Regulation of Mitsugumin 29 expression in muscle physiology and diseases

Mitsugumin 29 表达在肌肉生理学和疾病中的调节

• 批准号: 9028620
• 负责人: Hua Zhu
• 金额: $ 32.99万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9028620
• 关键词: 3' Untranslated Regions; Ablation; Accounting; Address; Animal Model; Appearance; Binding Sites; Biochemical; Biogenesis; Biological Assay; Biological Process; Biopsy; Cell Culture Techniques; Cell Line; Cells; Coupling; DYSF gene; Data; Development; Direct Lytic Factors; Disease; Down-Regulation; Electron Microscopy; Exhibits; Gene Delivery; Genes; Genetic; Goals; Human; Immunoblotting; Immunohistochemistry; In Vitro; Injury; Link; Maintenance; Mediating; Membrane; MicroRNAs; Modeling; Molecular; Mus; Muscle; Muscle Fibers; Muscular Dystrophies; Mutagenesis; Myoblasts; Myopathy; Names; Patients; Physiological; Physiology; Play; Post-Transcriptional Regulation; Principal Investigator; Process; Protein Family; Proteins; RNA; Regulation; Research; Research Design; Research Personnel; Role; Sarcolemma; Signal Transduction; Skeletal Muscle; Staging; Stress; Structure; Synaptophysin; System; Testing; Therapeutic; Transgenic Animals; Transgenic Mice; Tubular formation; improved; in vivo; insight; mdx mouse; member; mouse model; muscle regeneration; muscle strength; muscular dystrophy mouse model; professor; public health relevance; regenerative; research study; tool

 DESCRIPTION (provided by applicant): Mitsugumin 29 (MG29) is a muscle-specific member of the synaptophysin family proteins that participates in controlling the maturation and development of the transverse-tubule (TT) structure and the maintenance of intracellular Ca2+ signaling in skeletal muscle. Genetic ablation of mg29 leads to defective TT structure in skeletal muscle, which show similarity to the abnormal TT network observed in animal models of, and human patients with, muscular dystrophy. The principal investigator of this application has obtained evidence that revealed an essential role for MG29 in controlling the TT membrane network in muscle physiology and diseases. Immunoblot showed that expression of MG29 was reduced in both human and mice with muscular dystrophy, indicating the possibility that MG29 may be involved in the development of muscular dystrophy. Using biochemical assay, we identified a functional interaction between MG29, dysferlin and Bin1, other TT- resident proteins with important roles in muscular dystrophy. Toward understanding the biological function for MG29 in control of TT biogenesis, we used the HMCL-7304 myoblast cell line derived from human muscle. Cultured myotubes derived from HMCL-7304 cells do not express MG29 and lack TT network. Preliminary study with rescue of MG29 in HMCL-7304 myotubes revealed the initiation of TT membranes. Establishing an in vitro system with control of TT membrane can potentially be a useful tool for muscle physiology research. The mg29 gene contains a unique 3' untranslated region (UTR) with potential binding sites for microRNA (miRNA) or RNA-stabilizing factors. Mutagenesis studies revealed that miR-181a could target a region in the 3'UTR that is highly conserved between mouse and human mg29 genes to exhibit a strong control of MG29 expression in skeletal muscle. Studies from other investigators have shown that miR-181 was significantly elevated in human patients with muscular dystrophy. These data suggest an intriguing possibility that elevated miR-181a may be a contributing factor for the reduced MG29 expression under muscular dystrophic conditions. Experiments outlined in this proposal will focus on testing the hypothesis that "MG29 is an integral component of TT biogenesis in skeletal muscle, and miRNA-mediated control of MG29 expression contributes to changes in TT integrity during muscular dystrophy". First, by using in vitro cell culture and in vivo transgenic animal approaches, we will discern the biological function for MG29 in control of TT integrity and remodeling in muscle physiology and dystrophy. Second, we will elucidate the mechanisms that underlie miRNA-mediated control of MG29 expression in skeletal muscle, and to identify the mechanisms that contribute to reduction of MG29 in dystrophic muscle. Using AAV-mediated gene delivery of antagomir into the mdx mice, we will test whether suppression of miRNA-mediated down-regulation of MG29 expression can improve TT structure and rescue muscle strength in dystrophic muscle.

 描述（由申请人提供）：Mitsugumin 29 (MG29) 是突触素家族蛋白的肌肉特异性成员，参与控制横管 (TT) 结构的成熟和发育以及骨骼肌中细胞内 Ca2+ 信号传导的维持mg29 的基因消融导致骨骼肌中的 TT 结构缺陷，这与在动物模型和人类中观察到的异常 TT 网络相似该申请的主要研究人员获得的证据表明 MG29 在控制肌肉生理学和疾病中的 TT 膜网络中发挥着重要作用。免疫印迹显示 MG29 的表达在患有肌营养不良症的人和小鼠中均减少。 ，表明 MG29 可能参与肌营养不良症的发展，使用生化测定，我们鉴定了 MG29、dysferlin 和 Bin1（其他具有重要作用的 TT 驻留蛋白）之间的功能相互作用。为了了解 MG29 在控制 TT 生物发生中的生物学功能，我们使用了源自人类肌肉的 HMCL-7304 成肌细胞系，该细胞系不表达 MG29 并且缺乏 TT 网络。在 HMCL-7304 肌管中拯救 MG29 的研究揭示了 TT 膜的启动建立体外系统。控制 TT 膜可能成为肌肉生理学研究的有用工具，mg29 基因包含一个独特的 3' 非翻译区 (UTR)，具有 microRNA (miRNA) 或 RNA 稳定因子的潜在结合位点。 181a 可以靶向小鼠和人类 mg29 基因之间高度保守的 3'UTR 区域，以表现出对骨骼肌中 MG29 表达的强大控制。其他研究人员的研究表明。 miR-181 在患有肌营养不良症的人类患者中显着升高，这些数据表明一个有趣的可能性，即 miR-181a 可能是肌营养不良症条件下 MG29 表达减少的促成因素。 “MG29 是骨骼肌 TT 生物发生的一个组成部分，miRNA 介导的 MG29 表达控制有助于肌营养不良症期间 TT 完整性的变化”。通过体外细胞培养和体内转基因动物方法，我们将了解 MG29 在肌肉生理学和营养不良中控制 TT 完整性和重塑的生物学功能。其次，我们将阐明 miRNA 介导的骨骼肌中 MG29 表达控制的机制。 ，并确定有助于减少营养不良性肌肉中 MG29 的机制，使用 AAV 介导的 antagomir 基因递送至 mdx 小鼠中，我们将测试是否抑制 MG29。 miRNA 介导的 MG29 表达下调可以改善营养不良性肌肉的 TT 结构并挽救肌肉力量。