BIN1 is a mediator and marker of cardiac reserve in heart failure.

项目摘要 超过500万美国人患有心力衰竭（HF），每年新增病例超过50万例 这是一种日益蔓延的流行病。然而，心力衰竭进展的病理生理学仍然知之甚少 因此，我们做出治疗决定的能力仍然有限。制定更具体和 需要与HF进展直接相关的预后生物标志物来改善临床管理。我有 最近发现，一个关键的支架蛋白，BIN 1，是重要的发病机制改变钙处理 在人类HF中。有了这些知识，我的长期目标是开发一种新的心脏储备的HF生物标志物 通过了解BIN 1的生物学和正常和患病心肌细胞中的钙处理。的 本申请的目的是了解BIN 1在调节细胞内钙瞬变中的作用， 心力衰竭，并使用BIN 1开发HF患者的诊断和预后测试。我的核心假设是 在衰竭的人类心肌细胞中，BIN 1的表达减少， 与心肌组织的恢复潜力相关。其理由是，成功完成 所提出的研究将填补基于BIN 1的HF钙瞬变调节知识的空白， 有助于开发HF患者心脏储备的创新诊断和预后测试。具体目标#1 目的是确定BIN 1在失败的人的Cav1.2运输和钙瞬时调节中的作用， 心肌细胞定量rtPCR、蛋白质印迹、ELISA、免疫荧光和T-小管分级分离将 用于评估失败和非失败人类中BIN 1和Cav1.2的细胞表达和定位。 心肌细胞在成年小鼠心肌细胞模型中，表面生物素化和活细胞钙成像将 用于研究BIN 1缺失心肌细胞中Cav1.2运输和钙瞬变。具体目标#2 确定心肌组织BIN 1表达作为终末期HF进展的诊断和预后检测 人类心肌病患者。定量免疫荧光将用于心脏活检，以确定 心室BIN 1表达水平与临床收缩参数和结果数据进行分析。具体 目的#3是鉴定血清BIN 1作为心肌病心脏储备的新的预后HF生物标志物 患者将通过心脏特异性ELISA在大规模心肌病人群中测量血清BIN 1 来自UCSF心力衰竭诊所的患者和年龄匹配的正常对照人群进行分析， 临床数据。预计这一贡献将确定BIN 1作为HF介导剂和生物标志物，以帮助跟踪HF的发生。 心力衰竭的进展和明显的临床结局。这一贡献将是巨大的，因为 这一发现将改变当前用于监测疾病进展心肌健康评估的范例 并指导心力衰竭的治疗策略。本申请中提出的研究具有创新性 因为它侧重于一种新的HF诊断和预后测试，组织和血清BIN 1水平， 心脏储备通过直接反映个体心肌细胞的生化健康。

Project Summary Over five million Americans have heart failure (HF), with more than 500,000 new cases added each year in what is a growing epidemic. Yet the pathophysiology of heart failure progression remains poorly understood and, as a result, our ability to make therapeutic decisions remains limited. Development of more specific and prognostic biomarkers that are directly tied to HF progression is required to improve clinical management. I have recently found that a key scaffolding protein, BIN1, is important to the pathogenesis of altered calcium handling in human HF. With this knowledge, my long term goal is to develop a novel HF biomarker of cardiac reserve through understanding the biology of BIN1 and calcium handling in normal and diseased cardiomyocytes. The objective of this particular application is to understand the role of BIN1 in regulation of the calcium transients in heart failure, and to use BIN1 to develop a diagnostic and prognostic test in HF patients. My central hypothesis is that BIN1 expression is reduced in failing human cardiomyocytes, and that BIN1 levels in tissue and serum correlate with recovery potential of myocardial tissue. The rationale is that successful completion of the proposed research will fill a gap in the knowledge of BIN1 based regulation of calcium transients in HF which helps to develop an innovative diagnostic and prognostic test of cardiac reserve in HF patients. Specific Aim #1 is to identify the role of BIN1 in Cav1.2 trafficking and calcium transient regulation in failing human cardiomyocytes. Quantitative rtPCR, western blot, ELISA, immunofluorescence, and T-tubule fractionation will be used to assess the cellular expression and localization of BIN1 and Cav1.2 in failing and non-failing human cardiomyocytes. In adult mouse cardiomyocyte models, surface biotinylation and live-cell calcium imaging will be used to study Cav1.2 trafficking and calcium transients in BIN1 depleted cardiomyocytes. Specific Aim #2 is to identify myocardial tissue BIN1 expression as a diagnostic and prognostic test of HF progression in end-stage human cardiomyopathy patients. Quantitative immunofluorescence will be used on heart biopsies to determine ventricular BIN1 expression level for analysis with clinical contractile parameters and outcome data. Specific Aim #3 is to identify serum BIN1 as a novel prognostic HF biomarker of cardiac reserve in cardiomyopathy patients. Serum BIN1 will be measured by cardiac specific ELISA in a large population of cardiomyopathy patients from the UCSF heart failure clinic and an age-matched normal control population for analysis with clinical data. The contribution is expected to identify BIN1 as a HF mediator and biomarker to help track the progression of heart failure and prognosticate clinical outcomes. This contribution will be significant because such discovery will shift current paradigm in myocardial health assessment for monitoring disease progression and guiding therapeutic strategies in heart failure. The research proposed in this application is innovative because it focuses on a new diagnostic and prognostic test of HF, tissue and serum BIN1 level, which assess cardiac reserve through directly reflecting the biochemical health of individual cardiomyocytes.