanoma metastasis cultures as described by Xxxxxxxxx et al, 2016. To this purpose, fresh biopsies from patients relapsing after MAPKi will be sectioned, placed in culture media and then incubated with LNP-miRs + MAPKi at increasing concentrations for 5 days. The efficacy of different treatments will be evaluated immunostaining tumor sections for Ki67 and EdU incorporation.

Regarding the in vivo setting, a first step will be to evaluate the potential toxicity, immunogenicity and side effects of LNPs which will be i.v. injected in wild type mice (i.e. C57BL/6). In particular, we will determine i) acute toxicity: possible adverse effects after first exposure to a single dose of LNPs; ii) maximum tolerated dose (MTD): highest dose, in single dose regimes, showing no significant weight loss (<10%) and absence of adverse effects; iii) repeated-dose toxicity: identification whether toxicity occurs after continuous exposure to LNP. After treatments, blood samples will be collected to determine the levels of glutamate pyruvate transaminase (GPT), creatinine (CRE), and creatine kinase-MB (CK-MB). After mice are euthanized, liver, kidney and intestine will be subjected to biochemical and histological analyses (Xxxxxx et al, 2013).

Afterwards, we will test the capability of LNP-miRs to revert drug resistance in vivo using A375DR, LOX IMVIDR cell lines as well as two patients-derived cell lines (for a total of 4 models) injected in immunocompromised mice (i.e. CD1). We will set up combinatorial experiments of LNP-miRs with Dabrafenib (10 mg/kg/day) + Trametinib (0.3 mg/kg/day). In detail, the groups will be: a) LNP-SCR (control LNP), b) LNP-miRs, c) LNP-SCR + BRAFi + MEKi and d) LNP-miRs + BRAFi + MEKi. LNP-miRs will be dosed once, twice or three times per week in order to identifiy the most efficacious regimen to be translated to clinical trials. Treatments will be performed for 4 weeks and tumor growth will be measured twice a week untill mice are euthanized if tumor masses become bigger than 1500 mm3. Primary outcome will be the extent of inhibition of tumor growth. Tumor masses will be collected and analyzed to evaluate: a) apoptosis induction, proliferation, metastatization and tumor angiogenesis by IHC and b) xxXXX/target genes expression levels by Western Blot and qRT- PCR analyses, respectively.

Experimental design aim 3

Finally, we plan to further validate the LNP-miRs charachterized in the previous Aims as novel therapeutics in combination with MAPKi for melanoma in patient-derived xenografts (PDX) able to successfully recapitulate the tumor architecture and genotype of the original tumor. Tumor fragments will be implanted from melanoma patients directly into immunodeficient NOD-scid IL2Rgammanull (NSG) mice (Xxxxxx et al, 2016). To this aim, we plan to develop two different types of PDX models derived from BRAF mutated melanoma patients: 1) drug sensitive: before starting BRAFi and MEKi therapy, and 2) drug resistant: after progression from targeted therapy or immunotherapy (determined by RECIST criteria). Patients relapsing after treatments will be defined as patients who experienced progressive disease (PD) or stable disease (SD) for less than 6 months. Briefly, tumors will be harvested and PDX will be 1) bio-banked and cryopreserved, 2) subjected to transcriptomic (i.e. RNA-seq) and genomic (Whole Exome Sequencing [WES]) characterization, and 3) dissected for the re- implantation and propagation in additional NSG mice. These last mouse models will be subjected to pharmacokinetic- pharmacodynamic studies and efficacy studies.

The former (i.e. drug sensitive) will be initially utilized to determine the optimal LNP-miRs dosage in PDX models as previously done for standard xenografts, as well as the time frame of treatments in order to obtain a constant delivery to

tumors of the three miRNAs and the resulting inhibition of their relevant targets within the tumors. For this purpose we will extract RNA and proteins from PDX-derived tumor lesions. For these experiments, we will use PDX derived from drug naïve lesions that we expect to be more easily available as compared to those deriving from relapsing patients. Briefly, PDX will be implanted in mice and when tumors reach a volume of 150 mm3, mice will be i.v. injected with LNP-miRs or LNP-SCR and then euthanized at different time points following treatment (i.e., 24, 48, 72 and 96 hours). Tumor masses will be analyzed by qRT-PCR and Western Blot analyses to assess xxXXX/target genes expression levels. This set of experiments will enable the determination of the LNP-miRs doses and the time frame of treatments.

We then will move to efficacy studies which will be carried out according to standardized protocols established with stable cells lines described in Aim 2. The outcomes of the studies will be: 1) for PDX derived from drug sensitive naïve tumors, to demonstrate the therapeutic capability of LNP-miRs to potentiate targeted therapies, whereas 2) for PDX derived from drug resistant biopsies, to evaluate the therapeutic capability of LNP-miRs to restore drug sensitivity and to induce regression of tumor growth when target therapy is unable to do so. The frequency of LNP-MiRs delivery (once, twice or three times weekly will be decided on the basis of the results achieved with stable cell lines in CD1 mice as described in Aim 2. After mice sacrifice, tumor masses will be collected and analyzed by IHC, qRT-PCR and Western blot analyses as described above. Furthermore, we will also evaluate the effects of the treatments on the TME status given the capability of the three microRNAs to inhibit the expression of proinflammatory factors responsible for the maintenance of an immunosuppressive and drug resistant TME (see preliminary results). To this aim, abundance and characterization of tumor infiltrating immune cells will be assessed by multicolor FACS analyses using panels composed of multiple antibodies.

In conclusion the set of pre-clinical data obtained in the combined Aim2 and Aim3 will represent a robust package of data in support of future clinical studies in patients which have developed drug resistance. These data will be utilized either to apply to grants dedicated to clinical development or to look for biotech or industrial partners interested to support further this program

Picture to support preliminary data

Picture to support Preliminary Data_PoC Xxxxxxxxx.pdf

Hypothesis and significance

This proof-of-concept project stems from two robust set of data: a) the identification (and patenting) of a group of oncosuppressor microRNAs involved in the establishment of drug resistance against targeted therapy in BRAF-mutated melanoma ((Xxxxxxx et al 2018; PCT/IT2019/050073: miRNAs for treatment and diagnosis of drug resistant tumors); b) the more recent demonstration, both in vitro and in xenograft models, that LNPs delivered miRNAs potentiate the antitumor efficacy of targeted therapy in drug sensitive BRAF-mutated cell lines (Fattore et al, manuscript under revision). Preliminary results:

1. MAPKi-resistant melanoma cells down-regulate two oncosuppressive miRNAs, namely miR-204-5p and miR-199b-5p as compared to sensitive counterparts. The same cells in turn overproduce and release a wide repertoire of pro-angiogenic and pro-inflammatory factors (e.g. VEGF, TGFß1, CCL5 and CXCL2), which are molecular target of the above mentioned miRNAs (Figure 1).

2. LNPs-carrying oncosuppressive miRNAs (miR-199b-5p, miR-204-5p) efficiently deliver their therapeutic cargo and inhibit the pro-angiogenic and pro-inflammatory factors released by drug resistant melanoma cells. Furthermore, these therapeutics are able to reduce the proliferation of BRAFi resistant A375 and M14 melanoma cells in the presence of Dabrafenib+Trametinib (i.e. MAPKi) (Figure 2).

3. MAPKi resistant melanoma cells recruit and reprogram tumor associated macrophages (TAMs). Oncosuppressor miRNAs (miR-199b-5p, miR-204-5p) delivered by LNPs re-educate M2 polarized macrophages instructed by drug resistant melanoma cells (Figure 3).

