Dissecting the role of soluble a-Klotho in cardiovascular aging

项目概要/摘要 拟议的职业发展奖将促进和促进候选人的培训和发展， 独立调查员候选人：Kenneth Lim博士是一名临床翻译研究员， 肾脏科，马萨诸塞州总医院（MGH）和哈佛医学院（HMS）。拟定研究 整合了以患者为导向的研究，心肺运动测试（CPET）技术，蛋白质组学和 将计算生物学融入严格的训练计划中导师：Ravi Thadhani博士是 MGH的肾脏病学，HMS的医学教授和Partners的临床试验执行主任 健康护理他将与一个多学科专家小组一起担任主要导师 合作者研究：心脏病相关的心血管系统变化及其并发症是 慢性肾脏病（CKD）患者的主要死亡原因。尽管如此，目前还没有直接的 治疗这种疾病的方法。Klotho是一种存在于循环中的蛋白质， 多效衰老抑制作用。动物研究已经证明， Klotho可用于治疗CKD中的心血管疾病。然而，一些基本 在未来的人类干预研究之前，必须首先克服这些问题：首先，发表 迄今为止，研究循环Klotho与心血管结局的研究主要集中在 形态学改变，而明确的证据表明，衰老与减少 心血管储备功能第二，Klotho的各种循环同种型和 它们在心血管健康中的具体作用的性质仍不明确。此外，迄今为止的临床研究 评估循环Klotho的方法受到缺乏可靠Klotho测定的限制。建议的总体目标 因此，这项研究是为了弥合我们对循环Klotho在调节中的作用的理解的关键差距。 CKD患者心血管老化反应的研究。我们假设循环Klotho缺乏症是一种主要的 CKD患者心血管过早老化的决定因素。 在具体目标1中，我们将描述各种Klotho亚型与心血管疾病的关系。 结构和功能储备使用最先进的CPET技术。我们将定义循环水平 健康和晚期CKD（横断面）以及肾移植后（前瞻性）的Klotho亚型。 循环Klotho亚型水平将使用已建立的免疫沉淀法和Western印迹法进行评估。 印迹法此外，拟议的研究将提供一个强大的平台，以验证和确认新的 靶向蛋白质组学分析，我们已经开发了循环Klotho亚型的精密度评估。 在具体目标2中，我们将进行一项横断面研究，以描述循环Klotho 使用来自健康和CKD患者的人动脉进行过早的血管变化。以进一步确定 Klotho的治疗特性，我们将使用动脉外植体器官培养物进行干预性研究。

PROJECT SUMMARY/ABSTRACT The proposed career development award will foster and promote the candidate's training and evolution toward independent investigator. Candidate: Dr. Kenneth Lim is a clinical-translational investigator at the Division of Nephrology, Massachusetts General Hospital (MGH) and Harvard Medical School (HMS). The proposed study integrates patient-oriented research, cardiopulmonary exercise testing (CPET) technology, proteomics and computational biology embedded in a rigorous training plan. Mentorship: Dr. Ravi Thadhani is the Chief of Nephrology at the MGH, Professor of Medicine at HMS and Executive Director of Clinical Trials at Partners Healthcare. He will serve as the primary mentor in conjunction with a multidisciplinary team of expert collaborators. Research: Age-associated changes of the cardiovascular system and its complications are the leading cause of death in patients with chronic kidney disease (CKD). Despite this, there are currently no direct therapies available to treat this condition today. Klotho is a protein present in circulation that exerts highly pleiotropic aging suppressive effects. Animal studies have demonstrated promising therapeutic properties of Klotho that could be used for the treatment of cardiovascular disease in CKD. However, several fundamental problems must first be overcome before future human interventional studies can proceed: Firstly, published studies examining circulating Klotho with cardiovascular outcomes to-date have focused mainly on morphological alterations, while clear evidence has shown that aging is tightly associated with reduced cardiovascular functional reserve. Secondly, the precise levels of the various circulating isoforms of Klotho and the nature of their specific roles in cardiovascular health are still undefined. Additionally, clinical studies to-date assessing circulating Klotho are limited by the lack of a reliable Klotho assay. The overall aim of the proposed study is therefore to bridge a critical gap in our understanding of the role of circulating Klotho in the regulation of the cardiovascular aging response in CKD. We hypothesize that circulating Klotho deficiency is a major determinant of premature cardiovascular aging in CKD. In specific aim 1, we will characterize the relationship of the various Klotho isoforms with cardiovascular structure and functional reserve using state-of-the-art CPET technology. We will define levels of circulating Klotho isoforms in health and advanced CKD (cross-sectional), and after kidney transplantation (prospectively). Circulating Klotho isoform levels will be assessed using an established immunoprecipitation and western blotting method. Additionally, the proposed study will provide a robust platform to validate and confirm a new targeted proteomics assay that we have developed for the precision assessment of circulating Klotho isoforms. In specific aim 2, we will conduct a cross-section study to characterize the relationship of circulating Klotho with premature vascular changes using human arteries from healthy and CKD patients. To further determine therapeutic properties of Klotho, we will conduct an interventional study using arterial explant organ cultures.