Core 1: Muscular Dystrophy Cell and Serum Banking Core

肌营养不良细胞系与血清银行核心 摘要 这个P50 Cort奖助金申请的总体主题是加速新基因的翻译 从替补席到诊所的治疗方法。研究致病变种或其变种的一个重大瓶颈 在各种肌肉营养不良中的矫正是使用 患者来源的肌细胞。肌营养不良症患者肌细胞的增殖能力 而且获取细胞的过程(肌肉活组织检查)是侵入性的。一种使用慢病毒载体的技术 HTERT和MyoD都可以传递给成纤维细胞以产生肌源性成纤维细胞(以下称为 FibroMyoD)将克服这一瓶颈。肌营养不良细胞系的总体目标和 血清银行核心(MD-CLSB核心)将准备和银行人类原代和永生化细胞 将支持每个CORT项目的线路，以及用于探索性的血清和血浆银行 和协作项目。总体目标将通过三个具体目标来实现。目标1将创建一个 肌营养不良患者皮肤来源独特的真皮成纤维细胞来源 活组织检查，并建立纤维肌肉细胞系。目标2将建立一个血清生物库资源，通过银行 从临床患者和研究对象获得的样本，并将这些样本提供给CORT和 外部附属调查人员，用于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系产生成熟的肌纤维，比 近乎模仿成熟的肌纤维。该MD-CLSB核心将利用全国范围内的现有专业知识 儿童医院(NCH)研究所细胞系核心，导演：Kim L.McBride，MD。这 制度支持的共享资源已经创建和存储细胞系近10年，并且 在过去的三年里，为拟任CORT主任的实验室培养了原代成纤维细胞系， 凯文·弗拉尼根医生。目前的建议将涉及对现有NCH细胞系的需求大幅增加 核心，CORT核心的建立将提供作为资源所需的额外资源 NCH和更广泛的肌肉研究社区。所产生的细胞系和血清样本 STORAGE将成为拟议的CORT项目的宝贵资源，以及更广泛的肌肉 营养不良研究社区，与P50 CORT任务直接一致。

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.