4. LNP-carrying miR-204-5p and miR-199b-5p (40 ¿g) was able to reduce tumor masses of A375 xenograft melanoma

model as compared to LNP-Scr (control) treated mice. At 72 hours post-injection, we also observed the highest level of overexpression of both miRNAs coupled with the highest degree of reduction of their mRNA targets within tumor samples (Figure 4).

5. In vivo delivery of LNP-miR (miR-204-5p and miR-199b-5p) strongly potentiates MAPKi therapy in two different melanoma xenograft models (i.e. A375 and M14) and achieves a longer disease control as compared to the sole MAPKi while depleting TAMs infiltrating in tumors (Figure 5 and 6).

6. Triple regimen LNP-miR+BRAFi+MEKi was well tolerated by A375 and M14 xenograft models: any significant weight loss in mice as compared to the beginning of the study in both the xenograft models was not observed (Figure 7).

7. The characteristics of LNP-Scr or encapsulating miR-204-5p/199b-5p are reported in Table 1.

However, in order to advance to a clinical trial, we need the following additional pre-clinical evidence: a) define a final, reproducible, easy to formulate and easy scalable production of oncosuppressor miRNAs loaded LNPs; b) execute preliminary toxicology studies demonstrating optimal biodistribution, elimination and tolerability of LNP-miRs; c) demonstrate efficacy of these LNP-miRs in a variety of drug resistant mouse models, in the best case PDX mice derived from tumor lesions of drug resistant patients.

We hypothesize that the efforts of our joint team will allow us to obtain these results in the two-year time frame of the project.

The significance of our findings is highly translational towards clinical development. This project is a classical example of how initial findings obtained from the study of patient's samples generates a hypothesis that could be transfgerred to the bedside in the same type of patients.

5.5 Methodologies and statistical analyses

Methods of data collection

Researchers/oncologists in charge of the study will assign an identification code to each sample/mice/patient. They will be the only investigators able to trace the identity of the sample/mice or people enrolled/recruited. The data will be stored in a dedicated database and used only by authorized personnel. This will guarantee quality, integrity, security, availability and traceability, under the responsibility of the workforce in charge of the study. We will make sure that the whole study will be carried out in full compliance with the requirements of the legislation for the processing of personal data and guaranteed by the reference legislation in force. All the data, documentation and material of the studies carried out will be kept by the people in charge of the study for a period not exceeding that necessary to achieve the scientific research purposes of the study. This will be established by the legislation and regulations governing the matter, in a manner that guarantees its confidentiality (for a period of at least 5 years from the conclusion of the study or for a longer period). Access to computer systems where data are stored/processed, will be controlled by means of appropriate security measures and by authorized personnel. The people in charge of the study will adopt appropriate technical and organizational measures to guarantee an adequate level of security of the data (and/or samples) processed in the context of the study, as well as specific measures and technical measures to increase the security and to prevent the dissemination of personal data or their use by unauthorized parties. Patients will authorize the use of their samples by signing a general informed consent to allow the analysis of material for research purposes which will be tested in an anonymous manner.

Statistic plan

Aim1) For each formulation, three small-sized batches will be prepared and characterized in terms of colloidal properties and miR encapsulation efficiencies; the results will be expressed as average ±SD across the different batches. The nanoparticle stability against aggregation in serum and the hemolysis assays will be carried out on three independent batches with two technical replicates for each batch.

Aim 2) In vitro experiments will be replicated at least three times and the data will be expressed as average ±SD or ±SE of the mean (SEM). We will consider 4 BRAF mutated cell lines after development of drug resistance. To determine

differentially expressed miRNAs/genes we will use paired Student's t-test applied to log2 transformed data. For the in vivo setting, we will use groups of 7 mice (about 6 weeks old) and all the experiments will be repeated at least twice. In particular, to test toxicity, immunogenicity and side effects of LNPs we will use C57BL/6 mice for a total number of 30. For drug efficacy studies we will use the total number of 188 CD1 nude mice (7 mice for each of the six experimental groups for four cell lines, see aim 2 = 168 plus additional 20 to ensure adequate randomizazion of tumor masses growth before initioation of drug dosing). The sample size has been calculated by using Gpower 3.1 as sufficient to have a statistical power (1-ß) > 80% and a significant level of 5%. For ex vivo experiments, we will consider a number of 3 patients. Thius was determined on the basis of ethical considerations about biopsy feasibility which will be possible only in patients with easily accessible skin metastases. This number was calculated to be sufficient to test an effect size equal or greater than

0.90 at a significance level of 5% with a power of 80%.

Aim 3) For PDX models we plan to use a total 190 NSG mice. 30 NSG mice will be utilized for initial expansion of 2 PDX from drug sensitive lesions (n=15) and 2 PDX from drug resistant lesions (n=15). 30 NSG mice will be utilized for phamacokinetic and pharmacodynamic studies. Finally 130 NSG mice will be used for efficacy studies (4 PDX mouse models - 2 sensitive 2 resistant, 4 groups of 7 animals per protocol, 28 + 28 =56 x 2 = 112 plus 18 mice to ensure randomization of tumor masses (experimental details are the same described for stable cells). The sample size has been calculated by using Gpower 3.1 as previously reported.

Statistical analysis

For LNP formulations, three small-sized batches will be prepared and characterized in terms of colloidal properties and miR encapsulation efficiencies; the results will be expressed as average ±SD across the different batches. In vitro experiments will be replicated at least three times and the data will be expressed as average ±SD or ±SE of the mean (SEM). Statistical analyses will be performed using GraphPad Prism v8.0 software. In vitro and in vivo groups will be compared by Student's t test or Xxxxxxxx Signed Rank Sum Test as indicated, and statistical significance will be represented as follows: *p < 0.05;

**p < 0.01; and ***p < 0.001. For identifying differentially expressed genes (DEGs) from RNA-seq data, a filter cut-off criterion of | log2FC|> 0.15 will be applied and genes with an adjusted p-value<0.05 will be considered as statistically significant. The biological function of DEGs will be identified by a Gene Ontology (GO) analysis using the R package ''enrichR'' (Xxxxxxxx et al, 2016).

The criteria for the selection of the miRNAs/genes deregulated in MAPKi-resistant cells will be a significant t-test (P<0.05) and an at least two-fold change from baseline data. For in vivo studies statistical analysis will be performed using GraphPad Prism v8.0 software by applying Two-sided T-Test and conducting a correction for multiple comparisons. Biostatistics and Bioinformatics Unit of our Institute will give support to the analyses of the results obtained throughout the project.

Timing of analysis data

Aim 1) This will be carried out throughout the first year; more specifically, months 1-5 will be dedicated to the production and characterization of LNP-miRs with the standard ethanol injection method while months 5-12 will be dedicated to the optimization of the microfluidic mixing method and the TFF purification protocols.

Aim 2) The results of the in vitro experiments will be analyzed throughout the first year of the project. The selection of MAPKi resistant melanoma cells will last about two months. For C57BL/6 models, to evaluate the potential toxicity, immunogenicity and side effects of LNPs, treatments will last about 7 days and then samples will be collected for the analyses of plasma and organs. For xenograft models, based on our previous experience (see preliminary results), treatments will start about 7-10 days after cell injection, depending on the cell lines. Mice will be euthanized when tumor volume reaches 1000 mm3 and treatments will be performed for 28 days. For combinatorial experiments, mice will be treated with LNPs (40 ug every 24 or 48 or 72h i.v.) and/or Dabrafenib (10 mg/kg/day) + Trametinib (0.3 mg/kg/day) (5 days/week o.g).

