Calcification Propensity, using Dynamic Light Scattering, to Study Vascular Calcification in Patients with Advanced Chronic Kidney Disease

项目摘要/摘要 这项建议的目的是促进魏晨，医学博士，硕士作为一个独立的 具有研究慢性肾脏患者血管钙化(VC)专业知识的翻译研究人员 疾病(CKD)。CKD患者有较高的VC患病率，这是导致他们心血管疾病的原因之一 发病率和死亡率。在一定程度上，由于缺乏有效的VC生物标记物，目前还没有有效的 治疗以减缓其进展。最近，一种血清检测方法被用来测量钙化倾向， 可作为VC的生物标志物，指导新的治疗方法的开发。然而，它是未知的 钙化倾向和VC之间是否有直接关系，如本检测方法所测量的。Dr。 陈将通过研究钙化倾向与VC严重程度的关系来解决这个问题 以及导致VC的细胞机制。在与生物化学家本杰明·米勒博士(顾问)的合作下， 陈开发了一种新的、基于微孔板的分析方法，它使用动态光散射。他们的初步数据 证明了新的检测方法对VC的预测能力可能比旧的检测方法更强 使用散射比浊法。使用改进后的化验方法，陈博士将检验以下两个假设：1)血液透析 钙化倾向较高的患者冠状动脉钙化程度更大，进展更快 与倾向较低的人相比，动脉僵硬和更高的全因和心血管死亡率； 2)更高的钙化倾向与相关因子的更高的动脉RNA表达有关 在血管平滑肌细胞的骨软骨形成中，包括矮小相关转录因子-2(Runx2)和 骨钙素。第一个假设将在417名血液透析患者中进行验证，这些预测因素包括 终末期肾病的心律失常和心血管风险研究。为了检验第二个假设，Dr。 陈将招募100名正在接受动静脉通路创建手术的CKD患者，测量 血清钙化倾向，并在术中获取动脉样本(每个患者每次活检3-5 mm)以 检测动脉Runx2和骨钙素的RNA表达。这项提议将作出重大贡献。 旨在验证一种新的VC生物标记物，为VC的病理生理学提供新的见解，并具有 有可能指导新的治疗策略，以降低慢性肾脏病患者的死亡率。它的创新之处在于 陈博士和她的团队采用了一种新颖的分析方法和翻译方法来研究风险投资。此外，这一点 该计划将为陈博士提供机会来实现她的培训目标，即获得 从事VC研究的知识和技能，以及实施良好的临床实验室操作规范 作为纵向数据分析和临床研究设计。陈博士与她一起制定了这些培训目标 多学科指导团队，而实现这些目标对陈博士实现她的目标至关重要 长远的职业目标。这项K23奖励将支持拟议的培训和职业发展活动， 并允许陈博士成为独立的翻译研究员。

PROJECT SUMMARY/ABSTRACT The purpose of this proposal is to foster the development of Wei Chen, MD, MS as an independent translational researcher with the expertise to study vascular calcification (VC) in patients with chronic kidney disease (CKD). CKD patients have a high prevalence of VC, which contributes to their high cardiovascular morbidity and mortality. In part, due to the absence of a valid biomarker for VC, there is currently no effective treatment to slow its progression. Recently, a serum assay was developed to measure calcification propensity, and it may serve as a biomarker for VC to guide development of new therapies. However, it is unknown whether there is a direct relationship between calcification propensity, as measured by this assay, and VC. Dr. Chen will address this question by examining the association of calcification propensity with the severity of VC and the cellular mechanism leading to VC. In conjunction with a biochemist—Dr. Benjamin Miller (advisor), Dr. Chen developed a new, microplate-based assay that uses dynamic light scattering. Their preliminary data demonstrated that the new assay may have a greater predictive power for VC compared to the old assay that used nephelometry. Using the improved assay, Dr. Chen will test the following 2 hypotheses: 1) hemodialysis patients with higher calcification propensity have greater coronary arterial calcification, a faster progression of arterial stiffness and higher all-cause and cardiovascular mortality compared to those with lower propensity; and 2) higher calcification propensity is associated with higher arterial RNA expression of the factors involved in vascular smooth muscle cell osteochondrogenesis, including runt-related transcription factor-2 (runx2) and osteocalcin. The first hypothesis will be tested in 417 patients on hemodialysis from the Predictors of Arrhythmic and Cardiovascular Risk in End Stage Renal Disease study. To test the second hypothesis, Dr. Chen will recruit 100 CKD patients that are undergoing arteriovenous access creation surgeries, measure serum calcification propensity, and obtain intraoperative arterial samples (3-5mm per biopsy per patient) to measure arterial RNA expression of runx2 and osteocalcin. This proposal will make a significant contribution towards validating a novel VC biomarker, provide new insights into the pathophysiology of VC and has the potential to guide new treatment strategies to reduce mortality in patients with CKD. It is innovative because Dr. Chen and her team employ a novel assay and a translational approach to study VC. In addition, this proposal will provide opportunities for Dr. Chen to achieve her training objectives, which are to acquire knowledge and skills in conducting VC research, implementation of Good Clinical Laboratory Practice as well as longitudinal data-analyses and clinical study design. Dr. Chen developed these training objectives with her multidisciplinary mentoring team, and accomplishing these objectives is essential for Dr. Chen to achieve her long-term career goal. This K23 award will support the proposed training and career development activities, and allow Dr. Chen to become an independent translational researcher.