The HD iPSC Consortium: Repeat Length Dependent Phenotypes for Assay Development

描述(申请人提供)：亨廷顿病(HD)IPS联盟，由ARRA资助，为期两年，汇集干细胞和HD研究领域的领导小组，以确定新创建的IPS细胞系是否表现出HD相关(即，取决于Gag长度的)表型。该联盟旨在利用新技术，使用非整合方法重新编程，并在当前HD iPS系列中有希望的表型，为亨廷顿病(HD)的药物开发开发强有力和有效的分析方法，HD是一种致命的神经退行性疾病，目前没有治疗方法。HD患者的皮肤细胞可以重新编程为多能细胞，然后分化为特定的神经细胞和神经胶质细胞类型，从而可以研究遗传损伤对易感人类细胞类型的影响。我们密切合作，建立了从三个具有广泛GAG重复序列(33、60和180)的HD IPS整合株产生标准化神经干细胞培养物(例如EZ球体)的新方法，这些细胞已显示出许多有希望的表型。目前的提议将通过使用最新的非集成IPS技术额外生产15个品系来延长这些研究，并将应用新的分化和遗传标记协议来进一步优化系统。这笔资金将继续发挥这个非常专注的财团的显著协同作用，该财团(I)在创新的HD研究和合作方面有着良好的记录，(Ii)准备继续利用HD患者的诱导多潜能干细胞(IPS)进行尖端研究，并致力于广泛传播研究结果、方案和IPS系列，以及(Iii)与GOL合作的一组研究人员，以优化神经元特异性分化方案。持续的资金将加快对IPS系列的协调分析，并利用互补的、协同的技能集，比任何组织单独使用都能更快地推动该领域的发展。我们的最终目标是开发和验证使用&gt；96 Well格式的适合高含量/高吞吐量筛选方法的Gag重复长度依赖表型的方法和分析。拟议的研究将评估iPS细胞技术在建立HD模型和药物发现方面的能力。HD的单基因性质和具有一定范围的Gag重复长度的等位基因序列的存在赋予了建立神经退行性疾病模型的巨大力量。这些细胞系将成为学术团体和制药公司研究发病机制和测试HD实验疗法的重要资源。

DESCRIPTION (provided by applicant): The Huntington's disease (HD) IPS consortium, funded with ARRA support for two years, brings together leading groups in stem cell and HD research to establish whether newly created IPS cell lines show HD-related (i.e., GAG length-dependent) phenotypes. This consortium aims to capitalize on new technologies to use non-integrating approaches for reprogramming and promising phenotypes in current HD iPS lines to develop robust and validated assays for drug development for Huntington's disease (HD), a fatal neurodegenerative condition with no current treatment. Skin cells from patients with HD can be reprogrammed to pluripotency and then differentiated into specific neuronal and glial cell types, permitting investigation of the effects of the genetic lesion in the susceptible human cell types. We have worked closely together and established novel methods of generating standardized neural stem cell cultures (e.g., EZ spheres) from three HD IPS integrating lines with a wide range of GAG repeats (33, 60 and 180), which have shown a number of promising phenotypes. The current proposal will extend these studies by producing 15 additional lines using the latest non-integrating IPS technology and will apply novel differentiation and genetic tagging protocols to further optimize the system. This funding would continue the significant synergy of this very focused consortium that (i) has a strong track record of innovative HD research and of working together, (ii) is poised to continue in cutting-edge research with induced pluripotent stem (IPS) cells derived from HD patients and is committed to broad distribution of findings, protocols and IPS lines, and (iii) is partnered with a group of investigators with the gol to optimize neuron specific differentiation protocols. Continued funding will accelerate the coordinated analysis of IPS lines and leverage the complementary, synergistic skill sets that will move the field forward more rapidly than would be possible by any group alone. Our ultimate goal is to develop and validate methods and assays using >96 well format for GAG repeat length-dependent phenotypes that are amenable to high content/throughput screening methods. The proposed studies will provide an assessment of the power of iPS cell technology for modeling HD, and for drug discovery. The monogenic nature of HD and the existence of allelic series of IPSCs with a range of GAG repeat lengths confer tremendous power to model neurodegenerative disease. These cell lines will be an essential resource for academic groups and pharmaceutical companies for studying pathogenesis and for testing experimental therapeutics for HD.