Aim 3) PDX models will be set up during the first year of the project and pharmacology/efficacy studies will be carried out

during the second year. Tumor fragments from melanoma patients will be implanted subcutaneously into the flanks of NSG mice for an initial propagation until tumor masses reach about 000-0000 mm3 in size. Then, tumors will be harvested and 1) bio-banked and cryopreserved, 2) subjected to RNA-seq and Whole Exome Sequencing and 3) dissected for the re- implantation and propagation in additional mice. Pharmacokinetic-pharmacodynamic studies will be performed injecting PDX-bearing mice with LNP-miRs or LNP-SCR via tail vein at the dose of 20 microg or 40 microg. Mice will be then euthanized at different time points following treatment (i.e., 24, 48, 72 and 96 hours) for the analyses described in experimental design 2. Combinatorial experiments will be performed according to protocols established with stable cells lines described above.

5.6 Expected outcomes

Our project is expected to provide a breakthrough in the management of melanoma patients maximizing patients' life expectancy and quality of life through a combinatorial approach. To reach our goal we will make us of oncosuppressor miRNAs encapsulated in LNPs, that represent a promising approach for therapy relying on their ability to i) target multiple oncogenic pathways relevant for melanoma progression and/or adopted by melanoma cells to survive upon treatment with target therapy, ii) circumvent the main drawbacks of delivering naked RNAs, i.e. poor cellular uptake, off-target activity and nuclease degradation. We expect that this project could have a relevant impact for the NHS, in terms of increased chance of cure and overcoming of drug resistance. Patients who, for various reasons, cannot benefit from currently available therapies or who develop resistance after initial treatment could benefit from new therapeutic strategies identified through this project.

5.7 Risk analysis, possible problems and solutions

As most of the techniques required to reach the proposed aims are performed on a regular basis, the team does not foresee substantial issues in carrying out the planned experiments. If necessary the team has the ability to carry out additional approaches as outlined below:

1) in addition to the LNPs formulation described in specific aim 1, we will be able to set up and develop additional formulations depending on the results that will be obtained;

") a peptide functionalization strategy may be envisaged to enhance the in vivo targeting ability of LNPs. Specifically, the LNP surface may be decorated with an alpha melanocyte stimulating hormone (e.g. alphaMSH) peptide able to bind to the melanocortin-1 receptor (MC1-R), which is overexpressed on the surface of human melanoma cells. The alphaMSH peptide will be functionalized with a thiol moiety for conjugation to a maleimide-terminated PEGylated lipid, which will be incorporated within the LNP composition (Xxxxxxxxxx et al 2013, Xxxxx et al 2020).

5.8 Significance and Innovation

Current research is exploring the role of combinatorial approaches to overcome drug resistance, frequently observed in melanoma. Several clinical trials are ongoing to evaluate the efficacy of target therapy in combination with immunotherapy or drugs with different mechanism of action but, to the best of our knowledge, at present no trials are studying the possibility of using miRNAs to synergise with target therapy. With this project, our group aims to evaluate this possibility with the final goal to identify new therapeutic approaches for the treatment of metastatic melanoma patients. Therapeutically, the transition of LNPs-xxXXX-based therapies to the clinical practice in melanoma is an innovation and will be facilitated by the success in the fight against the SARS-CoV2 pandemic of LNPs-based vaccines. The market value of our approach will increase the probability to obtain additional financial support to conduct a Phase I/II clinical trial in the following years

5.9 Bibliography

- Xxxx XX, et al. Nat Rev Clin Oncol. doi: 10.1038/nrclinonc.2017.43.

- Xxxxxx X, et al. N Engl J Med. doi: 10.1056/NEJMoa1904059

- Xxxx L, et al. Nat Cancer. doi: 10.1038/s43018-021-00221-9

- Fattore L, et al. Cell Death Differ. doi: 10.1038/s41418-018-0205-5.

- Fattore L, et al. Int J Mol Sci. doi: 10.3390/ijms21061930.

- Campani V et al. International Journal of Pharmaceutics. doi: 10.1016/j.ijpharm.2020.119693

- Pramanik et al. Mol Cancer Ther. doi:10.1158/1535-7163.MCT-11-015

- Veiseh et al. Mol Pharm. doi: 10.1021/mp4005468.

- Lo et al. Theranostics. doi: 10.7150/thno.45164

- Kruiswijk et al, Oncogene. doi:10.1038/onc.2015.282

- Xxxxxx et al, Clin Cancer Res. doi:10.1158/1078-0432.CCR-15-1762.

- Gogas H, et al. J Xxxx Xxxxx 0000; 38: 10012.

- Xxxxxxx et al, Int J Mol Sci. doi: 10.3390/ijms22083837.

- Xxxxxx et al, Molecular Therapy Nucleic Acids. doi: 10.1016/j.omtn.2019.10.045

- Rosenkranz et al. Biochem. doi: 10.1134/X0000000000000000.

- Xxxxx, X. et al. Biomaterials. doi: 10.1016/j.biomaterials.2020.119858

5.10 Timeline / Deliverables / Payable Milestones

D1.1: Formulation/characterization of LNP-miRs (Months 1-5)

M1.1: production of LNP-miR with desired hydrodynamic/encapsulation efficiency D2.1: Formulations via microfluidic mixing (Months 5-10)

M2.1: LNP-miR formulations obtained by microfluidic mixing

D3.1: Purification of LNP-miRs via tangential flow filtration (Months 9-12) M3.1: establishment of a flow filtration protocol

D1.2: In vitro models (Months 1-6)

M1.2: Assess miRs/targets and cell viability upon LNP-miRs D2.2: Ex vivo models (Months 7-12)

M2.2: melanoma biopsies incubated with LNP-miRs to test their efficacy D3.2: In vivo models (Months 7-20)

M3.2: Test LNPs toxicity, drug efficacy studies, analyses of tumors

D1.3: PDX collection/characterization (Months 1-15). M1.3: PDX bio-banked, sequenced and propagated.

D2.3: pharmacokinetic-pharmacodynamic studies (Months 7-12). M2.3: determine LNPs dosage in PDX

D3.3: drug efficacy studies in PDX (Months 13-24).

M3.3: combo of LNP-miRs + MAPKi in PDX; analyses of tumors

Milestones 12 month

At 12 month from the study set up: a) the formulation and characterization of LNP-miRs will be completed; b) LNP-miRs

efficacy in combination with target therapy will be determined in several drug resistant melanoma preclinical models; c) PDX collection will be established and tested for pharmacokinetic-pharmacodynamic studies

Milestones 24 month

After 24 month, in vivo studies on PDX models will be completed, along with the extensive molecular characterization of all the tumor masses from mice collected throughout the project. Data will be analyzed and results interpreted in light of the hypothesis stated. Scientific dissemination of the study results will be pursued in form of communications and scientific manuscripts.

