The interaction between mechanical forces and cytoskeletal impairments in podocyte mediated kidney disease

足细胞介导的肾病中机械力与细胞骨架损伤之间的相互作用

基本信息

批准号：
10225541
负责人：
Di Feng
金额：
$ 15.07万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10225541
关键词：
Actinin Actins Affect Albuminuria Aleurites Antihypertensive Agents Apoptosis Applications Grants Award Biomechanics Biophysics Blood flow Boston Bowman&apos s space CRISPR/Cas technology Caliber Cell Death Chronic Kidney Failure Contracts Cytoskeleton DNA Sequence Alteration Defect Discipline Disease Doctor of Philosophy Educational workshop Environment Event Exhibits Failure Fellowship Fibroblasts Filtration Focal Adhesions Focal Segmental Glomerulosclerosis Foundations Functional disorder Genetic Genetic study Glomerular Capillary Goals Human Image Immunosuppression Impairment In Vitro Industry Injury Injury to Kidney International Kidney Diseases Knock-in Knowledge Lead Learning Mass Spectrum Analysis Measures Mechanical Stress Mediating Mentors Mentorship Methods Modeling Molecular Mus Mutation Nature Other Genetics Pathogenesis Pathway interactions Patients Periodicity Phosphorylation Pluripotent Stem Cells Post-Translational Protein Processing Proteins Rattus Research Research Personnel Research Training Resolution Resources Seminal Signal Pathway Site Stress Stretching Techniques Technology Time Traction Force Microscopy Training Programs Wisconsin Work alpha Actinin base blood filter career development crosslink design effective therapy efficacy evaluation experience glomerular basement membrane glomerular filtration in vivo intravital microscopy kidney cell mechanical force medical schools mouse model multidisciplinary mutant non-genetic novel novel therapeutics organ on a chip personalized medicine podocyte protein crosslink reconstitution response shear stress simulation skills targeted treatment

项目摘要

Project Summary/Abstract This K01 grant proposal describes a five-year mentored training program designed to transition Dr. Di Feng to become an independent academic investigator. Dr. Feng obtained her Ph.D. at the Medical College of Wisconsin under the mentorship of Dr. Allen Cowley. She is now completing her postdoctoral fellowship in the lab of Dr. Martin Pollak, an international leader in studying the genetics of glomerular kidney disease. Dr. Feng has focused her research on elucidating the mechanism by which mutations in ACTN4 – an important cytoskeleton protein – lead to a form of glomerular kidney disease called focal segmental glomerulosclerosis (FSGS). The inability to better characterize the podocyte dysfunction that underlies FSGS has hindered the field in establishing more specific, personalized treatments beyond broad immunosuppression and anti- hypertensive therapy. Dr. Feng has focused her research on the mutant podocyte’s response to the mechanical stresses it experiences while filtering blood flow in the glomerulus. She has so far shown that the biophysical changes conferred by disease-causing mutant ACTN4 render the podocyte brittle, exhibiting failure of contractile forces and actin cytoskeleton disruption in response to periodic stretch. In the current proposal, Aim 1 seeks to further define the impaired response of human podocytes caused by mutant ACTN4, not only to stretch but also to shear stress. She will employ organ-on-a-chip methods to better simulate these stresses while quantifying the associated biomechanical and molecular responses of podocytes. Aim 2 will determine whether post-translational phosphorylation of ACTN4 also impairs the response of podocytes to mechanical stress, using mouse models and biomechanical studies of podocytes isolated from these mice. Aim 3 plans to use CRISPR/Cas technology to generate a mutant ACTN4 rat model and use intravital microscopy to measure the in vivo mechanical stresses within mutant and WT glomeruli. Through the proposed research, she will learn organ-on-a-chip methods, mass spectrometry, super-resolution imaging, and intravital microscopy. She has assembled a team of mentors and advisors under Dr. Pollak entailing leaders in these respective disciplines, including Dr. Donald Ingber, Dr. Bruce Molitoris, Dr. Hanno Steen, Dr. Douglas Richardson, as well as Dr. Roger Tung, who will provide advice related to the translational value of her work. Dr. Feng will spend 95% of her time under this award toward the proposed research, and her training plan includes didactic courses, seminars, and career development workshops at Harvard. The proposed project will make Dr. Feng competitive for independent research awards, for which she plans to apply her findings from ACTN4 and the above multidisciplinary methods to further study how defects in the actin-based cytoskeleton impair the podocyte’s response to the mechanical stresses experienced in vivo. The advancement of her goals will take place within Harvard’s vast resources and connections to thought-leaders, situated in the unique environment of Boston that integrates academics and industry.

项目概要/摘要这项 K01 拨款提案描述了一项为期五年的指导培训计划，旨在将冯迪博士转变为冯博士在美国医学院获得博士学位。在艾伦·考利博士的指导下，她正在威斯康星州完成她的博士后研究。 Martin Pollak 博士是肾小球肾病遗传学研究领域的国际领军人物冯博士的实验室。她的研究重点是阐明 ACTN4 突变的机制——一个重要的细胞骨架蛋白——导致一种称为局灶节段性肾小球硬化症的肾小球疾病（FSGS）。无法更好地表征 FSGS 背后的足细胞功能障碍阻碍了研究。建立超越广泛的免疫抑制和抗病毒治疗的更具体、个性化的治疗方法冯博士的研究重点是突变足细胞对高血压的反应。迄今为止，她已经证明了肾小球在过滤血流时所经历的机械应力。致病突变体 ACTN4 带来的生物物理变化使足细胞变得脆弱，表现出衰竭在当前的提议中，收缩力和肌动蛋白细胞骨架破坏响应周期性拉伸。目标 1 旨在进一步定义突变 ACTN4 引起的人类足细胞反应受损，不仅是为了她将采用芯片上器官的方法来更好地模拟这些应力。目标 2 将确定同时量化足细胞的相关生物力学和分子反应。 ACTN4 的翻译后磷酸化是否也会损害足细胞对机械的反应 Aim 3 计划利用小鼠模型并对从这些小鼠中分离出的足细胞进行生物力学研究。利用CRISPR/Cas技术生成突变ACTN4大鼠模型并利用活体显微镜进行测量通过拟议的研究，她将了解突变体和 WT 肾小球内的体内机械应力。器官芯片方法、质谱、超分辨率成像和活体显微镜。在 Pollak 博士的领导下组建了一支由导师和顾问组成的团队，其中包括这些各自学科的领导者，包括 Donald Ingber 博士、Bruce Molitoris 博士、Hanno Steen 博士、Douglas Richardson 博士以及 Roger Tung 将花费 95% 的资金，为她的工作的转化价值提供建议。她在该奖项下用于拟议研究的时间，以及她的培训计划包括教学课程，拟议的项目将使冯博士在哈佛大学举办研讨会和职业发展研讨会。她计划将自己在 ACTN4 和上述多学科方法进一步研究基于肌动蛋白的细胞骨架的缺陷如何损害足细胞对体内机械应力的反应将需要她的目标的进展。哈佛拥有丰富的资源和与思想领袖的联系，坐落在独特的环境中波士顿大学集学术与工业于一体。