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.

项目摘要/摘要该K01赠款提案描述了一项为期五年的培训计划，旨在将Di Feng博士转变为成为独立的学术研究员。冯博士获得了博士学位在医学院威斯康星州在艾伦·考利（Allen Cowley）博士的心态下。她现在正在完成她的博士后研究金马丁·波拉克（Martin Pollak）博士的实验室，他是研究肾小球肾脏疾病遗传学的国际领导者。冯博士她的研究重点是阐明ACTN4突变的机制 - 这是一个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肾脏疾病形式（FSG）。无法更好地表征FSG构成的足细胞功能障碍已阻碍在建立更具体的个性化治疗方面的领域，超出了广泛的免疫抑制和反抗性高血压疗法。冯博士将她的研究重点放在突变的podocyte对机械应力是它在过滤肾小球中的血流时所经历的。她已经表明了由疾病引起的突变体给予的生物物理变化使足细胞变脆，表现出衰竭收缩力和肌动蛋白细胞骨架的破坏，以响应周期性拉伸。在当前的提议中 AIM 1试图进一步定义由突变体ACTN4引起的人类足细胞的响应受损伸展，但也要剪切应力。她将采用器官芯片方法更好地模拟这些压力同时量化了足细胞的相关生物力学和分子反应。 AIM 2将确定 ACTN4的翻译后磷酸化是否还会损害足细胞对机械的反应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。目标3计划使用CRISPR/CAS技术生成突变体ACTN4大鼠模型并使用插入显微镜测量突变体和WT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，质谱法，超分辨率成像和插入式显微镜。她有在Pollak博士的领导下，组建了一组导师和顾问团队，使领导者在这些相对学科中，包括Donald Ingber博士，Bruce Molitoris博士，Hanno Steen博士，Douglas Richardson博士以及博士罗杰·昂（Roger Tung）将提供与她作品的翻译价值有关的建议。冯博士将花费95％她在拟议的研究中获得的奖项，她的培训计划包括教学课程，下水道和哈佛的职业发展研讨会。拟议的项目将使冯博士为独立研究奖竞争，她计划将其调查结果从ACTN4和上述多学科方法，以进一步研究基于肌动蛋白的细胞骨架中的缺陷如何损害足细胞对体内机械压力的反应。她的目标的进步将采取位于哈佛大学的庞大资源和与思想领袖的联系，位于独特的环境中波士顿整合学术界和工业。