Core 1: Muscular Dystrophy Cell and Serum Banking Core

肌营养不良细胞系和血清库核心 摘要 这个P50 CORT资助申请的主题是加速新基因的翻译。 从实验室到诊所。研究致病性变体或其 在各种肌营养不良症中的校正是使用 患者来源的肌细胞。来自肌营养不良症患者的肌细胞的增殖能力是 有限，并且获得细胞的过程（肌肉活检）是侵入性的。一种使用慢病毒载体用于 hTERT和MyoD都递送到成纤维细胞以产生肌原性成纤维细胞（下文称为 FibroMyoD）将克服这一瓶颈。肌营养不良症细胞系的总体目的和 血清库核心（MD-CLSB核心）将制备和储存人原代和永生化细胞 将支持每个CORT项目的生产线，以及用于探索性研究的血清和血浆库 和合作项目。总体目标将分为三个具体目标。目标1将创建一个 来自肌营养不良受试者的独特细胞系资源，通过皮肤成纤维细胞库获得， 活组织检查和创建FibroMyoD细胞系。目标2将建立血清生物库资源， 从临床患者和研究受试者中获得的样本，并将这些样本提供给CORT， 外部附属研究人员进行假设驱动和发现研究。最后，在目标3中，我们将开发 一种改进的转分化方案，从FibroMyoD系产生成熟肌纤维， 与成熟肌纤维相似该MD-CLSB核心将利用全国现有的专业知识， 儿童医院（NCH）研究所细胞系核心，由金L。McBride，MD.这 近10年来，由机构支持的共享资源一直在创建和储存细胞系， 在过去的三年里，已经为拟议的CORT主任的实验室建立了原代成纤维细胞系， 博士凯文·弗拉尼根。目前的建议将涉及对现有NCH细胞系的需求大幅增加 核心，以及CORT核心的建立将提供所需的额外资源， NCH和更广泛的肌肉研究社区。产生的细胞系和血清样品 存储将是一个宝贵的资源，为拟议的CORT的项目，和更广泛的肌肉 营养不良研究社区，直接与P50 CORT使命。

Muscular Dystrophy Cell Line and Serum Banking Core ABSTRACT The over-arching theme for this P50 CORT grant application is to accelerate the translation of novel genetic therapies from the bench into the clinic. A significant bottleneck in studying pathogenic variants or their correction in the various muscular dystrophies is the limiting factor of a muscle cell model system using patient derived myocytes. The proliferative capacity of muscle cells from muscular dystrophy patients is limited, and the process to obtain cells (muscle biopsy) is invasive. A technique using lentiviral vectors for both hTERT and MyoD delivery to the fibroblasts to create myogenic fibroblasts (hereafter called FibroMyoD) will overcome this bottleneck. The overall objective of the Muscular Dystrophy Cell Line and Serum Banking Core (MD-CLSB Core) will be preparing and banking human primary and immortalized cell lines that will support each of the CORT Projects, as well as for serum and plasma banking for exploratory and collaborative projects. The overall objective will be addressed in three specific aims. Aim 1 will create a unique cell line resource from muscular dystrophy subjects by banking dermal fibroblasts, obtained by skin biopsy, and creating FibroMyoD cell lines. Aim 2 will establish a serum biobank resource, through banking of samples obtained from clinic patients and research subjects, and make these samples available to CORT and external affiliated investigators for hypothesis-driven and discovery research. Finally, in Aim 3, we will develop an improved transdifferentiation protocol to generate mature myofibers from the FibroMyoD lines that more closely mimic mature myofibers. This MD-CLSB Core will leverage the existing expertise of the Nationwide Children's Hospital (NCH) Research Institute Cell Line Core, directed by Kim L. McBride, MD. This institutionally-supported shared resource has been creating and banking cell lines for nearly 10 years, and for the past three years has created primary fibroblast cell lines for the laboratory of the proposed CORT director, Dr. Kevin Flanigan. The current proposal will involve a greatly increased demand on the existing NCH Cell Line Core, and establishment of the CORT Core will provide the additional resources needed to serve as a resource to both the NCH and the broader muscle research community. The cell lines generated and serum samples stored will be a valuable resource for the projects of the proposed CORT, and the broader muscular dystrophy research community, in direct alignment with the P50 CORT mission.