Gantt chart

gantt_chart PoC Xxxxxxxxx .xlsb

5.11 Equipment and resources available

Facilities Available

The Department of Pharmacy of the University of Xxxxxx Xxxxxxxx XX carries out training activities and basic and applied research towards the discovery of new drug targets and innovative therapeutic strategies. The Department of Pharmacy has been named among the Departments of Excellence for the years 2018-2022 by the Ministry of University and Research. The lab of Prof. De Rosa has all the necessary equipment to carry out the project; in particular, it is equipped with a Xxxxxxx Zeta Sizer Ultra instrument for the characterization of colloidal size and surface charge of nanoparticles, a Xxxxxxx Xxxxxxx ultracentrifuge for the purification of formulations, a Shimadzu spectrophotometer for the quantification of xxXXX, and a differential scanning calorimeter (DSC) to characterize the thermal behavior of the formulations. IRCCS Regina Elena National Cancer Institute (IRE) in Rome and IRCCS Istituto Xxxxxxx National Cancer Center in Napoli are both OECI Accredited Comprehensive Cancer Centers. They act as referral centers in central and south Italy respectively. Clinical units work closely with the advanced research laboratories. IRE is furnished of all the facilities and equipment necessary for the project. Proteomics, genomics, sequencing apparatus, FACStar, FACS SCAN, IVIS® Spectrum system for imaging and image processing of luminescent and fluorescent samples, confocal and immunofluorescence microscopies, ChIP-on-ChIP, Snc-RNAs, proteomics and Affimetrix technologies are available. The Institute has access to an external qualified Animal Facility. In addition, the Institute takes advantage of Biostatistical and Bioinformatic Unit for the statistical analysis of clinical and laboratory data. Finally, IRE Biobank is a reliable resource of human samples for cancer research. At INT-Xxxxxxx are present biohazard flow hoods; centrifuges and microscopes dedicated to cell and tissue cultures; liquid nitrogen frozen apparatus; several apparatus for RealTime PCR; ELISAplate reader; Fluorescence Multilabel Reader; Flow Aspirated Cell Scan (FACScalibur). Two Animal Care Facilities are also available with several imaging systems: the IVIS Lumina XR to follow tumor progression, monitor response to therapeutic treatments, metastasis, cell tracking, and tumor burden monitoring; SKYSCAN 1276 microCT system for high resolution, preclinical in vivo and ex vivo studies, the Leica MacroFluo system macroscope for ex vivo and in vivo fluorescence image analysis.

Subcontract

Not applicable

5.12 Desc. of the complementarity and sinergy of secondary collab. researchers

The most important feature of this proposal is the complementarity of the research team which covers in vitro, in vivo, ex vivo and clinical expertise required to successfully reach the objectives. Scientists will be matched by clinicians, not only for the primary sample collection, but also in a spirit of collaboration and information exchange flow, from bench to bedside and vice versa. The expertise of Xxxx. Xxxxxxxxx goes from biology of lung cancer stem cells to immunotherapy and generation of tumor vaccines and the relevance of intestinal microbiota in immunogenicity. In the last years he contributed to the study of COVID19 in oncological patients, and identified a panel of miRNAs as possible therapeutics for melanoma treatment. Dr.

Xxx Xxxxxx has a long lasting experience in the in vitro and in vivo studies on melanoma progression and response to

therapy. As co-PI she will monitor the proper advancement of the project, together with the PI. Xxxx. Xxxxxxx is an expert in the field of new biomarkers in cancer stem cell derived by malignant pleural effusion and melanoma, and in the identification of possible pathways involved in the resistance to BRAF/MEK inhibitors in melanoma. Dr. Ascierto is an expert oncologist clinician not only nationally but also internationally renowned. His major research interests have included research in the genetics and proteomics of melanoma, assessment of new molecular markers for tumor progression, biochemical and immunological monitoring, immunotherapy, target therapy, combination strategies and vaccination treatments in solid tumors, with particular regard to melanoma. He will be in charge of melanoma patient recruitment and follow up. Prof. Xx Xxxx'x scientific activity is focused on the design and development of vehicle systems based on micro and nanotechnologies and in particular on lipid nanoparticles for the in vitro and in vivo delivery of therapeutic miRNAs. Dr. Xxxxxxxxxx is an expert in system biology and computational and bioinformatics approaches for the study of biological activity of proteins. More recently she focused her attention on inflammation and molecules involved in this phenomenon.

Dr. Fattore and Dr. Di Martile are young researchers who focused their attention, respectively to miRNAs and bcl-2 family members in melanoma pathobiology. Both of them show documented expertise to perform most of the in vitro/in vivo experiments required for the project.

Several papers published by the PI in collaboration with all components of the team support the already existing collaboration in the proposal field (Xxxxxxxxxx S et al 2022; Tupone MG et al 2020; Xxxxxx ME et al 2018; Xxxxxxx L et al 2016; Xxxxxxxx N et al 2015; Xxxxxxx et al Int J Mol Sci 2020). The project development will also take advantage from internal collaborators, as well as, pathologists and surgeons with experience in the management of melanoma, and bioinformaticians for statistical analyses. To further support the synergy of research team, three members of this group have patented the use of a subset of miRNAs in combination with targeted therapy to revert drug resistance and the systemic delivery of oncosuppressor miRNAs using LNPs (PCT/IT2019/050073). Finally, the four Institutions have co- authored a manuscript currently under review ("Unravelling a complex regulatory network involved in the development of an immunosuppressive tumor microenvironment and drug resistance in melanoma")

5.13 Translational relevance and impact for the national health system (SSN)

What is already know about this topic?

Metastatic melanoma, a highly aggressive tumor, can be fatal within 18 months of diagnosis (Xxxxxxx et al, 2021). Targeted therapies with inhibitors of MAPK pathway have greatly improved overall survival in about 50% of patients with BRAF-V600 mutations (Xxxxxx et al, 2019). However, drug resistance limits their long-term efficacy (Xxxxx et al, 2020). miRNAs have emerged as orchestrators of this phenomenon in melanoma (ref). The PI has explored their therapeutic applications (Xxxxxxx et al, 2018). In this context, LNPs are promising nanovectors for xxXXX delivery (Xxxxxx et al, 2020). The approval of Onpattro® (LNPs encapsulating siRNA) and the SARS-CoV2 pandemic outbreak have dramatically accelerated the clinical adoption and scale-up manufacturing of nucleic acid-loaded LNPs. Finally, several members of our team have shown the ability of LNPs carrying oncosuppressive miRNAs to reduce cell growth alone and, more efficiently, when combined with MAPKi (Xxxxxxx et al, 2020).

Details on what is already know about this topic

The rapid development of knowledge of miRNAs regulation suggests that they can become therapeutic targets or they may become drugs for a broad spectrum of pathologies, including cancer. Indeed, a relevant concept is that a xxXXX can regulate expression of multiple target proteins, interacting with multiple target mRNAs. Every year in Italy, about 7300 new diagnosed cases of melanoma are estimated, and about 1500 deaths are recorded, most of them due to drug resistance, that occurs after treatment with targeted drugs. The PI has recently patented the systemic delivery of oncosuppressor miRNAs using LNPs for melanoma. Among others miR-204-5p and miR-199b-5p exert a direct antitumor effect and xxxxx the development of drug resistance Hence, the use of these miRNAs could represent a new modality of "epigenetic therapy" that could be applied alone or in combination with already available therapies, to improve the current treatment of melanoma.

What this reasearch adds?

Our project has important implications for cancer patients because it is directed to identify and validate, in a series of in vitro and in vivo models, new targets for combination therapies in metastatic melanoma. To our knowledge there are no published studies, and not even clinical trials, that are investigating the usefulness of xxXXX-loaded LNPs to potentiate targeted therapies for BRAF-mutated melanomas. This is what our proof-of-concept project will pursue. We believe that our results will support our purpose, demonstrating that LNPs encapsulating oncosuppressive miRNAs are novel therapeutics capable of simultaneously hitting many intrinsic and extrinsic oncogenic avenues adopted by melanoma cells to survive upon MAPK pathway inhibition. Furthermore, we plan to lay the groundwork for the easy scalable production of xxXXX- loaded LNPs paving the way for the future clinical translation and industrial scale-up of these formulations to design Phase

I-II studies.

