Interrogating functional and molecular properties of PAX7+ putative skeletal muscle stem/progenitor cells derived from human iPSCs of healthy donors and Duchenne muscular dystrophy patients

探究源自健康捐献者和杜氏肌营养不良症患者 iPSC 的 PAX7 推定骨骼肌干/祖细胞的功能和分子特性

• 批准号: 10161732
• 负责人: Gabsang Lee
• 金额: $ 34.85万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10161732
• 关键词: Address; Adopted; Adult; Aging; Autologous; CRISPR/Cas technology; Cachexia; Cell Line; Cell Maintenance; Cell Transplantation; Cells; Child; Clinical; Clinical Trials; Conceptions; DNA Sequence Alteration; Development; Disease; Disease model; Duchenne muscular dystrophy; Dystrophin; Ectopic Expression; Embryo; Environment; Event; Extracellular Matrix; Genes; Genetic; Genetic Diseases; Genetic Transcription; Healthcare Systems; Homologous Transplantation; Human; In Vitro; Injury; Knowledge; Length; Life; Membrane; Mesoderm; Methodology; Molecular; Mus; Muscle; Muscle Fibers; Muscle function; Muscle satellite cell; Muscular Atrophy; Muscular Dystrophies; Mutation; Myoblasts; Myopathy; Neuromuscular Diseases; Non-Insulin-Dependent Diabetes Mellitus; PAX7 gene; Patients; Phenotype; Phospholipids; Pluripotent Stem Cells; Population; Process; Proliferating; Property; Proteins; Regenerative Medicine; Regenerative capacity; Regulation; Reporter; Reporting; Research; Role; Site; Skeletal Muscle; Structure; System; Techniques; Teenagers; Telomere Shortening; Time; Transplantation; Traumatic injury; Untranslated RNA; Variant; Wasting Syndrome; base; blastomere structure; cancer cachexia; cell behavior; cell replacement therapy; cell type; experimental study; functional disability; human embryonic stem cell; in vivo; in vivo regeneration; induced pluripotent stem cell; innovation; loss of function; mdx mouse; motor function improvement; mouse model; muscle regeneration; myogenesis; novel; novel strategies; postnatal; programs; repaired; satellite cell; skeletal muscle wasting; socioeconomics; stem; stem cells; stemness; transcription factor

Muscle wasting, caused by aging, genetic mutations, cancan-associated cachexia, or traumatic injury, can result in significant functional impairment, and is a challenging clinical problem with a significant socioeconomic burden on our healthcare system. We have shown that functional myoblasts are readily derived from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), allowing us to begin to study Duchenne muscular dystrophy (DMD), the most common genetic disorder of muscle. However, we know little about i) how early myogenic events are genetically controlled during development, ii) whether embryonic PAX7 expressing myogenic stem/progenitor cells adopt postnatal `satellite-like' fate, and iii) how DMD is occurred in skeletal muscle stem/progenitor cell stage as well as their relevance for cell replacement therapy. First, using multiple genetic reporter lines to recapitulate human myogenic events, we will depict a time- course analysis of transcriptional landscape followed by `loss of function' analysis to address essential questions regarding which critical cell intrinsic/extrinsic component(s) govern the skeletal muscle specification process and stem cell maintenance. Secondly, by performing serial transplantation of human PAX7::GFP+ putative skeletal muscle stem/progenitor cells in mouse model, we will interrogate how the embryonic cells become to postnatal satellite-like cell fate. Thirdly, based on our observation on DYSTROPHIN expression in human skeletal muscle stem/progenitor cells, we will investigate stage-specific role(s) of DYSTROPHIN and its long intergenic non- coding RNAs (LincRNAs), in healthy and DMD condition. In addition, we will interrogate in vivo regeneration ability of patient-specific PAX7::GFP+ cells of genetically corrected DMD-hiPSC lines. Our proposed experiments are expected to expand and strengthen our current conception of myogenic specification events, and to accelerate a wide range of research on skeletal muscle disorders, e.g. traumatic muscle damages, genetic muscular dystrophies, neuromuscular diseases, type II diabetes and cancer-induced cachexia.

由衰老、基因突变、康康相关恶病质或外伤引起的肌肉萎缩， 可能导致严重的功能障碍，并且是一个具有挑战性的临床问题 我们的医疗保健系统的社会经济负担。 我们已经证明，功能性成肌细胞很容易源自人类胚胎干细胞 （hESC）和人类诱导多能干细胞（hiPSC），使我们能够开始研究杜氏肌细胞 营养不良（DMD），最常见的肌肉遗传性疾病。然而，我们对 i) 多久知之甚少 肌源性事件在发育过程中受到遗传控制，ii) 胚胎 PAX7 是否表达 肌源性干细胞/祖细胞采用出生后“卫星样”命运，以及 iii) DMD 是如何在骨骼中发生的 肌肉干/祖细胞阶段及其与细胞替代疗法的相关性。 首先，使用多个基因报告系来概括人类肌源性事件，我们将描述一个时间- 转录景观的过程分析，然后是“功能丧失”分析，以解决关键问题 关于哪些关键细胞内在/外在成分控制骨骼肌规格的问题 过程和干细胞维护。 其次，通过对人类 PAX7::GFP+ 假定骨骼肌进行连续移植 小鼠模型中的干/祖细胞，我们将探究胚胎细胞在出生后如何变化 类似卫星的细胞命运。 第三，基于我们对人体骨骼肌中肌营养不良蛋白表达的观察 干/祖细胞，我们将研究 DYSTROPHIN 的阶段特异性作用及其长基因间非- 健康和 DMD 状态下的编码 RNA (LincRNA)。此外，我们还将探讨体内再生 基因校正的 DMD-hiPSC 系的患者特异性 PAX7::GFP+ 细胞的能力。 我们提出的实验预计将扩大和加强我们目前的肌源性概念 规范事件，并加速骨骼肌疾病的广泛研究，例如创伤性的 肌肉损伤、遗传性肌营养不良、神经肌肉疾病、II 型糖尿病和癌症引起的 恶病质。

项目成果

Inhibition of the Combinatorial Signaling of Transforming Growth Factor-Beta and NOTCH Promotes Myotube Formation of Human Pluripotent Stem Cell-Derived Skeletal Muscle Progenitor Cells.

• DOI: 10.3390/cells10071649
• 发表时间: 2021-06-30
• 期刊: Cells
• 影响因子: 6
• 作者: Choi IY;Lim HT;Che YH;Lee G;Kim YJ
• 通讯作者: Kim YJ

Human pluripotent stem cell-derived myogenic progenitors undergo maturation to quiescent satellite cells upon engraftment.

人类多能干细胞衍生的肌源祖细胞在植入后成熟为静止卫星细胞。

• DOI: 10.1016/j.stem.2022.03.004
• 发表时间: 2022-04-07
• 期刊: CELL STEM CELL
• 影响因子: 23.9
• 作者: Sun, Congshan;Kannan, Suraj;Choi, In Young;Lim, HoTae;Zhang, Hao;Chen, Grace S.;Zhang, Nancy;Park, Seong-Hyun;Serra, Carlo;Iyer, Shama R.;Lloyd, Thomas E.;Lovering, Richard M.;Bin Lim, Su;Andersen, Peter;Wagner, Kathryn R.;Lee, Gabsang;Kwon, Chulan
• 通讯作者: Kwon, Chulan