A Putative Heparin Receptor in Smooth Muscle Cells

描述(申请人提供)：在西方国家，心血管疾病是导致死亡人数最多的疾病。此外，2010年，血管疾病患者的治疗预计将占医疗支出的3000亿美元以上。有证据表明，药物肝素可以有效地短期治疗许多心血管疾病的症状，实验室研究表明，肝素减缓了血管平滑肌细胞的生长，血管平滑肌细胞是晚期血管疾病的组成部分。PI实验室的长期研究目标是了解肝素治疗如何导致血管细胞功能的变化。这些知识可能有助于开发血管疾病的先进治疗方法。此前，PI的实验室开发了模拟肝素对培养的血管细胞影响的抗体，这些抗体已被用于研究血管平滑肌细胞增殖如何受到抑制的证据。目前这项提案的研究目的是通过首先评估肝素的整个反应而不仅仅是预先预测的那些特定变化来增强这种理解。这将涉及使用阵列技术，该技术可以同时检查许多不同基因的变化。使用这项技术确定的具体目标将得到进一步评估。其次，将使用为蛋白质鉴定和序列测定开发的质谱学技术来鉴定肝素受体的蛋白质和基因。来自已鉴定蛋白质的序列信息将被用于创建新的工具，这些工具将被用于进一步确认肝素受体的鉴定。第三，基于我们实验室以前的工作，从肝素受体中发现了一个可能的信号系统，我们建议比较通过肝素受体和触发类似细胞内途径的受体发出信号的结果，以增加我们对肝素信号的理解。第四，心血管疾病有一个重要的炎症成分，这也可能是肝素作用的目标，这与我们的初步结果一致。将评估肝素治疗降低血管平滑肌细胞炎症反应的能力。此外，在同时使用和不使用肝素处理的培养内皮细胞中，将检测炎性介质诱导的特定基因表达的变化，这些变化涵盖了一系列显著改变的基因类型。如果肝素治疗导致基因表达的炎性变化减少，数据将支持这样的假设，即肝素信号在血管系统中与炎症信号相反。结合这项工作的其他结果，这些数据将提供对肝素改变血管细胞行为的机制的坚实理解，并为肝素受体成为血管疾病高级治疗的靶点提供建议。 公共卫生相关性：心血管疾病死亡和对患有这些疾病的个人的治疗支出是卫生保健成本的主要因素。肝素被认为是一种可以减缓疾病进展的治疗方法，但我们对肝素如何帮助减缓疾病进展的了解有限。这项拟议的研究旨在增加我们对药物肝素如何在分子水平上减轻血管炎症的理解，因此应该有助于开发新的血管疾病治疗策略。

DESCRIPTION (provided by applicant): Cardiovascular diseases account for more deaths than any other disease in western countries. In addition, treatment of individuals with vascular diseases is expected to account for more than $300 billion in health care spending in 2010. Evidence suggests that the drug heparin can be an effective short-term treatment for many of the symptoms of cardiovascular disease, and laboratory studies indicate that heparin slows vascular smooth muscle cell growth, a component of late stage vascular disease. The long-term research goals in the PI's laboratory are to understand how heparin treatment results in changes in vascular cell function. Such knowledge is likely to contribute to the development of advanced treatments for vascular diseases. Previously, the PI's laboratory developed antibodies that mimic heparin effects on vascular cells in culture and those antibodies have been used in studies that resulted in evidence for how vascular smooth muscle cell proliferation is inhibited. The research aims for the current proposal will enhance that understanding by first evaluating the entire response to heparin rather than only those specific changes that were predicted in advance. This will involve using array technology that can examine changes in many different genes at one time. Specific targets identified using this technology will be evaluated further. Second, the protein and gene for the heparin receptor will be identified using mass spectrometry techniques developed for protein identification and sequence determination. Sequence information from the identified protein will be used to create new tools that will be used to further confirm the identification of the heparin receptor. Third, based on previous work from our laboratory that identified a likely signal system from the heparin receptor, we propose to compare the results of signaling through the heparin receptor and receptors that trigger similar intracellular pathways to increase our understanding of heparin signaling. Fourth, cardiovascular disease has a significant inflammatory component that is likely to also be a target of heparin action, consistent with our preliminary results. The ability of heparin treatment to decrease inflammatory responses in vascular smooth muscle cells will be evaluated. In addition, changes in expression of specific genes induced by inflammatory mediators that cover a range of significantly altered gene types will be examined in cultured endothelial cells with and without concurrent heparin treatment. If heparin treatment results in decreased inflammatory changes in gene expression, the data will support the hypothesis that heparin signaling acts counter to inflammatory signaling in the vasculature. Coupled with the other results of this work, these data will provide a solid understanding of the mechanisms by which heparin alters vascular cell behavior and suggestions for ways in which the heparin receptor could be a target for advanced therapies for vascular diseases. PUBLIC HEALTH RELEVANCE: Deaths from cardiovascular disease and spending on treatments for individuals suffering from these diseases are major factors in the costs of health care. The drug heparin has been suggested as a treatment that could decrease progression of the disease, but we have a limited understanding of how heparin might work to help slow disease progress. The proposed research is designed to increase our understanding of how the drug heparin works at the molecular level to decrease inflammation in blood vessels, and should therefore facilitate development of new treatment strategies for vascular disease.