Details on what this reasearch adds

Our project aims at providing new insights into an area at the forefront of molecular oncology: to identify new therapeutic strategies for melanoma patients who develop resistance to therapy. As resistance may be due to several mechanisms including miRNAs, our proposal aims to study the possibility of using miRNAs to potentiate currently available therapies. Our findings and discoveries could be exploited by biotech or pharma companies, and serve as robust preclinical data for designing new clinical trials, especially for the subset of melanoma patients carrying BRAF mutations and developing resistance to target therapy. In addition to the numerous clinical trials evaluating the role of miRNAs expression in blood as potential diagnostic and prognostic biomarkers or for stage stratification for cancer patients, our results could allow the recruitment of melanoma patients for new clinical trials evaluating the potential efficacy of therapeutic miRNAs in combination with target therapy.

What are the implications for public health, clinical practice, patient care?

Acquired resistance to targeted therapies in BRAF-mutated metastatic melanomas generates in many cases an intractable clinical condition. Patients relapsing after treatments with MAPKi have lower response rate to immunotherapies with ICI (Xxxx et al, 2021). From here, the need to develop novel therapeutics able to a) prolong the efficacy of targeted therapies and b) to restore drug sensitivity. Supported by our previously published and preliminary data, this proposal addresses a highly unmet medical need. The translational relevance of this project for public health, clinical practice relies on the possibility to set up an easy scalable protocol for the production of xxXXX-loaded LNPs, which will be also tested for the preliminary toxicology to determine their in vivo bio-distribution and tolerability. The outcomes of these translational studies pave the way for the future clinical trials in the attempt to propose RNA-based therapeutics as novel tools for melanoma patient's care.

Details on what are the implications for public health, clinical practice, patient care

The research described in this proposal is expected to generate results with both short- and long-term impacts on the clinical management of melanoma. It could lead to the identification of new therapies to improve the clinical outcome for patients with melanoma, to enhance survival and to improve quality of life. In this context, the expression of some miRNAs in melanoma represents a key element that affects the progression of melanoma and the resistance to available drugs.

RNA therapies (therapeutic miRNAs or small interfering RNAs) represent some of the most important biopharmaceuticals for future medicines encompassing a rapidly expanding drug category that will change the standard of care for many diseases and modernize personalized medicine. Through the modulation of some miRNAs in combination with treatments currently in use for the therapy of melanoma, our project could produce encouraging experimental results in support of new therapeutic approaches to be translated into clini

6 - Budget

Total proposed budget ( Euro )
Costs	TOTAL BUDGET	Co-Funding	List of costs proposed for funding to the MOH	Percentage of total proposed to the MOH
1 Staff Salary	265.800,00	265.800,00	not permitted	0,00
2 Researchers' Contracts	250.000,00	0,00	250.000,00	26,04
3a.1 Equipment (Leasing -	0,00	0,00	0,00	0,00
3a.2 Equipment (buying)	0,00	0,00	0,00	0,00
3b Supplies	538.800,00	0,00	538.800,00	56,12
3c Model Costs	55.000,00	0,00	55.000,00	5,73
4 Subcontracts *	0,00	0,00	0,00	0,00
5 Patient Costs	0,00	0,00	0,00	0,00
6 IT Services and Data Bases	0,00	0,00	0,00	0,00
7 Travels	16.000,00	0,00	16.000,00	1,67
8 Publication Costs	20.000,00	0,00	20.000,00	2,08
9 Dissemination	8.000,00	0,00	8.000,00	0,83
10 Overheads *	67.200,00	0,00	67.200,00	7,00
11 Coordination Costs	5.000,00	0,00	5.000,00	0,52
Total	1.225.800,00	265.800,00	960.000,00	100,00

* percentage calculated as average value between all the Operating Units.

Report the Co-Funding Contributor:

For Unit 1 95.000 in total: 50.000 € calculated as 2 months/man each of 2 senior scientists (Xxxxxxxxx and Xxx Xxxxxx) + 40.000€ for 6 months/man each of two under 40 Fattore and Di Martile

For Unit 2 40.800 € calculated as 5 months/man in total for senior researcher xxxx. Xxxxxxx For Unit 3 50.000 € calculated as 4 months/man in total for senior researcher prof. De Rosa

For Unit 4 80.000 € in total: 50.000 € calculated as 4 months/man for senior clinician Dr. P.A. Ascierto + 30.000 as 6 months/man for Senior Staff Researcher Dr Xxxxxxxxxx

Budget Justification
1 Staff Salary	Co-funding with man months of staff personnel salaries as detailed inm the budget section
2 Researchers' Contracts	Fellowships for 5 new young U40 fellows to be hired for the 2 years duration of the project of which 3 in South Italy (2 at Xxxxxxx Institute and 1 at Univ Xxxxxxxx XX in Napoli) calculated at the cost of 25.000€/fellowship/year

3a.1 Equipment (Leasing - Rent)	Not Applicable
3a.2 Equipment (buying)	Not Applicable
3b Supplies	Chemicals for nanoparticles generation. Reagents for cell cultures, Cytoplex , RNA-seq analysis. kits for RNA and xxXXX extraction, probes for qRT-PCR. Antibodies and kits for flow cytometry and IHC. Drugs for in vitro and in vivo studies (Dab and Tram)
3c Model Costs	Purchase of C57Bl/6, CD1 and NSG mice as detailed in the relevant project's sections, costs for mice transport, handling and feeding
4 Subcontracts	Not Applicable
5 Patient Costs	Not Applicable
6 IT Services and Data Bases	Not Applicable
7 Travels	Calculated based in the participation of 8 members of the team to one national or international (european) congress with an average cost of 1000 € per congress per person
8 Publication Costs	Submission and publication costs of manuscripts on international peer-reviewed open access journals. Calculated as 2500€ average per publication, two publications per unit
9 Dissemination	Registration fees for national or international meetings, and for printing posters, as the results obtained require dissemination among the scientific community and the general public. Calculated as 2000€ average per Unit
10 Overheads	Calculated as flat 7% for all consortium members
11 Coordination Costs	Calculated as 2 alignment meetings of the consortium members one per year of the duration of 2 days with invitation of 2 external advisors at the cost of 2500€/meeting

Proposed total budget UO1 Institution: Istituti fisioterapici ospitalieri - Istituto Regina Elena (Euro)

Costs	TOTAL BUDGET	Co-Funding	List of costs proposed for funding to the MOH	Percentage of total proposed to the MOH
1 Staff Salary	95.000,00	95.000,00	not permitted	0,00
2 Researchers' Contracts	50.000,00	0,00	50.000,00	20,00
3a.1 Equipment (Leasing - Rent)	0,00	0,00	0,00	0,00
3a.2 Equipment (buying)	0,00	0,00	0,00	0,00
3b Supplies	146.500,00	0,00	146.500,00	58,60
3c Model Costs	20.000,00	0,00	20.000,00	8,00
4 Subcontracts	0,00	0,00	0,00	0,00
5 Patient Costs	0,00	0,00	0,00	0,00
6 IT Services and Data Bases	0,00	0,00	0,00	0,00
7 Travels	4.000,00	0,00	4.000,00	1,60
8 Publication Costs	5.000,00	0,00	5.000,00	2,00
9 Dissemination	2.000,00	0,00	2.000,00	0,80
10 Overheads	17.500,00	0,00	17.500,00	7,00
11 Coordination Costs	5.000,00	0,00	5.000,00	2,00
Total	345.000,00	95.000,00	250.000,00	100,00

