Nuclear ryanodine receptor type 1: Role in skeletal muscle physiology and disease

核兰尼碱受体 1 型：在骨骼肌生理学和疾病中的作用

• 批准号: 8090729
• 负责人: Viktor Yarotskyy
• 金额: $ 9.61万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8090729
• 关键词: Advisory Committees; Area; Arrhythmia; Biochemistry; Biology; Biophysics; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium/calmodulin-dependent protein kinase; Cardiomyopathies; Cell Nucleus; Cell Proliferation; Central Core Myopathy; Characteristics; Commit; Complex; Coupled; Coupling; Defect; Development; Dihydropyridine Receptors; Disease; Disease model; Electrophysiology (science); Eligibility Determination; Environment; Exhibits; Fiber; Funding; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Hereditary Disease; Homeostasis; ITPR1 gene; Image; Inositol; Instruction; Knowledge; Laboratories; Leadership; Life; Link; Long-Term Effects; Malignant hyperpyrexia due to anesthesia; Mediating; Membrane; Mentored Research Scientist Development Award; Mentors; Molecular; Molecular Biology; Mus; Muscle; Muscle Fibers; Muscle function; Mutation; Myopathy; Nuclear; Nuclear Envelope; Nucleoplasm; Outcome; Paper; Pathway interactions; Peer Review; Pharmacology; Phenotype; Philosophy; Photometry; Physiological; Physiology; Play; Positioning Attribute; Process; Productivity; Property; Readiness; Regulation; Research; Research Personnel; Research Technics; Research Training; Reticulum; Role; RyR1; Ryanodine; Ryanodine Receptor Calcium Release Channel; Ryanodine Receptors; Sarcoplasmic Reticulum; Scientist; Seminal; Signal Pathway; Signal Transduction; Skeletal Muscle; Societies; Sudden Death; Technology; Testing; Thinking; Training; Training Programs; Transcriptional Regulation; Work; Writing; career; cell growth; cell type; design; disease phenotype; innovation; insight; interdisciplinary approach; member; mouse model; mutant; novel; oral communication; professor; programs; protein expression; protein kinase C kinase; residence; skills; transcription factor

DESCRIPTION (provided by applicant): The work outlined in this proposal is intended to fund an intensive mentored training program that promotes the development of the applicant into a complete and multi-disciplinarily trained independent investigator. The emphasis and strengths of the application are aligned according to three areas: 1. applicant credentials, 2. training environment, and 3. Innovative research. 1. Applicant Credentials. Starting a new position in Dr. Dirksen laboratory "aligned all stars in the sky" for Dr. Yarotskyy by creating ideal conditions for the proposed Mentored Research Scientist Development Award (K01) application: proven track-record of productivity, eligibility for program (obtaining permanent residence status), readiness to establish scientific independence, change in research direction, and exceptional technical and professional training opportunity. The applicant has outstanding skills and great expertise in electrophysiology and biophysics, but lacks essential in-depth training in other key components required for a successful independent career in academic research. The PI's short-term career goals include expanding scope of technical skills, begin to develop and establish an independent line of research, and to refine essential professional skills (including scientific writing, oral communication, instruction, trainee mentoring, and involvement in peer-review). Long-term goals include obtaining a tenure-track position, establishing an independent line of research, training future scientists, emerging as leader in chosen field of research field, and participation in professional and society leadership. This proposal is explicitly designed to: 1. Develop a new area of research for applicant, 2. provide needed additional technical training and expertise, 3. promote continued development of leadership, mentoring and professional skills, and 4. establish laboratory and project management skills. Completing the proposed training will achieve all short-term goals and provide expertise required to accomplish the applicant's long-term goals. 2. Training Environment. Dr. Dirksen is fully committed to the complete training of outstanding and bright young scientists. Dr. Dirksen's laboratory is an ideal place for the applicant's proposed training and first independent step to take place. The Dirksen laboratory employs a broad range of cutting-edge technologies routinely used by the research team with diverse expertise in imaging, photometry, molecular biology and biochemistry, adenoviral technology, mouse genetics and disease models. Dr. Dirksen's overall philosophy is to provide all trainees with an enriched and nurturing scientific environment that promotes trainee development into careful, critical-thinking problem-solvers and effective scientific communicators. Dr. Yarotskyy will benefit from intense training in new research techniques, mentoring, leadership, and professional skills. Being a proud member of the Department of Physiology and Pharmacology, Dr. Yarotskyy will also benefit from great support from other well-established senior investigators. Overall oversight of the training program will be closely supervised by an outstanding Mentoring Advisory Committee consisting of Drs. R. Dirksen, D. Yule and S.-S. Sheu. Toward the end of the K01 award, Dr. Yarotskyy will use results obtained during this project to justify and extend the scope of the project by formulating an independent New Investigator R01 application that will enable him to launch his independent research program and obtain a tenure track Assistant Professor position within the department. 3. Innovative Research. The central hypothesis to be tested here is that: "Nuclear RyR1 (nRyR1) release channels are important regulators of nucleoplasmic Ca2+ signaling and gene regulation in skeletal muscle." The type I ryanodine receptor (RyR1) plays an essential role in skeletal muscle function. Mutations in RyR1 result in several debilitating and life-threatening myopathies, including malignant hyperthermia (MH) and central core disease (CCD). The molecular mechanisms underlying CCD core formation have yet to be fully clarified. The current paradigm is that mutant RyR1 channels in the sarcoplasmic reticulum (SR) alter cytoplasmic Ca2+ signaling that eventually leads to the MH and CCD phenotypes. We challenge this paradigm by proposing a role for mutant RyR1 channels in the nucleoplasmic reticulum (nRyR1) that result in aberrant nucleoplasmic Ca2+ signaling and altered transcription factor localization and activity that underlie the formation of cores and fiber type changes characteristic of CCD. In this project, we propose that: 1. nRyR1 regulates nucleoplasmic Ca2+ transients by controlling Ca2+ release from the perinuclear space during stimulation; 2. nucleoplasmic Ca2+ impacts NFAT localization and activity which underlies fiber type specification; 3. morphological changes in CCD result from altered gene expression driven by aberrant nRyR1 channel activity. This project will employ a multidisciplinary approach, including confocal imaging, biochemistry, molecular biology, and electrophysiology to test the central hypothesis. This project will test the validity of a novel pathophysiological mechanism by which altered RyR1 channel in impacts skeletal muscle disease.

