Alternative Splicing Regulation and Mechanotransduction in Skeletal Muscle

骨骼肌中的选择性剪接调节和机械转导

基本信息

批准号：
10403954
负责人：
Emma Hinkle
金额：
$ 2.84万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2022-11-18
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10403954
关键词：
Actins Adult Affect Alternative Splicing Antisense Oligonucleotides Atomic Force Microscopy Binding Biochemical Bioinformatics Biology Cells Cellular biology Co-Immunoprecipitations Code Communication Communities Complex Coupling Critical Thinking Data Defect Development Developmental Gene Doctor of Philosophy Education Educational process of instructing Embryo Environment Exhibits Exons Fellowship Generations Genes Genetic Genetic Transcription Goals Growth Human Individual Knockout Mice Lead Life Measures Membrane Mentors Mentorship Microfilaments Microscopy Molecular Movement Muscle Muscle Cells Muscle Contraction Muscle Development Muscle Fibers Muscle Proteins Muscular Atrophy Muscular Dystrophies Myopathy North Carolina Nuclear Translocation Nucleotides Pathologic Pathway interactions Periodicity Persons Phosphorylation Physics Physiological Play Polypyrimidine Tract-Binding Protein Positioning Attribute Process Protein Isoforms Proteins Public Health Publishing RNA RNA Processing RNA Splicing RNA-Binding Proteins Regulation Research Research Personnel Role Sarcomeres Signal Transduction Skeletal Muscle Solid Stretching Striated Muscles Testing Time Tissues Training Universities Variant Western Blotting Writing actin capping protein career collaborative environment extracellular fetal insight interdisciplinary approach mRNA Precursor mechanical properties mechanical stimulus mechanotransduction multidisciplinary muscle physiology muscular structure myogenesis novel programs recruit response skills trafficking transcriptome sequencing transmission process

项目摘要

PROJECT SUMMARY/ABSTRACT Skeletal muscle allows for controlled movement and fine-tuned coordination throughout the entire life of an individual. In all cells, the response to physical forces is critical; however, this is particularly important in skeletal muscles to facilitate movement, contraction, and force generation. Mechanotransduction allows cells to sense and respond to the external environment. In muscle cells, forces transmit through the Z-disc which are interspersed between sarcomeres and anchor actin filaments. In numerous muscular diseases and myopathies, mechanotransduction is altered and leads to loss of force, muscle wasting, and increased stiffness. At the molecular level, these pathological alterations are accompanied by extensive transcriptional changes and mis- regulation of alternative splicing, an RNA processing mechanism that allows single genes to code for multiple protein isoforms. A unique feature of skeletal muscle is that it exhibits one of the highest levels of tissue-specific and evolutionarily conserved alternative splicing. During muscle development, extensive alternative splicing changes occur to facilitate maturation of the tissue. Interestingly, numerous genes developmentally regulated by splicing encode proteins that are involved in membrane trafficking and localize to the sarcomere. One of these genes encodes the Capping Actin Protein of Muscle Z-Line Subunit Beta (CAPZB) protein. Besides its critical function in capping actin, CAPZB plays unconventional roles in sarcomere organization. Surprisingly, how mechanotransduction and alternative splicing are interconnected in muscles has not been deeply investigated and can reveal new insights about muscle diseases. In my proposal, I hypothesize that alternative splicing regulation contributes to the development of the mechanical properties of skeletal muscle. I will test this hypothesis in two specific aims. In aim 1, I will identify the role of two splicing regulators, the poly-pyrimidine tract binding protein 1 (PTBP1) and quaking protein (QK), in controlling the mechanical properties of muscle cells by stretching cells and investigating effects on mechanosensitive pathways. In aim 2, I will determine how CAPZB and its splice forms contribute to the mechanosensitivity of muscle cells by using force microscopy and functional studies. My long-term goal is to be an independent scientific leader who can lead a team. Therefore, the training I will receive through this fellowship will facilitate my growth in becoming a muscle biologist with expertise in RNA processing, and solid skills in mentorship, writing and teaching. My sponsor and co-sponsor are experts in muscle physiology, cell biology, and, alternative splicing and I have recruited collaborators and mentors with expertise in physics and mechanotransduction to facilitate a multidisciplinary dissertation. All of them are strongly committed to mentoring and education and will support me during my Ph.D. and as I move forward in my career. Finally, the University of North Carolina at Chapel Hill has strong communities of RNA Biology, mechanotransduction, and membrane trafficking that contribute to the collaborative environment I am part of.

项目摘要/摘要骨骼肌可以在整个生命中进行控制和微调的协调个人。在所有细胞中，对物理力的反应至关重要。但是，这在骨骼中尤其重要肌肉促进运动，收缩和力量产生。机械转移使细胞感知并回应外部环境。在肌肉细胞中，力通过z二轴传播散布在肉瘤和锚固肌动蛋白丝之间。在许多肌肉疾病和肌病中，机械转导改变并导致力损失，肌肉浪费和增加的刚度。在分子水平，这些病理改变伴随着广泛的转录变化和错误调节替代剪接的调节，一种RNA处理机制，允许单个基因代码多个蛋白质同工型。骨骼肌的独特特征是它表现出最高水平的组织特异性和进化保守的替代剪接。在肌肉发育期间，发生了广泛的替代剪接变化，以促进组织。有趣的是，通过剪接编码蛋白质在发育中发育中的许多基因在膜贩运中，本地化到肉皮。这些基因之一编码粘蛋白蛋白上限肌肉Z线亚基β（CAPZB）蛋白的蛋白质。除了它在封闭肌动蛋白方面的关键功能外，CAPZB播放肌节组织中非常规角色。令人惊讶的是，机械转移和替代方式如何在肌肉中互连的剪接尚未进行深入研究，可以揭示有关有关的新见解肌肉疾病。在我的提议中，我假设替代拼接法规有助于骨骼肌机械性能的发展。我将以两个具体的目的来检验这一假设。在 AIM 1，我将确定两个剪接调节剂的作用，即多吡啶氨酸裂纹结合蛋白1（PTBP1）和 Quaking蛋白（QK），通过拉伸细胞和研究对机械敏感途径的影响。在AIM 2中，我将确定CAPZB及其剪接形式如何通过使用力显微镜和功能研究来促进肌肉细胞的机械敏感性。我的长期目标是成为可以领导团队的独立科学领袖。因此，培训我将通过这项奖学金获得，将有助于成为具有专业知识的肌肉生物学家的成长 RNA处理以及指导，写作和教学方面的扎实技能。我的赞助商和共同赞助商是专家在肌肉生理学，细胞生物学以及替代剪接方面，我已经招募了合作者和导师物理和机械转导专业知识，以促进多学科论文。他们都是强烈致力于指导和教育，并将在我的博士学位上支持我。当我前进我的职业。最后，北卡罗来纳大学教堂山分校拥有牢固的RNA生物学社区，机械转移和膜贩运有助于我的合作环境。