Budget Justification
1 Staff Salary	Co-funding with man months of staff personnel salaries as detailed in the budget section
2 Researchers' Contracts	Fellowship for two years for one new U40 biologist calculated at the cost of 25.000€ per year
3a.1 Equipment (Leasing - Rent)	N/A
3a.2 Equipment (buying)	N/A
3b Supplies	Reagents for cell cultures, Cytoplex , RNA-seq analysis. kits for RNA and xxXXX extraction, probes for qRT- PCR. Drugs for in vitro and in vivo studies (Dab and Tram)
3c Model Costs	Purchase of C57Bl/6 and CD1 mice as detailed in the relevant project's sections, costs for mice transport, handling and feeding
4 Subcontracts	N/A
5 Patient Costs	N/A
6 IT Services and Data Bases	N/A
7 Travels	Calculated based on the participation of 2 members of the team to one national or international (european) congress with an average cost of 1000 € per congress per person
8 Publication Costs	Submission and publication costs of manuscripts on international peer-reviewed open access journals. Calculated as 2500€ average per publication, two publications
9 Dissemination	Registration fees for national or international meetings, and for printing posters, as the results obtained require dissemination among the scientific community and the general public. Calculated as 2000€ average
10 Overheads	Calculated as flat 7%
11 Coordination Costs	Calculated as 2 alignment meetings of the consortium members one per year of the duration of 2 days with invitation of 2 external advisors at the cost of 2500€/meeting

Proposed total budget UO2 Institution: Sapienza - University of Rome (Euro)

Costs	TOTAL BUDGET	Co-Funding	List of costs proposed for funding to the MOH	Percentage of total proposed to the MOH
1 Staff Salary	40.800,00	40.800,00	not permitted	0,00
2 Researchers' Contracts	50.000,00	0,00	50.000,00	20,83
3a.1 Equipment (Leasing - Rent)	0,00	0,00	0,00	0,00
3a.2 Equipment (buying)	0,00	0,00	0,00	0,00
3b Supplies	162.200,00	0,00	162.200,00	67,58
3c Model Costs	0,00	0,00	0,00	0,00
4 Subcontracts	0,00	0,00	0,00	0,00
5 Patient Costs	0,00	0,00	0,00	0,00
6 IT Services and Data Bases	0,00	0,00	0,00	0,00
7 Travels	4.000,00	0,00	4.000,00	1,67
8 Publication Costs	5.000,00	0,00	5.000,00	2,08
9 Dissemination	2.000,00	0,00	2.000,00	0,83
10 Overheads	16.800,00	0,00	16.800,00	7,00
11 Coordination Costs	not permitted	not permitted	not permitted	0,00
Total	280.800,00	40.800,00	240.000,00	100,00

Budget Justification
1 Staff Salary	Co-funding with man months of staff personnel salaries as detailed in the budget section
2 Researchers' Contracts	Fellowship for one new young U40 fellow to be hired for the 2 years duration of the project calculated at the cost €25.000 per year
3a.1 Equipment (Leasing - Rent)	N/A
3a.2 Equipment (buying)	N/A
3b Supplies	Reagents for cell culture (culture medium, antibiotics, fetal bovine serum), kits for RNA and xxXXX extraction, probes for qRT-PCR. Antibodies and kits for flow cytometry and IHC. Plastics/glassware (tubes, petri dishes,pipettes, flasks, multiwells).
3c Model Costs	N/A
4 Subcontracts	N/A
5 Patient Costs	N/A
6 IT Services and Data Bases	N/A
7 Travels	Calculated based in the participation of 2 members of the team to one national or international (european) congress with an average cost of 1000 € per congress per person
8 Publication Costs	Submission and publication costs of manuscripts on international peer-reviewed open access journals. Calculated as 2500€ average per publication, two publication
9 Dissemination	Registration fees for national or international meetings, and for printing posters, as the results obtained require dissemination among the scientific community and the general public. Calculated as 2000€ average
10 Overheads	Calculated as flat 7%
11 Coordination Costs	N/A

Proposed total budget UO3 Institution: Università degli Studi di Xxxxxx Xxxxxxxx XX (Euro)

Costs	TOTAL BUDGET	Co-Funding	List of costs proposed for funding to the MOH	Percentage of total proposed to the MOH
1 Staff Salary	50.000,00	50.000,00	not permitted	0,00
2 Researchers' Contracts	50.000,00	0,00	50.000,00	21,28
3a.1 Equipment (Leasing - Rent)	0,00	0,00	0,00	0,00
3a.2 Equipment (buying)	0,00	0,00	0,00	0,00
3b Supplies	157.550,00	0,00	157.550,00	67,04
3c Model Costs	0,00	0,00	0,00	0,00
4 Subcontracts	0,00	0,00	0,00	0,00
5 Patient Costs	0,00	0,00	0,00	0,00
6 IT Services and Data Bases	0,00	0,00	0,00	0,00
7 Travels	4.000,00	0,00	4.000,00	1,70
8 Publication Costs	5.000,00	0,00	5.000,00	2,13
9 Dissemination	2.000,00	0,00	2.000,00	0,85
10 Overheads	16.450,00	0,00	16.450,00	7,00
11 Coordination Costs	not permitted	not permitted	not permitted	0,00
Total	285.000,00	50.000,00	235.000,00	100,00

Budget Justification
1 Staff Salary	Co-funding with man months of staff personnel salaries as detailed in the budget section
2 Researchers' Contracts	Fellowship for 1 new young U40 fellow to be hired for the 2 years duration of the project calculated at the cost of 25.000€ per year
3a.1 Equipment (Leasing - Rent)	N/A
3a.2 Equipment (buying)	N/A
3b Supplies	Ionizable lipids, neutral lipids and PEGylated lipids; reagents for laboratory analysis, solvents, plastic and laboratory glassware, consumables for dimensional and zeta potential analysis, consumables for HPLC.
3c Model Costs	N/A
4 Subcontracts	N/A
5 Patient Costs	N/A
6 IT Services and Data Bases	N/A
7 Travels	Calculated based in the participation of members of the team to one national or international (european) congress with an average cost of 1000 € per congress per person
8 Publication Costs	Submission and publication costs of manuscripts on international peer-reviewed open access journals. Calculated as 2500€ average per publication, two publications
9 Dissemination	Registration fees for national or international meetings, and for printing posters, as the results obtained require dissemination among the scientific community and the general public. Calculated as 2000€ average
10 Overheads	Calculated as flat 7%
11 Coordination Costs	N/A

Proposed total budget UO4 Institution: IRCCS Istituto Nazionale Tumori Fondazione X. Xxxxxxx (Euro)

Costs	TOTAL BUDGET	Co-Funding	List of costs proposed for funding to the MOH	Percentage of total proposed to the MOH
1 Staff Salary	80.000,00	80.000,00	not permitted	0,00
2 Researchers' Contracts	100.000,00	0,00	100.000,00	42,55
3a.1 Equipment (Leasing - Rent)	0,00	0,00	0,00	0,00
3a.2 Equipment (buying)	0,00	0,00	0,00	0,00
3b Supplies	72.550,00	0,00	72.550,00	30,87
3c Model Costs	35.000,00	0,00	35.000,00	14,89
4 Subcontracts	0,00	0,00	0,00	0,00
5 Patient Costs	0,00	0,00	0,00	0,00
6 IT Services and Data Bases	0,00	0,00	0,00	0,00
7 Travels	4.000,00	0,00	4.000,00	1,70
8 Publication Costs	5.000,00	0,00	5.000,00	2,13
9 Dissemination	2.000,00	0,00	2.000,00	0,85
10 Overheads	16.450,00	0,00	16.450,00	7,00
11 Coordination Costs	not permitted	not permitted	not permitted	0,00
Total	315.000,00	80.000,00	235.000,00	100,00

