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: 10457294
• 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
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10457294
• 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; 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; 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; induced pluripotent stem cell; 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.

项目摘要/摘要 这份K01拨款建议书描述了一项为期五年的导师培训计划，旨在将狄峰博士转变为 成为一名独立的学术调查员。冯医生在重庆大学医学院获得博士学位。 在艾伦·考利博士的指导下，威斯康星州。她现在正在完成她的博士后研究项目 马丁·波拉克博士的实验室，他是研究肾小球疾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--一种重要的 细胞骨架蛋白--导致一种称为局灶性节段性肾小球硬化的肾小球疾病 (FSGS)。无法更好地描述FSGS背后的足细胞功能障碍阻碍了 在建立更具体、个性化的治疗方法方面，超越了广泛的免疫抑制和抗- 高血压治疗。冯博士将她的研究重点放在突变足细胞对 在过滤肾小球中的血液流动时，它会受到机械压力。到目前为止，她已经证明了 致病突变体ACTN4引起的生物物理变化使足细胞变得脆弱，表现出故障 收缩力量和肌动蛋白细胞骨架的破坏对周期性拉伸的反应。在目前的提案中， 目标1试图进一步定义由突变的ACTN4引起的人类足细胞的受损反应，而不仅仅是 拉伸的同时也要承受剪切应力。她将使用芯片上器官的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翻译后磷酸化是否也损害足细胞对机械刺激的反应 应激，使用小鼠模型和从这些小鼠分离的足细胞的生物力学研究。Aim 3计划 应用CRISPR/Cas技术建立ACTN4突变型大鼠模型，活体显微镜观察 突变型和WT肾小球内的活体机械应力。通过提议的研究，她将了解到 芯片上器官方法、质谱学、超分辨率成像和活体显微镜。她有 在波拉克博士的领导下，组建了一个由导师和顾问组成的团队，领导着这些学科的负责人， 包括唐纳德·英格伯博士、布鲁斯·莫利托里斯博士、汉诺·斯蒂恩博士、道格拉斯·理查森博士以及道格拉斯·理查森博士。 罗杰·董，她将就她的作品的翻译价值提供建议。冯博士将花费95%的时间 她在这一奖项下的时间用于拟议的研究，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 竞争独立研究奖，她计划将她在ACTN4和 以上多学科方法，以进一步研究基于肌动蛋白的细胞骨架缺陷如何损害 足细胞对体内经历的机械压力的反应。她的目标的实现将需要 在哈佛的巨大资源和与思想领袖的联系中，处于独特的环境中 整合了学术界和产业界的波士顿大学。