Novel Therapeutics for FSHD - Resources Core - Core C

项目摘要 资源中心将提供细胞、动物模型和生物信息学试剂、资源和数据库， 支持Wellstone研究项目以及更大的FSHD和肌肉萎缩症研究 社区.资源核心将继续建立一个主要的生物材料库的活检和细胞 从FSHD患者及其一级家庭成员，重点是FSHD家庭， 主题（目标1）。患者来源的低传代CD 56+肌细胞和重编程iPSC肌细胞系 将成为验证项目1和DUX 4中基因组临床研究中鉴定的修饰基因的资源 途径研究以及项目2和项目3中的药物发现和验证研究。FSHD的肌源性细胞 家庭队列将继续分发给FSHD研究人员和行业合作者，以支持 临床和遗传学验证的细胞系的统计学功效研究。核心资源将成为 我们新开发的Wellstone FSHD动物模型的新资源，使动物药物研究， 一个资源的分子，组织病理学和生理学分析FSHD肌肉病理在动物和细胞 模型Wellstone FSHD动物模型包括：1）DUX 4注射和DUX 4诱导的斑马鱼模型 使快速药物和吗啉测试，以调查发展和组织特异性机制， DUX 4毒性（目标2）; 2）他莫昔芬诱导的条件DUX 4转基因小鼠，这是第一个同类 表达可遗传的DUX 4转基因并表现出 诱导性肌病和病理生理学，对于测试候选FSHD药物是理想的（Aim 3）; 3）. Dux4 斑马鱼和小鼠模型将用于研究项目1、2和3中鉴定的FSHD修饰基因， 并在项目2和3中测试翻译后、缺氧和HA信号通路抑制剂。FSHD肌 异种移植物提供了独特的FSHD动物模型，使得能够研究阻断DUX 4的药物和ASO 在高度人源化、血管化和神经支配的FSHD肌肉中发挥功能，这是标准细胞无法实现的 异种移植物（Aim 4）。FSHD异种移植肌肉模型将用于优化DUX 4的阿索敲低。 以及在项目2和3中测试4-MU和候选HA和缺氧信号传导药物。生物信息学和 统计核心将继续负责管理综合临床和实验数据库， 数据，对所有项目产生的数据进行生物信息学和统计分析， 为所有项目设计的统计学研究，并支持GWAS和DNA甲基化分析， 包括与项目1的现有Wellstone数据的比较和阿索研究的RNA-Seq数据分析， 项目3（目标5）。核心资源的可用性将通过Wellstone中心网站发布， 通过在FSHD患者和研究会议上的演讲。

PROJECT SUMMARY The Resources Core will provide cell, animal model and bioinformatic reagents, resources, and databases to support the Wellstone research projects as well as the greater FSHD and muscular dystrophy research communities. The Resources Core will continue to build a major biomaterials repository of biopsies and cells from FSHD patients and their first degree family members, with a focus on FSHD families with non-manifesting subjects (Aim 1). Patient-derived, low passage CD56+ muscle cells and reprogrammed iPSC myogenic lines will be a resource for validation of modifier genes identified in genomic clinical studies in Project 1 and in DUX4 pathway studies and in drug discovery and validation studies in Projects 2 and 3. Myogenic cells from FSHD family cohorts will continue to be distributed to FSHD researchers and industry collaborators to support statistically powered studies with clinically and genetically validated cell lines. The Resources Core will become a new resource for our newly developed Wellstone FSHD animal models to enable animal drug studies, and be a resource for molecular, histopathology and physiology assays of FSHD muscle pathology in animal and cell models. Wellstone FSHD animal models include: 1) DUX4 injection and DUX4-inducible zebrafish models enable rapid drug and morpholino testing to investigate developmental and tissue-specific mechanisms of DUX4 toxicity (Aim 2); 2) a tamoxifen-inducible conditional DUX4 transgenic mouse that is a first-of-its-kind tightly controlled DUX4 inducible FSHD mouse model that expresses a heritable DUX4 transgene and displays inducible myopathy and pathophysiology, ideal for testing of candidate FSHD drugs (Aim3); 3). DUX4 zebrafish and mouse models will be used to investigate FSHD modifier genes identified in Projects 1, 2 and 3, and to test posttranslational, hypoxia and HA signaling pathway inhibitors in Projects 2 and 3. FSHD muscle xenografts provide a unique FSHD animal model enabling investigations of drugs and ASOs that block DUX4 function in a highly humanized, vascularized and innervated FSHD muscle, not achieved with standard cell xenografts (Aim 4). The FSHD xenograft muscle model will be used for optimizing ASO knockdown of DUX4 and testing 4-MU and candidate HA and hypoxia signaling drugs in Projects 2 and 3. A bioinformatics and statistical core will continue to be responsible for managing databases of integrated clinical and experimental data, performing bioinformatic and statistical analyses of data generated in all Projects, planning well- designed, statistically powered studies for all Projects, and supporting GWAS and DNA methylation analyses, including comparison to existing Wellstone data for Project 1 and RNA-Seq data analysis of ASO studies in Project 3 (Aim 5). Availability of Core resources will be disseminated via the Wellstone Center website and through presentations at FSHD patient and research meetings.