Budget Justification
1 Staff Salary	Co-funding with man months of staff personnel salaries as detailed in the budget section
2 Researchers' Contracts	Fellowships for 2 new young U40 fellows to be hired for the 2 years duration of the project calculated at the cost of 25.000€/fellowship/year
3a.1 Equipment (Leasing - Rent)	N/A
3a.2 Equipment (buying)	N/A
3b Supplies	Cytoplex and RNA-seq analyses. kits for RNA and xxXXX extraction, probes for qRT-PCR. Antibodies and kits for flow cytometry and IHC.Plastics and reagents for storing / freezing and embedding paraffin tissues. Kits for amplification of PDX models.
3c Model Costs	Purchase of C57Bl/6, CD1 and NSG mice as detailed in the relevant project's sections, costs for mice transport, handling and feeding
4 Subcontracts	N/A
5 Patient Costs	N/A
6 IT Services and Data Bases	N/A
7 Travels	Calculated based in the participation of 2 members of the team to one national or international (european) congress with an average cost of 1000 € per congress per person
8 Publication Costs	Submission and publication costs of manuscripts on international peer-reviewed open access journals. Calculated as 2500€ average per publication, two publications
9 Dissemination	Registration fees for national or international meetings, and for printing posters, as the results obtained require dissemination among the scientific community and the general public. Calculated as 2000€ average
10 Overheads	Calculated as flat 7%
11 Coordination Costs	N/A

Principal Investigator Data

Cognome: XXXXXXXXX Nome: XXXXXXX

Genere: M

Codice fiscale: XXXXXX00X00X000X Documento: Carta d'identità, Numero: AV1756310 Data di nascita: 22/09/1954

Luogo di nascita: Napoli Provincia di nascita: NA

Indirizzo lavorativo: Xxx Xxxx Xxxxxxxx 00 Città: Roma

CAP: 00144

Provincia: RM

Email: xxxxxxx00.xxxxxxxxx@xxxxx.xxx Altra email: xxxxxxx.xxxxxxxxx@xxx.xx Telefono: x000000000000

Altro telefono: x00 00 0000 0000 Qualifica: Direttore Scientifico Struttura: Direzione Scientifica

Istituzione: IRCCS Istituto Nazionale Tumori Regina Elens Datore/ente di lavoro? Yes

Datore/ente di lavoro SSN? Yes Nome datore/ente di lavoro non SSN:

Nome istituzione SSN: IRCCS Istituto Nazionale Tumori Regina Xxxxx Tipo contratto: Lavoro Subordinato a Tempo determinato Full-TIME

Con l'invio della presente proposta si dichiara che la stessa o parti significative di essa non sono oggetto di altri finanziamenti pubblici o privati e che di conseguenza vi è assenza del c.d. doppio finanziamento ai sensi dell'art. 9 del Regolamento (UE) 2021/241, ossia che non ci sia una duplicazione del finanziamento degli stessi costi da parte di altri programmi dell'Unione, nonché con risorse ordinarie da Bilancio statale.

By submitting this proposal, I declare that no significant part or parts of it are recipient of any other public or private funding and that consequently there isn't any so-called double financing pursuant to art. 9 of Regulation (EU) 2021/241, i.e. that there is no duplication in the financing of the same costs by other Euopean Union programs or any other ordinary resources from the State budget.

Project validation result

Months

AIMS (A) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

A 1 Formulation/characterization of LNP-miRs

Formulations via microfluidic mixing

Purification of LNP-miRs via tangential flow filtration

A 2 Experiments in in vitro models

Experiments in ex vivo models

Experiments in in vivo models

A 3 PDX collection/characterization

Pharmacokinetic-pharmacodynamic studies

Drug efficacy studies in PDX and analyses of tumors

A375

M14

M14

A375

** *

1.0

M14

*

**

0.5

0.0

FC xxXXX Res vs Sens

FC xxXXX Res vs Sens

A

1.0

0.5

Sens

miR-199b-5p

miR-204-5p

Sens

miR-199b-5p

miR-204-5p

0.0

B

A375

*

VEGFA pg/mL

800

400

200

*

TGFβ1 pg/mL

100

300

***

CCL5 pg/mL

150

150

**

CXCL2 pg/mL

100

Res

Sens

50

Res

Sens

Res

M14

0

**

Sens

1000

0

*

TGFβ1 pg/mL

600

0

***

CCL5 pg/mL

800

0

***

CXCL2 pg/mL

Sens

Res

600

VEGFA pg/mL

500

300

400

300

Sens

Res

Sens

Res

Sens

Res

Sens

Res

0 0 0 0

Figure 1 A Quantification of miR-199b-5p and miR-204-5p by using qRT–PCR in A375 and M14 cells res vs. sens counterparts. B Elisa assays measuring VEGFA, TGFβ1, CCL5 and CXCL2 soluble levels in cell media deriving from A375 (upper panels) and M14 (lower panels) cells. *p < 0.05; **p < 0.01; and

***p < 0.001. qRT-PCR data are represented as the mean of three independent experiments ± SD; Xxxxx results are expressed as mean of at least three independent experiments ±SEM.

LNPs

A LNPs delivering miR-204-5p+miR-199b-5p

xxXXX/mRNA evaluation

MAPKi-res

B 100

A375

A375

*

*

FC xxXXX LNP-miR vs Scr

80

60

40

20

M14

800

M14

**

*

FC xxXXX LNP-miR vs Scr

600

400

miR-199b-5p

miR-204-5p

200

C

A375 res

melanoma cells

VEGFA pg/mL

800

400

LNP-Scr

0

48hrs

CM analysis Cell viability

*

*

TGFβ1 pg/mL

200

100

LNP-Scr

0

0

*

*

CCL5 pg/mL

Res

300

150

LNP-Scr

LNP-miR

LNP-miR

0

0

*

CXCL2 pg/mL

Res

miR-199b-5p

miR-204-5p

150

100

50

LNP-Scr

0

*

*

LNP-miR

LNP-miR

M14 res

**

1000

TGFβ1 pg/mL

600

*

CCL5 pg/mL

800

CXCL2 pg/mL

600

VEGFA pg/mL

500

300

400

300

LNP-Scr

LNP-Scr

LNP-miR

LNP-Scr

LNP-miR

LNP-Scr

LNP-miR

0 0 0 0

LNP-miR

D A375 res M14 res

Figure 2 A Schematic illustration of the biological assays carried out upon melanoma cells exposure to LNPs. B Quantification of miR-199b-5p and miR-204-5p by using qRT–PCR in res A375 and M14 cells following LNPs’ treatments. C Elisa assays measuring VEGFA, TGFβ1, CCL5 and CXCL2 soluble levels in cell media coming from A375 res (upper panels) and M14 res (lower panels) cells upon LNPs’ exposure. D Cell viability evaluation by measuring ATP content in A375 and M14 res cells untreated, treated with Dabrafenib (BRAFi) alone or in combination with the MEKi, i.e. Trametinib (MAPKi) in the presence of LNP-Scr or LNP-miR. *p < 0.05; **p < 0.01; and ***p

< 0.001; ****p < 0.0001 qRT-PCR data are represented as mean of three independent experiments ± SD; Xxxxx results are expressed as the mean of at least three independent experiments ±SEM.

