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

• 批准号: 9750298
• 负责人: Di Feng
• 金额: $ 14.53万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 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/9750298
• 关键词: Actinin; Actins; Affect; Albuminuria; Aleurites; Antihypertensive Agents; Apoptosis; Applications Grants; Award; Biomechanics; Biophysics; Blood flow; Boston; Bowman' 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; International; Kidney; 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; 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

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 成为一名独立的学术研究者。冯博士获得博士学位。在医学院 威斯康星州在艾伦·考利博士的指导下。她现在正在完成她的博士后研究金 Martin Pollak 博士的实验室是研究肾小球肾病遗传学的国际领先者。冯博士 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 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 高血压治疗。冯博士的研究重点是突变足细胞对 它在过滤肾小球中的血流时承受机械应力。迄今为止，她已经表明 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 响应周期性拉伸的收缩力和肌动蛋白细胞骨架破坏。在目前的提案中， 目标 1 旨在进一步明确突变 ACTN4 引起的人类足细胞反应受损，不仅是为了 不仅可以拉伸，还可以承受剪切应力。她将采用芯片器官方法来更好地模拟这些压力 同时量化足细胞的相关生物力学和分子反应。目标 2 将决定 ACTN4 的翻译后磷酸化是否也会损害足细胞对机械的反应 使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。目标3计划 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 突变体和野生型肾小球内的体内机械应力。通过拟议的研究，她将学习 器官芯片方法、质谱、超分辨率成像和活体显微镜。她有 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，将提供有关其作品转化价值的建议。冯博士将花费95% 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， 哈佛大学的研讨会和职业发展研讨会。拟议的项目将使冯博士 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。她的目标的推进将需要 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。