描述（由申请人提供）：本提案中概述的工作旨在资助一项强化指导培训计划，以促进申请人发展成为一名完整的、经过多学科培训的独立研究者。申请的重点和优势根据三个领域进行调整：1. 申请人资质，2. 培训环境，3. 创新研究。 1. 申请人的凭据。在德克森博士实验室开始一个新职位，为 Yarotskyy 博士“排列了天空中的所有星星”，为拟定的导师研究科学家发展奖 (K01) 申请创造了理想的条件：良好的生产力记录、项目资格（获得永久居留身份）、准备建立科学独立性、研究方向的改变以及特殊的技术和专业培训机会。申请人在电生理学和生物物理学方面拥有出色的技能和丰富的专业知识，但缺乏成功的独立学术研究生涯所需的其他关键组成部分的必要深入培训。 PI的短期职业目标包括扩大技术技能范围，开始开发和建立独立的研究路线，并完善基本的专业技能（包括科学写作、口头交流、指导、实习生指导和参与同行评审）。长期目标包括获得终身教授职位、建立独立的研究路线、培养未来的科学家、成为所选研究领域的领导者以及参与专业和社会领导。该提案的明确目的是： 1. 为申请人开发新的研究领域， 2. 提供所需的额外技术培训和专业知识， 3. 促进领导力、指导和专业技能的持续发展，以及 4. 建立实验室和项目管理技能。完成拟议的培训将实现所有短期目标，并提供实现申请人长期目标所需的专业知识。 2. 培训环境。德克森博士完全致力于对杰出、聪明的年轻科学家的全面培养。德克森博士的实验室是申请人进行拟议培训和迈出独立第一步的理想场所。德克森实验室采用了研究团队常用的广泛尖端技术，这些技术在成像、光度测定、分子生物学和生物化学、腺病毒技术、小鼠遗传学和疾病模型等方面拥有不同的专业知识。德克森博士的总体理念是为所有学员提供丰富和培育的科学环境，促进学员发展成为细心、批判性思维的问题解决者和有效的科学传播者。 Yarotskyy 博士将受益于新研究技术、指导、领导力和专业技能方面的强化培训。作为生理学和药理学系的一名自豪的成员，Yarotskyy 博士还将受益于其他知名高级研究人员的大力支持。培训计划的总体监督将由由博士组成的杰出指导咨询委员会密切监督。 R. Dirksen、D. Yule 和 S.-S。谢乌。在 K01 奖结束时，Yarotskyy 博士将利用该项目期间获得的结果，通过制定独立的新研究者 R01 申请来证明和扩展该项目的范围，这将使他能够启动他的独立研究计划，并获得系内的终身教授助理教授职位。 3.创新研究。这里要测试的中心假设是：“核 RyR1 (nRyR1) 释放通道是骨骼肌中核质 Ca2+ 信号传导和基因调控的重要调节因子。” I 型兰尼碱受体 (RyR1) 在骨骼肌功能中发挥着重要作用。 RyR1 突变会导致多种使人衰弱和危及生命的肌病，包括恶性高热 (MH) 和中枢性核心疾病 (CCD)。 CCD 核心形成的分子机制尚未完全阐明。当前的范例是肌浆网 (SR) 中的突变 RyR1 通道改变细胞质 Ca2+ 信号传导，最终导致 MH 和 CCD 表型。我们通过提出突变型 RyR1 通道在核质网 (nRyR1) 中的作用来挑战这一范式，这种作用会导致异常的核质 Ca2+ 信号传导以及转录因子定位和活性的改变，这些是 CCD 核心形成和纤维类型变化特征的基础。在这个项目中，我们提出： 1. nRyR1 通过控制刺激过程中核周间隙 Ca2+ 的释放来调节核质 Ca2+ 瞬变； 2. 核质 Ca2+ 影响 NFAT 定位和活性，这是纤维类型规范的基础； 3. CCD 的形态变化是由异常的 nRyR1 通道活性驱动的基因表达改变引起的。该项目将采用多学科方法，包括共焦成像、生物化学、分子生物学和电生理学来检验中心假设。该项目将测试一种新的病理生理学机制的有效性，通过改变 RyR1 通道来影响骨骼肌疾病。