1.5

   **

*

*

% of ATP content

1.0

0.5

0.0

         NS

       ***

  ****

****

***

% of ATP content

1.5

1.0

0.5

Unt

MAPKi+LNP-Scr

MAPKi+LNP-miR

BRAFi

Unt

MAPKi+LNP-Scr

MAPKi+LNP-miR

BRAFi

0.0

% of THP-1 migrated cells

A

**

250

200

150

100

50

CM sens

CM Res

0

A375 res

*

% of THP-1 migrated cells

250

200

150

100

50

CM sens

CM Res

LNP-miR

LNP-miR

0

M14 res B

% of THP-1 migrated cells

120

100

80

60

40

20

0

CM A375 res CM M14 res

**

**

% of THP-1 migrated cells

120

100

80

60

40

20

LNP-Scr

LNP-Scr

0

mRNA induction in THP-1 (%)

C

600

400

200

0

CD206

XX000

XXX0

XXX00

XX-00

CD86

-200

CM A375 res

**

*

Res vs Sens Res LNP-miR

*

*

*

*

*

**

*

**

**

***

vs Scr

200

mRNA induction in THP-1 (%)

100

0

CD206

XX000

XXX0

XXX00

XX-00

XX00

XX-00

XXX-0

XX-00

XXX-2

-100

CM M14 res

*

Res vs Sens Res LNP-miR

*

*

*

*

**

**

*

*

** **

vs Scr

M2 M1 M2 M1

Figure 3 A The migration of THP-1 monocytes in Transwell xxxxxxxx was induced after exposure to cell media coming from sens vs res A375 or M14 melanoma cells. B Cell media coming from resistant melanoma cells treated with LNP-Scr or LNP-miR and then used to induce the migration of THP-1 cells as reported in A. C qRT–PCR of the indicated M2 or M1 macrophages markers performed on THP-1 cells exposed to CM coming from res/sens melanoma cells (black bars) or from resistant cells treated with LNP-Scr or LNP-miR (green bars). *p < 0.05; **p < 0.01. qRT-PCR data are represented as the mean of at least three independent experiments ± SEM; migration results are expressed as the mean of three independent experiments ± SD.

Cell Randomization & Injection treatments

1 12 23 34 45 56 67 7

8 89 910110111121213131414

1511561167117811891920202121

2222232234224522562627272828

292390330131

A B

tumor mm3

LNPs

Mice sacrifice &

qRT-PCR

A375

Tumor measure, mice sacrifice & qRT-PCR

800

LNP-Scr LNP-miR

20 μg

40 μg

NS

*

600

400

200

0

96h

0

Log2 fold induction

C 10

miR-199b-5p

***

20 ug

40 ug

10 miR-204-5p

Log2 fold induction

* *** * *

20 ug

40 ug

NS

Figure 4. A Schematic illustration of the set up

5 NS

0

24h

-5

5 VEGFA

0

20 ug

40 ug

5

0

5

0

NS

NS

*

NS

*

*

NS

NS

Log2 fold induction

D

-

*

*

NS

NS *

48h

72h

96h

NS

5

0

24h

48h

-5

Log2 fold induction

5 BCL2 0

-5 NS NS

NS *

72h

96h

NS

NS

20 ug

40 ug

*

studies performed on A375 xenograft model. B Bar plots showing tumor volumes of mice injected with A375 cells (n=3) and treated with 20 mg or 40 mg of LNP-Scr o r L N P - m i R s . C Q u a n t i f i c a t i o n o f miR-199b-5p (left graph) and miR-204-5p (right graph) in mice treated as previously reported by using qRT–PCR at different time points (i.e. from 24h to 96 h). D Quantification of VEGFA (left graph) and

-1 -10

-1 -15

24h

48h

72h

96h

24h

48h

72h

96h

-2 -20

* **

*** **

BCL2 (right graph) in tumors by using qRT– PCR as reported in A. *p < 0.05; **p

< 0.01; and ***p < 0.001. Results are represented as the mean of three independent experiments ± SD.

A

LNPs

0

MAPKi

Cell Injection

Randomization

1 12 23 34 45 56 67 7

8 89 910110111121213131414

1511561167117811891920202121

2222232234224522562627272828

292390330131

measures

Mice sacrifice IHC analyses

C

B

Treatments &

A375

1200

tumor mm3

1000

800

600

400

200

0

A375

LNP-Scr LNP-miR

LNP-Scr+MAPKi LNP-miR+MAPKi

*

*

*

*

1

2

3

Weeks of treatment

1000

tumor mm3

800

600

400

200

0

0

M14

M14

LNP-Scr LNP-miR

LNP-Scr+MAPKi LNP-miR+MAPKi

*

*

1

2

3

Weeks of treatment

Log2 fold tumor volume

4 LNP-Scr+MAPKi

16% stable

66% growth

2

LNP-miR+MAPKi

3 LNP-Scr+MAPKi LNP-miR+MAPKi

Log2 fold tumor volume

10% stable

20% stable

2 40% growth

p=0,0489

0

28% growth 1

14% stable

0

p=0,0419

10% growth

16% regression

-2

57% regression

-1 50% regression

-2

70% regression

Week 3 of treatment

Figure 5 A Schematic workflow of the in vivo experiments performed with A375 or M14 melanoma xenografts treated with LNPs delivering miR-204-5p/miR-199b-5p and/or MAPKi. B Bar plots showing the measures of the tumor volumes relative to the mice injected with A375 (n=7) or M14 (n=10) cells and treated with LNP-Scr, LNP-miR or their combination with MAPKi. C The percentage of tumor shrinkage calculated respect to the initial volumes in mice injected with A375 (left panel) or M14 (right panel) belonging to the groups receiving LNP-Scr+MAPKi o r LNP-miR+MAPKi . D Immunohistochemistry (IHC) analyses of macrophage infiltration within the A375 and M14 derived tumors using the F4/80 antibody. *p

< 0.05. All in vivo results are represented as the mean (n=7 for A375; n=10 for M14) ± SEM.

Week 3 of treatment

D

% of F4/80 over tumor cells

% of F4/80 over tumor cells

X000 X00

XX

*

NS

*

30 2.5

2.0

20 1.5

10 1.0

0.5

LNP-Scr

LNP-miR

LNP-Scr+MAPKi

LNP-miR+MAPKi

LNP-Scr

LNP-miR

LNP-Scr+MAPKi

LNP-miR+MAPKi

0 0.0

F4/80 IHC (A375)

F4/80 IHC (M14)

20x

20x

LNP-Scr

20x

20x

LNP-miR

20x

20x

LNP-Scr+MAPKi

20x

20x

LNP-miR+MAPKi

Figure 6 Representative images of Immunohistochemistry (IHC) analyses by F4/80 antibody determining macrophage infiltration in tumor masses derived after injection of A375 (left panels) and M14 (right panels) cells in response to the indicated treatments. Original magnification = ×20.

35 LNP-Scr

LNP-miR

Mice weight

30

25

20

0

1

15

A375

LNP-Scr+MAPKi LNP-miR+MAPKi

30 LNP-Scr 25

Mice weight

20

0

1

15

M14

LNP-miR

LNP-miR+MAPKi

LNP-Scr+MAPKi

2

3

2

3

Weeks of treatment Weeks of treatment

Figure 7 Body weight measure of mice injected with A375 and M14 cells. A Mouse weight determination as a parameter to monitor animal suffering in response to the indicated treatments.

Formulation	Mean	PI ± SD	ZP	Actual	EE% ±
	(nm) ±		(mV ± SD)	loading	SD
	SD			(mg
				xxXXX/mg
				lipids)
LNP-Scr	137.3 ±	0.12 ± 0.02	-15.2 ± 3.4	186.4 ±	93.2 ±
	7.2			0.004	2.3
LNP-miR-204-5p/199b-5p	162.2 ±	0.16 ± 0.01	-27.2 ± 5.4	184.4 ±	92.2 ±
	1.6			0.002	3.2

Table 1. Characteristics of LNP-Scr or encapsulating miR-204-5p/199b-5p.