Biological function of testican-2 in podocyte health

testican-2 在足细胞健康中的生物学功能

基本信息

批准号：
10599209
负责人：
Donghai Wen
金额：
$ 16.96万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
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/10599209
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

项目摘要

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.

项目摘要/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，我们最近将睾丸-2鉴定为肾脏释放到循环的足细胞衍生蛋白。此外，我们发现睾丸-2水平与估计的肾小球滤过率（EGFR）呈正相关。随着时间的推移，较高的睾丸基线水平与EGFR损失较少有关，从而推动了以下概念 Testican-2提供了有关肾脏健康和预后的见解。但是，睾丸-2在足细胞和肾脏未知。由Spock2基因编码，Testican-2是一种糖蛋白，是一种细胞外基质（ECM）的成分。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，adriamcyin诱导的足细胞损伤减弱。我们进一步找到睾丸-2结合了另一种ECM蛋白玻璃连素蛋白，并降低了其与整合素αVβ3的结合亲和力，A 足细胞表面上的粘附蛋白。将这些发现作为跳板，该建议将测试睾丸-2通过降低玻璃纤维蛋白介导的中心假设在足细胞健康中起功能作用整联蛋白αVβ3激活和信号传导：目标1）确定睾丸-2结合所需的蛋白质结构域玻璃纤维蛋白；目标2）确定睾丸-2对整联蛋白αVβ3激活和足细胞损伤的影响；目标3）评估糖尿病性肾病和原发性局灶性分段患者的睾丸-2表达肾小球硬化。候选人Wen博士深深致力于学术肾脏病的职业。他先前从事肾脏病学方面的广泛研究培训，特别关注肾脏生理学。自2018年追求肾脏学研究金以来，他将重点转移到了新领域 - 生物学功能睾丸-2在足细胞健康中，具有基本和翻译的方法。温博士的立即职业目标包括获得内容知识和在新兴技术领域的实践经验足细胞生物学；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，赠款和手稿写作的培训。温博士的长期职业目标是成为一个独立的身体科学家，他既采用基本和翻译的方法来发展针对足细胞损伤的患者和肾脏疾病的新治疗策略。他将从肾脏专家Eugene Rhee（导师； MGH Nephrology）的互补精神训练代谢和生物标志物；和马丁·普尔拉克（Martin Pollak 足细胞生物学。博士。 Wen，Rhee和Pollak将经常开会讨论科学和职业发展并清楚地确定了提出的研究的各个方面，这些方面将构成他的基础独立职业。咨询委员会将提供其他指导，评估进度并帮助计划未来的方向。