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

描述（由申请人提供）：亨廷顿氏病（HD）IPS联盟在ARRA的支持下资助了两年，汇集了干细胞和HD研究的领先小组，以确定新创建的IPS细胞系是否显示HD相关（即，GAG长度依赖性）表型。该联盟旨在利用新技术，在当前HD iPS系中使用非整合方法进行重编程和有希望的表型，以开发用于亨廷顿病（HD）药物开发的稳健且经验证的检测方法，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被重编程为多能性，然后分化为特定的神经元和神经胶质细胞类型，从而可以研究易感人类细胞类型中遗传病变的影响。我们密切合作，建立了产生标准化神经干细胞培养物的新方法（例如，EZ球）从三个HD IPS整合线与广泛的GAG重复序列（33，60和180），这已经显示了许多有前途的表型。目前的提案将通过使用最新的非整合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方案来进一步优化系统。这笔资金将继续这个非常集中的财团的显着协同作用，（i）具有创新HD研究和合作的良好记录，（ii）准备继续进行来自HD患者的诱导多能干细胞（IPS）的尖端研究，并致力于广泛传播研究结果，方案和IPS系，和（iii）与GOL的一组研究人员合作，以优化神经元特异性分化方案。 继续提供资金将加快对IPS系列的协调分析，并利用互补、协同的技能组合，使该领域比任何单独的小组都能更快地向前推进。 我们的最终目标是开发和验证使用>96孔格式的GAG重复序列长度依赖性表型的方法和测定法，这些方法和测定法适用于高含量/通量筛选方法。拟议的研究将评估iPS细胞技术用于HD建模和药物发现的能力。HD的单基因性质和具有一系列GAG重复长度的IPSC的等位基因系列的存在赋予了对神经退行性疾病建模的巨大能力。这些细胞系将成为学术团体和制药公司研究HD发病机制和测试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.