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

描述（由申请人提供）：亨廷顿病（HD） IPS联盟由ARRA资助，为期两年，汇集了干细胞和HD研究的领先团队，以确定新创建的IPS细胞系是否显示HD相关（即GAG长度依赖）表型。该联盟旨在利用新技术，利用非整合方法对当前亨廷顿舞蹈病（HD）的iPS系进行重编程和有希望的表型，以开发强大且有效的亨廷顿舞蹈病（HD）药物开发分析。亨廷顿舞蹈病是一种致命的神经退行性疾病，目前尚无治疗方法。来自HD患者的皮肤细胞可以被重新编程为多能性，然后分化为特定的神经元和神经胶质细胞类型，从而允许研究遗传病变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整合系中生成标准化神经干细胞培养物（例如EZ球）的新方法，这些细胞已显示出许多有希望的表型。目前的提案将通过使用最新的非集成IPS技术生产15个额外的品系来扩展这些研究，并将应用新的分化和遗传标记协议来进一步优化系统。这笔资金将继续这个非常专注的联盟的重要协同作用，(i)在创新的HD研究和合作方面有着良好的记录，（ii）准备继续进行来自HD患者的诱导多能干细胞（IPS）的前沿研究，并致力于广泛分发研究结果、方案和IPS细胞系，以及（iii）与一组研究人员合作，目标是优化神经元特异性分化方案。持续的资金将加速对IPS品系的协调分析，并利用互补、协同的技能组合，使该领域比任何单独的团体都能更快地向前发展。我们的最终目标是开发和验证使用bbb96孔格式的GAG重复长度依赖表型的方法和分析，这些方法和分析适用于高含量/高通量筛选方法。拟议的研究将提供对iPS细胞技术在HD建模和药物发现方面的能力的评估。HD的单基因性质和具有GAG重复长度范围的IPSCs等位基因系列的存在赋予了巨大的力量来模拟神经退行性疾病。这些细胞系将成为学术团体和制药公司研究HD发病机制和试验治疗方法的重要资源。

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. PUBLIC HEALTH RELEVANCE: Huntington's disease is a significant neurodegenerative disease, whose relatively simple and unique known genetic cause, a GAG expansion in the HD gene correlated with severity and onset of clinical symptoms, makes it particularly suited to therapeutic development. Preliminary studies indicate that GAG length dependent phenotypes occur at all stages of differentiation, from IPSC through to mature neurons, providing the feasibility of robust assay development for drug discovery. These assays will allow academic groups and pharmaceutical companies to study pathogenesis and test experimental therapeutics for HD, which will significantly advance both our understanding of HD and potential treatments for this devastating and currently untreatable disease.