Muscle clock and weakness: diversity supplement

肌肉时钟和弱点：多样性补充

• 批准号: 10414186
• 负责人: Karyn A Esser
• 金额: $ 3.73万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-29 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10414186
• 关键词: 3-Dimensional; ARNTL gene; ATAC-seq; Administrative Supplement; Age; Aging; Antibodies; Attention; Bioluminescence; ChIP-seq; Chromatin; Circadian Rhythms; Coupled; DNA Binding; Data; Dose; Elements; Female; Florida; Gene Expression; Genes; Genetic; Genomics; Health; Knockout Mice; Learning; Length; Longevity; Measures; Medicine; Mentors; Microscopy; Mission; Molecular; Molecular Target; Muscle; Muscle Weakness; Muscle function; Outcome; Output; Phenotype; Physiological; Population Heterogeneity; Property; Proteins; RNA Splicing; Regulation; Research; Resolution; Rodent Model; Role; Sarcomeres; Scientist; Series; Skeletal Muscle; Structure; Students; Techniques; Testing; Thick Filament; Tissues; Training; Training Activity; United States National Institutes of Health; Universities; Work; adeno-associated viral vector; bone imaging; career; career development; circadian pacemaker; college; confocal imaging; doctoral student; epigenomics; experience; experimental study; falls; functional outcomes; graduate student; human subject; imaging approach; improved; interest; male; metabolomics; mouse model; muscle strength; muscular structure; parent grant; parent project; pre-clinical; research and development; response; skills; transcriptome sequencing; transcriptomics

Abstract: The purpose of this project is to provide research and career development training for Silvana Sidhom, a current PhD student at the University of Florida. This administrative supplement in response to PA- 21-071: “Research Supplements to Promote Diversity in Health-Related Research (Admin Supp)” will facilitate career development for the candidate and as well as contribute to the mission of the parent project – to define molecular targets downstream of the circadian clock that modulate muscle structure with implications for strength. The primary objectives of Ms. Sidhom’s proposed research will be to work on experiments related to Aim 2 of the parent grant. First, she will learn to use confocal imaging approaches with sarcomeric antibodies to define changes in the M-line and Z-line structural elements of the sarcomere from the control and muscle specific Bmal1 KO mice. Second, she will use AAV approaches to restore expression of selected sarcomeric genes, such as Tcap, to test if that is sufficient to restore structure and muscle force. Ms. Sidhom will work actively with Collin Douglas, another PhD student working on the parent grant as well as Dr. Christopher Wolff, a postdoctoral associate on this project. Dr. Esser, the PI of the R01, will be the primary mentor for Ms. Sidhom within the College of Medicine at the University of Florida. We have established her committee and this includes: Drs. Russ Hepple, Christiaan Leeuwenburgh and Richard Dunn. They are enthusiastic about Ms. Sidhom as a student and provide appropriate expertise in skeletal muscle and imaging approaches. Silvana has completed her first year of required courses so the committee will work with Ms. Sidhom to outlined a series of didactic courses, including the skeletal muscle course taught each fall. The committee will also help guide Silvana with experiential training activities that will equip her with the necessary skills to progress as an independent scientist and to fulfill the NIH mission to advance the careers of a diverse population of scientists. The project is directly related to Aim 2 of the parent grant: Specific Aim 2: To test the clock controlled genes, Rbm20 and/or Tcap, for their roles in sarcomere structure and muscle function. This aim will use AAV delivery of Tcap and/or Rbm20 in Bmal1 deficient muscle to test their contribution to sarcomere structure, myofibrillar protein composition and muscle function. Sarcomere structural elements will be obtained by super-resolution microscopy with deconvolution techniques with antibodies to define 2D and 3D properties including sarcomere length, thick filament centrality, M-band and Z line integrity. Deep RNA-sequencing will be performed on the Rbm20 muscles to define splicing changes. Functional outcomes will include ex vivo measures to test the impact of structural changes on both passive/stiffness and active/max force and rate of contraction properties.

摘要：该项目的目的是为 Silvana 提供研究和职业发展培训 Sidhom，佛罗里达大学现任博士生。本行政补充是为了回应 PA- 21-071：“促进健康相关研究多样性的研究补充（行政补充）”将促进 候选人的职业发展，并为父项目的使命做出贡献——定义 生物钟下游的分子目标调节肌肉结构，对 力量。 Sidhom 女士提议的研究的主要目标是进行与目标 2 相关的实验 家长补助金。首先，她将学习使用共聚焦成像方法和肌节抗体来定义 肌节 M 线和 Z 线结构元件的变化（来自控制和肌肉特异性） Bmal1 KO 小鼠。其次，她将使用 AAV 方法来恢复选定的肌节基因的表达， 例如 Tcap，以测试是否足以恢复结构和肌肉力量。 Sidhom女士将积极工作 与另一位从事家长补助金的博士生科林·道格拉斯 (Collin Douglas) 以及克里斯托弗·沃尔夫 (Christopher Wolff) 博士 该项目的博士后助理。 R01 的 PI Esser 博士将成为 Sidhom 女士的主要导师 在佛罗里达大学医学院内。我们已经成立了她的委员会 包括：博士。拉斯·赫普尔、克里斯蒂安·列文堡和理查德·邓恩。他们对小姐姐很热情。 Sidhom 作为一名学生，提供骨骼肌和成像方法方面的适当专业知识。西尔瓦娜 已完成第一年的必修课程，因此委员会将与 Sidhom 女士合作概述 系列教学课程，包括每年秋季教授的骨骼肌课程。组委会也将提供帮助 指导西尔瓦娜进行体验式培训活动，使她具备作为一名进步人士所需的技能 独立科学家并履行 NIH 的使命，促进多元化科学家的职业生涯。 该项目与母基金的目标 2 直接相关： 具体目标 2：测试时钟控制基因 Rbm20 和/或 Tcap 在肌节中的作用 结构和肌肉功能。该目标将在 Bmal1 缺陷中使用 AAV 递送 Tcap 和/或 Rbm20 肌肉来测试它们对肌节结构、肌原纤维蛋白组成和肌肉功能的贡献。 肌节结构元素将通过超分辨率显微镜和反卷积技术获得 使用抗体定义 2D 和 3D 属性，包括肌节长度、粗丝中心性、M 带 和 Z 线完整性。将在 Rbm20 肌肉上进行深度 RNA 测序以确定剪接 变化。功能结果将包括离体测量，以测试结构变化对两者的影响 被动/刚度和主动/最大力和收缩率属性。