Transgenic marmosets for translational stem cell research

用于转化干细胞研究的转基因狨猴

• 批准号: 9215707
• 负责人: MARINA EMBORG
• 金额: $ 67.04万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9215707
• 关键词: Advanced Development; Age; Aging; Alleles; Animal Disease Models; Animal Experiments; Animal Model; Animals; Area; Arthritis; Assisted Reproductive Technology; Behavioral; Biochemical; Biological Assay; Biomedical Research; Birth; Callithrix; Callithrix jacchus jacchus; Categories; Cell Line; Cells; Clinical Trials; Collaborations; Consultations; Development; Diabetes Mellitus; Disease; Disease model; Embryo; Embryology; Engraftment; Experimental Models; Exposure to; Fibroblasts; Fostering; Functional disorder; Future; Gene Transfer Techniques; Generations; Genes; Goals; Green Fluorescent Proteins; Health; Home environment; Housing; Human; Huntington Disease; Huntington gene; Image; Immunology; In Vitro; Institutes; Institution; LRRK2 gene; Lentivirus Vector; Longevity; Macaca; Macaca mulatta; Medical; Mission; Modeling; Monkeys; Monoclonal Antibody R24; Mutation; National Institute of Child Health and Human Development; National Institute of Neurological Disorders and Stroke; Neurodegenerative Disorders; Neurologic; North America; Operative Surgical Procedures; Outcome; Parkinson Disease; Patients; Physics; Pluripotent Stem Cells; Preclinical Testing; Primates; Procedures; Production; Regenerative Medicine; Reporting; Reproductive Medicine; Research; Research Personnel; Research Priority; Resources; Safety; Scientist; Services; Source; Stem Cell Research; Stem cells; Technology; Testing; Therapeutic; Toxin; Transgenes; Transgenic Organisms; Translational Research; Transplantation; United States National Institutes of Health; Variant; Wisconsin; Work; age related; base; cohort; efficacy evaluation; enhanced green fluorescent protein; genome editing; human disease; imaging study; induced pluripotent stem cell; interest; nervous system disorder; nonhuman primate; novel strategies; offspring; overexpression; personalized medicine; pre-clinical; public health relevance; relating to nervous system; reproductive; stem cell therapy; therapeutic evaluation; tool; transgene expression; translational approach; transmission process

DESCRIPTION (provided by applicant): Nonhuman primates (NHP) offer the most appropriate experimental model for many areas of human biomedical research, particularly in reproductive medicine, immunology and transplantation, and neurological disease. Furthermore, monkey induced pluripotent stem cells (iPSCs) are envisioned as a valuable renewable resource of cells for preclinical testing of personalized regenerative medicine approaches. Based on initial advances in common marmoset assisted reproductive technologies (ARTs) developed at the Wisconsin National Primate Research Center (WNPRC), the production of transgenic common marmosets expressing green fluorescent protein, including the demonstration of germline transmission, has been reported. Marmoset monkeys present several advantages for transgenic approaches, including the ability to routinely carry multiple offspring (rapidly increasing cohort size), facile reproductive management, and a shorter lifespan, which facilitates the study of age-related diseases such as diabetes, arthritis and Parkinson's disease (PD). In regards to PD, identification of specific alleles of the leucine rich repeat kinase 2 (LRRK2) in familial and sporadic cases of PD supports the development of a NHP model expressing these variants, as proof-of-principle for the contribution of human alleles to disease pathophysiology. We propose to advance the development of animal disease models for stem cell-based therapeutic applications with two Specific Aims: Specific Aim 1. To optimize reprogramming of iPSCs from common marmoset fibroblasts, to use transgene expression and genomic editing to define the effect of alleles associated with PD on in vitro neural differentiation in isogenic iPSC lines, and to use genomic editing tools to define the feasibility, efficiency and accuracy of genomic editing in IVF-derived common marmoset embryos. Specific Aim 2. To produce transgenic marmoset monkeys carrying wild-type and PD-associated human LRRK2 alleles and evaluate a "proof-of-principle" cohort for PD pathophysiology. The long-term goal of this work is to provide investigators with marmoset iPSC lines, and genetically modified common marmosets as platforms for translational research, particularly in the treatment of diseases for which primate species are the most suitable models. Animal experiments will be performed at WNPRC, which is one of the few facilities in North America housing an experimental common marmoset colony, and the only Center where marmosets, primate embryology, and cutting edge neurological translational models are actively being used to test therapies for human disease. The proposed generation and analysis of transgenic monkeys and associated modified iPSCs will provide a platform to create disease specific models and cells, and develop tests of therapeutic approaches, including personalized medicine using engraftment of iPSCs.

描述（由申请人提供）：非人类灵长类动物（NHP）为人类生物医学研究的许多领域提供了最合适的实验模型，特别是在生殖医学、免疫学和移植以及神经系统疾病方面。此外，猴诱导多能干细胞（iPSC）被认为是一种宝贵的可再生细胞资源，可用于个性化再生医学方法的临床前测试。基于威斯康星国家灵长类研究中心 (WNPRC) 开发的普通狨猴辅助生殖技术 (ART) 的初步进展，已报道了表达绿色荧光蛋白的转基因普通狨猴的生产，包括生殖系传播的演示。狨猴具有转基因方法的几个优点，包括能够定期携带多个后代（群体规模迅速增加）、方便的生殖管理和较短的寿命，这有助于研究糖尿病、关节炎和帕金森病（PD）等与年龄相关的疾病。关于帕金森病，在家族性和散发性帕金森病病例中鉴定出富含亮氨酸重复激酶 2 (LRRK2) 的特定等位基因，支持开发表达这些变异的 NHP 模型，作为人类等位基因对疾病病理生理学贡献的原理证明。我们建议通过两个具体目标推进基于干细胞的治疗应用的动物疾病模型的开发： 具体目标 1. 优化普通狨猴成纤维细胞的 iPSC 重编程，使用转基因表达和基因组编辑来定义与 PD 相关的等位基因对同基因 iPSC 系体外神经分化的影响，以及 使用基因组编辑工具来确定体外受精衍生的普通狨猴胚胎基因组编辑的可行性、效率和准确性。具体目标 2. 产生携带野生型和 PD 相关人类 LRRK2 等位基因的转基因狨猴，并评估 PD 病理生理学的“原理验证”队列。这项工作的长期目标是为研究人员提供狨猴 iPSC 系和转基因普通狨猴作为转化研究的平台，特别是在治疗以灵长类动物为最合适模型的疾病方面。动物实验将在 WNPRC 进行，该中心是北美为数不多的拥有普通狨猴实验群体的设施之一，也是唯一一个积极利用狨猴、灵长类胚胎学和尖端神经转化模型来测试人类疾病疗法的中心。拟议的转基因猴和相关修饰 iPSC 的生成和分析将提供一个平台来创建疾病特异性模型和细胞，并开​​发治疗方法测试，包括使用 iPSC 移植的个性化医疗。