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

描述（由申请人提供）：超过500万美国人患有心力衰竭（HF），每年新增50多万例，这是一种日益增长的流行病。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。需要开发与HF进展直接相关的更特异性和预后生物标志物以改善临床管理。我最近发现，一个关键的支架蛋白，BIN 1，是重要的改变人类HF钙处理的发病机制。有了这些知识，我的长期目标是通过了解BIN 1的生物学和正常和患病心肌细胞中的钙处理来开发一种新的心脏储备的HF生物标志物。该特定应用的目的是理解BIN 1在细胞内钙瞬变调节中的作用。 心力衰竭，并使用BIN 1开发HF患者的诊断和预后测试。我的中心假设是，BIN 1的表达在失败的人心肌细胞中减少，并且组织和血清中的BIN 1水平与心肌组织的恢复潜力相关。其基本原理是，成功完成拟议的研究将填补基于BIN 1的HF钙瞬变调节知识的空白，这有助于开发HF患者心脏储备的创新诊断和预后测试。具体目标#1是鉴定BIN 1在衰竭的人心肌细胞中Cav1.2运输和钙瞬时调节中的作用。将使用定量rtPCR、蛋白质印迹、ELISA、免疫荧光和T-小管分级来评估BIN 1和Cav1.2在衰竭和非衰竭人心肌细胞中的细胞表达和定位。在成年小鼠心肌细胞模型中，将使用表面生物素化和活细胞钙成像来研究BIN 1耗尽的心肌细胞中的Cav1.2运输和钙瞬变。具体目标#2是鉴定心肌组织BIN 1表达作为终末期人类心肌病患者中HF进展的诊断和预后测试。定量免疫荧光将用于心脏活检，以确定心室BIN 1表达水平，用于分析临床收缩参数和结局数据。具体目标#3是鉴定血清BIN 1作为心肌病患者心脏储备的新型预后HF生物标志物。将通过心脏特异性ELISA在来自UCSF心力衰竭诊所的大量心肌病患者人群和年龄匹配的正常对照人群中测量血清BIN 1，以分析临床数据。预计这一贡献将确定BIN 1作为HF介体和生物标志物，以帮助跟踪心力衰竭的进展和明确的临床结局。这一贡献将是重要的，因为这一发现将改变当前心肌健康评估的范式，以监测疾病进展并指导心力衰竭的治疗策略。该申请中提出的研究是创新的，因为它专注于HF，组织和血清BIN 1水平的新诊断和预后测试，通过直接反映个体心肌细胞的生化健康来评估心脏储备。

DESCRIPTION (provided by applicant): 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.