Biological function of testican-2 in podocyte health

testican-2 在足细胞健康中的生物学功能

• 批准号: 10867983
• 负责人: Donghai Wen
• 金额: $ 7.56万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10867983
• 关键词: Actins; Adhesions; Adriamycin PFS; Advisory Committees; Affinity; Albuminuria; Aorta; Area; Attenuated; Binding; Biological Markers; Biological Process; Biology; Blood; Boston; Cell surface; Cells; Chronic Kidney Failure; Circulation; Clinical Research; Creatinine; Cytoskeleton; Data; Dedications; Diabetic Nephropathy; Emerging Technologies; Enzyme-Linked Immunosorbent Assay; Etiology; Extracellular Matrix; Extracellular Matrix Proteins; Fellowship; Focal and Segmental Glomerulosclerosis; Future; General Population; Genes; Genetic; Glomerular Filtration Rate; Glycoproteins; Goals; Grant; Health; Human; ITGB3 gene; Immunoassay; Immunofluorescence Immunologic; In Vitro; Individual; Injury; Integrin alphaVbeta3; Interferometry; Interstitial Nephritis; Kidney; Kidney Diseases; Knowledge; Ligands; Manuscripts; Mass Spectrum Analysis; Measures; Mediating; Mentors; Mentorship; Microscopy; Mus; Nephrology; Patients; Peptides; Physicians; Physiology; Play; Prognosis; Proteins; Proteomics; Public Speaking; Publications; RGD (sequence); Renal glomerular disease; Research; Research Personnel; Research Training; Role; Sampling; Science; Scientist; Serum; Signal Transduction; Site-Directed Mutagenesis; Structure; Structure of renal vein; Surface; Tertiary Protein Structure; Testing; Therapeutic; Time; Tissue Sample; Tissues; Training; Vitronectin; Writing; aptamer; career; career development; cohort; experience; glomerulosclerosis; human subject; in vivo; inhibitor; insight; kidney biopsy; kidney metabolism; knock-down; multidisciplinary; novel; novel therapeutic intervention; overexpression; podocyte; population based; prognostic; protective effect; protein protein interaction; single-cell RNA sequencing; therapeutic target; tissue biomarkers; training opportunity; translational approach

PROJECT SUMMARY/ABSTRACT Using a proteomic screen of samples obtained from the aorta and renal vein of human subjects, we recently identified testican-2 as a podocyte-derived protein that is released by the kidney into circulation. Further, we found that testican-2 levels are positively correlated with estimated glomerular filtration rate (eGFR) and higher baseline levels of testican-2 are associated with less eGFR loss over time, advancing the concept that testican-2 provides insight on kidney health and prognosis. However, the biological function of testican-2 in podocytes and the kidney is unknown. Encoded by the SPOCK2 gene, testican-2 is a glycoprotein that is a component of the extracellular matrix (ECM). In preliminary data generated by the PI, we find that testican-2 attenuates podocyte injury induced by Adriamcyin both in cultured human podocytes and mice. We further find that testican-2 binds to another ECM protein vitronectin and reduces its binding affinity to integrin αVβ3, an adhesion protein on the podocyte surface. Using these findings as a springboard, this proposal will test the central hypothesis that testican-2 plays a functional role in podocyte health by reducing vitronectin-mediated integrin αVβ3 activation and signaling: Aim 1) To determine the protein domain required for testican-2 binding with vitronectin; Aim 2) To determine the impact of testican-2 on integrin αVβ3 activation and podocyte injury; and Aim 3) To assess testican-2 expression in patients with diabetic nephropathy and primary focal segmental glomerulosclerosis. The candidate, Dr. Wen, is deeply committed to a career in academic nephrology. He previously pursued broad-based research training in nephrology, with a particular focus on renal physiology. Since pursuing a nephrology fellowship in 2018, he has shifted his focus to a new area—the biological function of testican-2 in podocyte health, with both basic and translational approaches. Dr. Wen’s immediate career goals include acquiring content knowledge and hands-on experience with emerging technologies in the areas of podocyte biology; protein-protein interaction and structure; and blood and tissue biomarkers. In addition, he seeks training with public speaking and grant and manuscript writing. Dr. Wen’s long-term career goal is to become an independent physician-scientist who pursues both basic and translational approaches to develop new therapeutic strategies for patients with podocytes injury, and kidney disease in general. He will benefit from complementary mentorship from Eugene Rhee (mentor; Chief, MGH Nephrology), an expert in kidney metabolism and biomarkers; and Martin Pollak (co-mentor, Chief, BIDMC Nephrology), a leader in genetics and podocyte biology. Drs. Wen, Rhee, and Pollak will meet frequently to discuss both science and career development and have clearly identified aspects of the research proposed that will form the basis of his independent career. An advisory committee will provide additional guidance, evaluate progress, and help plan future directions.

项目总结/摘要 使用从人类受试者的主动脉和肾静脉获得的样品的蛋白质组学筛选， 最近鉴定出testican-2是一种足细胞衍生的蛋白质，由肾脏释放到循环中。此外，本发明还 我们发现睾丸素-2水平与估计的肾小球滤过率（eGFR）呈正相关， 睾丸素-2的基线水平越高，随着时间的推移eGFR损失越少，这一概念进一步发展， testican-2提供了对肾脏健康和预后的见解。然而，睾丸素-2的生物学功能， 足细胞和肾脏尚不清楚。由SPOCK 2基因编码，睾丸素-2是一种糖蛋白， 细胞外基质（ECM）的组成部分。在PI生成的初步数据中，我们发现testican-2 在培养的人足细胞和小鼠中减弱阿霉素诱导的足细胞损伤。我们进一步发现， 睾丸素-2与另一种ECM蛋白玻连蛋白结合，并降低其与整合素αVβ3的结合亲和力， 足细胞表面的粘附蛋白。以这些发现为出发点，本提案将测试 中心假设睾丸素-2通过减少玻连蛋白介导的 整合素αVβ3的激活和信号传导：目的1）确定睾丸素2结合所需的蛋白结构域 目的2）确定睾丸素-2对整合素αVβ3激活和足细胞损伤的影响; 目的3）探讨睾丸素-2（testican-2）在糖尿病肾病（DN）患者肾组织中的表达及意义 肾小球硬化候选人温博士致力于学术肾病学的事业。他 以前在肾脏学方面进行了广泛的研究培训，特别关注肾脏生理学。 自2018年获得肾脏病学奖学金以来，他已将重点转移到一个新的领域-生物功能 testican-2在足细胞健康中的作用，包括基本方法和翻译方法。温博士的近期职业目标 包括获取内容知识和实践经验，与新兴技术领域， 足细胞生物学;蛋白质-蛋白质相互作用和结构;以及血液和组织生物标志物。此外他 寻求公开演讲、赠款和手稿写作方面的培训。温博士的长期职业目标是 成为一个独立的物理学家，科学家谁追求的基本和转化的方法来发展 足细胞损伤和一般肾脏疾病患者的新治疗策略。他将受益于 来自肾脏专家尤金·李（导师; MGH肾脏科主任）的补充指导 和Martin Pollak（共同导师，主任，BIDMC肾病），遗传学和生物标志物领域的领导者， 足细胞生物学文博士、李承晚博士和波拉克博士将经常会面，讨论科学和职业问题 发展，并已明确确定的研究方面提出，将形成他的基础， 独立的职业生涯。咨询委员会将提供额外指导、评估进展并帮助计划 未来的方向