Novel Therapeutics for FSHD - Resources Core - Core C

项目总结 Resources Core将提供细胞、动物模型和生物信息学试剂、资源和数据库 支持韦尔斯通研究项目以及更大的FSHD和肌肉营养不良研究 社区。资源核心将继续建立一个主要的活体组织和细胞生物材料储存库 来自FSHD患者及其一级家庭成员，重点是无症状的FSHD家庭 研究对象(目标1)。患者来源的低传代CD56肌肉细胞和重新编程的IPSC肌源性细胞系 将成为验证项目1和DUX4中基因组临床研究中确定的修饰基因的资源 项目2和3中的途径研究以及药物发现和验证研究：来自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 以临床和基因验证的细胞系进行的统计支持的研究。资源核心将成为 为我们新开发的Wellstone FSHD动物模型提供新的资源，以支持动物药物研究，并将 动物和细胞FSHD肌肉病理的分子、组织病理学和生理学分析资源 模特们。Wellstone FSHD动物模型包括：1)DUX4注射和DUX4诱导斑马鱼模型 实现快速药物和吗啡测试，以调查发育和组织特异性机制 DUX4毒性(目标2)；2)首次获得三苯氧胺诱导的条件性DUX4转基因小鼠 严密控制的DUX4可诱导FSHD小鼠模型，表达可遗传的DUX4转基因并显示 诱导性肌病和病理生理学，非常适合测试候选FSHD药物(Aim3)；DUX4 斑马鱼和老鼠模型将被用来研究项目1、2和3中确定的FSHD修饰基因， 并在项目2和项目3中测试翻译后、低氧和HA信号通路抑制物。 异种移植提供了一种独特的FSHD动物模型，使研究阻断DUX4的药物和ASO成为可能 高度人源化、血管化和神经化的FSHD肌肉的功能，标准细胞无法实现 异种移植(目标4)。FSHD异种肌肉模型将用于优化DUX4的ASO基因敲除 以及在项目2和3中测试4-MU和候选HA和缺氧信号药物。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，规划良好- 为所有项目设计的、统计支持的研究，并支持GWAS和DNA甲基化分析， 包括与项目1的现有Wellstone数据的比较以及ASO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韦尔斯通中心的网站和 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.