Reprogramming and Directed Differentiation of Skeletal Muscle Cells from hPSCs.

hPSC 中骨骼肌细胞的重编程和定向分化。

• 批准号: 9335654
• 负责人: April D Pyle
• 金额: $ 32.07万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-20 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9335654
• 关键词: Aging; Antisense Oligonucleotides; Biology; Birth; Cachexia; Cell Compartmentation; Cell Differentiation process; Cell Line; Cell Therapy; Cell Transplantation; Cells; Cellular biology; Childhood; Clinical Trials; Cues; Cyclic GMP; Development; Disease; Duchenne muscular dystrophy; Dystrophin; Embryo; Embryonic Development; Engraftment; Evaluation; Foundations; Future; Genetic; Genome; Germ Layers; Goals; Grant; Growth; Growth Factor; Human; Human body; Immune; In Vitro; Incidence; Instruction; Knowledge; Laboratories; Lead; Lentivirus Vector; Light; Location; Maintenance; Mediating; Mesoderm; Modeling; Mus; Muscle; Muscle Cells; Muscle Development; Muscle Weakness; Muscular Atrophy; Muscular Dystrophies; Mutation; Myopathy; Nanostructures; PAX7 gene; Paraxial Mesoderm; Pathway interactions; Patients; Pattern; Population; Progenitor Cell Engraftment; Reading Frames; Recruitment Activity; Regenerative Medicine; Research; Role; SHH gene; Safety; Sampling; Signal Transduction; Skeletal Muscle; Somatic Cell; Source; Specific qualifier value; Stem cells; System; Testing; Therapeutic; Therapeutic Intervention; To specify; Transplantation; Tumorigenicity; Viral; Virus Diseases; Work; cell type; design; effective therapy; exon skipping; experimental study; genetic manipulation; human embryonic stem cell; human pluripotent stem cell; improved; in vivo; induced pluripotent stem cell; male; mini-dystrophin; molecular recognition; mouse model; myogenesis; nanoparticle; nanostructured; novel; novel strategies; overexpression; pre-clinical; progenitor; public health relevance; regenerative; regenerative therapy; repaired; research clinical testing; satellite cell; screening; self assembly; skeletal; skeletal muscle differentiation; small molecule; tool; transcription factor; tumorigenic

DESCRIPTION (provided by applicant): Human pluripotent stem cells (hPSCs) have enormous promise for regenerative medicine as they can differentiate into cells from all three embryonic germ layers. Further hPSCs provide a unique pre-clinical screening tool for evaluating disease mechanisms and therapies in patient samples. Despite the enormous potential of hPSC, many obstacles need to be overcome before the use of stem cells in cell-based therapy will be realized, including an incomplete knowledge of the growth factor cues regulating human skeletal muscle progenitor cell (SMPC) specification, growth and engraftment. In this proposal we will investigate the cellular cues required to direct skeletal muscle fate from hPSCs as seen during embryonic development. We will also develop the first screening approach to identify new regulators of human SMPC fate. It is imperative that we understand how to obtain SMPCs without genetic or viral manipulation. As an alternative approach we have developed a unique non-viral reprogramming platform, which results in superior delivery and reprogramming efficiency, and will be utilized to specify SMPCs without viral or genetic manipulation. We will compare the in vivo engraftment potential of growth factor directed versus reprogrammed SMPCs in a mouse model of DMD. The development of reprogramming strategies to direct SMPC fate and the identification of signals regulating muscle progenitor cell specification and maintenance could improve our basic understanding of human skeletal myogenesis as well as enhance our ability to generate scalable progenitors from humans for muscle disorders including DMD.

描述（由申请人提供）：人类多能干细胞（hPSC）在再生医学方面具有巨大的前景，因为它们可以分化为来自所有三个胚胎胚层的细胞。此外，hPSC提供了独特的临床前筛选工具，用于评估患者样品中的疾病机制和疗法。尽管hPSC具有巨大的潜力，但在实现干细胞在基于细胞的治疗中的应用之前，需要克服许多障碍，包括对调节人骨骼肌祖细胞（SMPC）特化、生长和植入的生长因子线索的不完全了解。在这个建议中，我们将研究细胞线索所需的直接骨骼肌的命运， 在胚胎发育期间观察到的hPSC。我们还将开发第一种筛选方法，以确定人类SMPC命运的新调控因子。我们必须了解如何在没有基因或病毒操纵的情况下获得SMPC。作为一种替代方法，我们开发了一种独特的非病毒重编程平台，该平台具有上级递送和重编程效率，并将用于指定SMPC，而无需病毒或遗传操作。我们将在DMD小鼠模型中比较生长因子定向与重编程SMPC的体内植入潜力。重编程策略的发展，以指导SMPC的命运和识别信号调节肌肉祖细胞的规范和维护，可以提高我们对人类骨骼肌发生的基本理解，以及提高我们的能力，从人类产生可扩展的祖细胞的肌肉疾病，包括DMD。