Animal Models. 8 2.6 Research Contributions........................................................9
Animal Models. Studies in the Research with respect to animal models in the Field shall be done in the U.S. and in Japan with the RMC's approval. Signal shall be responsible for the conduct of any such studies done in the U.S., and Nippon Kayaku shall be responsible for the conduct of any such studies done in Japan. Costs of animal model studies conducted in the U.S. and in Japan shall be shared equally by the Parties; provided, however, that, at Nippon Kayaku's option and expense, Nippon Kayaku may conduct any such animal model studies Signal does not agree to do.
Animal Models. Large animals such as dogs, pigs, and non-human primates, have been used in gene therapy 104 for several neuromuscular disorders such as myopathies, Xxxxxxx dystrophy, or Huntington 105–108, lysosome storage disorders 109,110, eye diseases 111,112, or cystic fibrosis 113. These large models have been used to assess efficacy, dosage, route of administration, and safety. Although large animal models for immunodeficiency have been described 114–116, immunodeficient mice are still the most used preclinical models to study gene therapy for immunodeficiencies like SCID. Due to the broad range of available immunocompromised mouse models, including the “humanized” mouse model, human patient cells can be xenografted and directly tested in vivo. Moreover, different genetic mouse models have been developed to mimic the different forms of SCID described in humans, such as ADA-SCID 19,117,118, X-linked (IL2rg)-SCID 9,119, Artemis-SCID 10–12 or RAG1/2-SCID 120,121. Importantly, these mice present a similar immunodeficient phenotype as found in humans, such as the Rag1-deficient mouse model, which present a full block at the early stages of T (DN2) and B (pre-B) cell development allowing close monitoring of the effects of the gene therapy in their development. Additionally, RAG1 and RAG2 hypomorphic SCID models are also available 122–125, allowing to study gene therapy in a wider range of immunodeficiencies with one same strategy. For example, analyzing whether the same vector can be used to correct both full RAG1-SCID and hypomorphic RAG1-SCID. Unfortunately, other SCID mouse models, such as X-linked (IL7r)-SCID does not reproduce the human setting as the mouse model has an extra B cell block that is not observed in humans 126. With the development of new editing tools (zinc-finger nucleases, TALENs, CRISPR-Cas9), transgenic mice can be generated to reproduce SCID phenotypes that do not have an established animal model yet. Even though we can find useful mouse models to study the efficacy of the developed gene therapy for immunodeficiencies, the gap between the mouse and the human physiological and pathological mechanisms is still substantial. The most recent achievement to overcome this gap is the development of “humanized mouse models”; immunodeficient mice such as nude or NSG mice carrying functioning human genes, human cells, or human tissues/organs. Importantly, these immunodeficient mice allow engraftment of functional human immune cells 127,128, enabling refined ...
Animal Models. Several different transgenic animal models were used in this study (Figure 1). En1cre/+and Foxp2Flp/+ animals were provided by Xxxxxx Xxxxxxxx and Xxxxxx Xxxxxxx, respectively. R26td+/td+ and R26tdTom/DC animals were obtained from Xxxxxxx Laboratories. En1cre/+:R26td+/td+ females were crossed with Foxp2Flp/+:R26DC/DC males to produce En1Cre/+: Foxp2Flp/+ : R26tdTom/DC animals. We observed aberrant tdtomato labeling of cells outside the ventral horn when En1cre/+:R26td+/td+ males were crossed with Foxp2Flp/+:R26DC/DC females, so we did not pair animals in this way. Thus, a cre-lox recombination system facilitated expression of the reporter gene tdTomato in all En1 expressing V1-INs. Cre expression from the En1 locus removes a floxed stop signal upstream of the tdtomato gene on the Rosa26 locus. Labeling of the intersectional population of Foxp2-expressing V1-INs with EGFP is conditional to the expression of both Cre and Flp recombinases (dual conditional, DC) from the En1 and Foxp2 loci, respectively. In DC reporter animals two STOP signals are present upstream of the EGFP gene on the Rosa26 locus: one flanked by FRT sites, and the other flanked by loxP sites. Cre and Flp-mediated recombination will remove these STOP signs and allow transcription of EGFP.
Animal Models. Animal models are useful in demonstrating the biological plausibility of certain hypotheses, especially when ethical limitations and potential psychological complications exist. A study by Xxxxx et al., which was first of its kind, examined the implications of cigarette smoke on the immune status of auto-immune-prone mice. The authors hypothesized that disease outcomes would be accelerated or exacerbated among animals that were predisposed to SLE if smoking did indeed have a fundamental causative relationship with the disease. Female MRL/Mp-lpr/lpr mice were assigned to groups that were exposed to different levels of cigarette smoke (0, 100 mg or 200 mg TPM/m3) over the course of 4 weeks. Biological samples, which contained protein, IgG, and IgM were collected were collected every 2-4 weeks for 4 months (Xxxxx, Xxxxxxxxx et al. 2005). The authors went on to complement their animal models with human data. Newly diagnosed SLE patients that had not begun treatment with immunosuppressive medications were used in the study sample (n=119). This study group consisted of 88 non-smokers and 31 current smokers. It was important to capture the SLE patients pre- treatment in order to rule out confounding by therapeutic interventions. SLE disease activity and severity were measured by the SLEDAI and the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SLICC/ACRDI) respectively. Questionnaires were used to determine smoking statuses (current, former, and never), and autoantibody determinations were performed (Xxxxx, Xxxxxxxxx et al. 2005). The results of the Xxxxx et al. study were surprising. Unlike the meta-analysis performed by Xxxxxxxxxxx et al., this study demonstrated that exposure to cigarette smoke actually suppressed IgG autoantibody development, and that autoantibody expression was augmented only after smoking stopped. Generally speaking, the results from this study showed that lupus-prone mice that were exposed to cigarette smoke in their early lives suppressed IgG autoantibody development, but not necessarily total immunoglobulin levels. Another interesting finding from this study was that the SLE patients who were currently smoking at the time of diagnosis were more likely to have neuropsychiatric problems and polyserositis than the other recently-diagnosed SLE participants, which is consistent with the findings of Ghaussy et al. (Ghaussy, Xxxxxxx et al. 2003; Xxxxx, Xxxxxxxxx et al. 2005). From their finding...
Animal Models. In the final stage of medicinal chemistry efforts, compounds with the most promising properties will be tested in at least one animal model of acute inflammatory disease in order to select Preclinical Lead Compounds. The specific animal model(s) chosen for this process and the number of compounds to be tested will be determined by the Management Committee. Acute Anti-inflammatory Models: - [CONFIDENTIAL TREATMENT REQUESTED] - [CONFIDENTIAL TREATMENT REQUESTED] - [CONFIDENTIAL TREATMENT REQUESTED] - [CONFIDENTIAL TREATMENT REQUESTED] - [CONFIDENTIAL TREATMENT REQUESTED] No later than [CONFIDENTIAL TREATMENT REQUESTED] after the initiation of the Plan, the Management Committee will choose the number and type of acute and/or chronic disease models for testing efficacy of compounds with the most promising properties and will discuss the experimental protocols for the chosen models. [CONFIDENTIAL TREATMENT REQUESTED].
