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：本项目的目的是为Silkenstein提供研究和职业发展培训 Sidhom是佛罗里达大学的博士生。这份行政补充文件是对PA- 21-071：“研究补充，以促进健康相关研究的多样性（行政补充）”将促进 候选人的职业发展，以及有助于母项目的使命-定义 生物钟下游的分子靶点调节肌肉结构， 实力 Sidhom女士提议的研究的主要目标将是进行与2010年目标2有关的实验。 家长补助金首先，她将学习使用共聚焦成像方法与肌节抗体，以确定 肌节的M线和Z线结构元件与对照组和肌肉特异性 Bmal 1 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，以测试其是否足以恢复结构和肌肉力量。Sidhom女士会积极地 与柯林道格拉斯，另一个博士生工作的家长补助金以及克里斯托弗沃尔夫博士，一个 这个项目的博士后助理R 01的PI Esser博士将成为Sidhom女士的主要导师 在佛罗里达大学的医学院内。我们已经建立了她的委员会， 包括：Russ Hepple，Christiaan Leeuwenburgh和Richard Dunn博士。他们对女士很热情。 Sidhom作为一名学生，并提供骨骼肌和成像方法的适当专业知识。Silvana 已经完成了第一年的必修课程，因此委员会将与Sidhom女士合作， 一系列教学课程，包括每年秋季教授的骨骼肌课程。委员会还将帮助 指导Silencia进行体验式培训活动，使她具备必要的技能， 独立的科学家，并履行国家卫生研究院的使命，以促进科学家的多元化人口的职业生涯。 该项目与母赠款的目标2直接相关： 具体目标2：检测时钟控制基因Rbm 20和/或Tcap在肌节中的作用 结构和肌肉功能。该目的将使用AAV递送Tcap和/或Rbm 20到Bmal 1缺陷型小鼠中。 肌肉，以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。 肌节结构要素将通过超分辨率显微镜与反卷积技术获得 用抗体定义2D和3D特性，包括肌节长度、粗丝中心性、M带 和Z线完整性。将对Rbm 20肌肉进行深度RNA测序，以确定剪接 变化功能结局将包括体外测量，以测试结构变化对两者的影响 被动/刚度和主动/最大力和收缩率特性。