Dissecting the role of soluble a-Klotho in cardiovascular aging

项目摘要/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，临床翻译研究员 马萨诸塞州总医院(MGH)和哈佛医学院(HMS)的肾脏病。建议进行的研究 将以患者为中心的研究、心肺运动试验(CPET)技术、蛋白质组学和 在严格的培训计划中嵌入了计算生物学。导师：拉维·塔达尼博士是 MGH的肾病学，HMS的医学教授和Partners的临床试验执行董事 医疗保健。他将与一个由多学科专家组成的团队一起担任主要导师 合作者。研究：与年龄相关的心血管系统变化及其并发症是 慢性肾脏疾病(CKD)患者的主要死因。尽管如此，目前还没有直接的 目前可用于治疗这种疾病的治疗方法。Klotho是一种存在于循环中的蛋白质，它高度发挥作用 多效性抗衰老作用。动物研究已经证明了有希望的治疗特性 Klotho可用于治疗CKD的心血管疾病。然而，有几个基本问题 在未来的人类干预研究可以进行之前，必须首先解决问题：第一，发表 到目前为止，研究循环Klotho与心血管后果的研究主要集中在 形态改变，虽然明确的证据表明，衰老与减少密切相关 心血管功能储备。其次，Klotho和Klotho的各种循环亚型的精确水平 它们在心血管健康中的具体作用的性质仍然不确定。此外，迄今为止的临床研究 由于缺乏可靠的Klotho检测，对循环Klotho的评估受到限制。建议的总体目标是 因此，研究是为了弥合我们对循环Klotho在调节中的作用的理解中的一个关键差距 CKD患者的心血管衰老反应。我们假设循环中的Klotho缺乏症是一个主要的 慢性肾脏病心血管早衰的决定因素。 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 采用最先进的CPET技术进行结构和功能储备。我们将定义循环的级别 Klotho亚型在健康和晚期CKD中(横断面)，以及肾移植后(前瞻性)。 循环中的Klotho亚型水平将使用已建立的免疫沉淀和Western进行评估 吸墨法。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 我们为精确评估循环中的Klotho亚型而开发的靶向蛋白质组学分析。 在具体目标2中，我们将进行横断面研究，以表征循环Klotho的关系 使用健康和CKD患者的人体动脉进行早期血管变化的研究。以进一步确定 对于Klotho的治疗特性，我们将使用动脉外植体器官培养进行介入性研究。

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.