Proteome and Transcriptome Markers of Hypertension in Urine and Plasma Exosomes

描述(由申请人提供)：生物标记物发现导致个性化医学的前景，即改善诊断和预后，已得到广泛传播，充其量只取得了轻微的成功。由于血液样本的可获得性和高度敏感的蛋白质组学技术的快速发展，最常用的生物标记物发现工具是血浆蛋白质组学。在血浆样本中发现生物标记物的主要障碍是众所周知的动态范围问题，其中高度丰富的蛋白质，如血清白蛋白，比浓度最低，可能是最重要的标记蛋白的丰度高1010倍。拟议的研究计划概述了一种根本不同的新策略，它结合了转录组和蛋白质组方法，旨在绕过血浆蛋白质组学中的动态范围问题。这项研究的具体目标是开发一种结合转录和蛋白质组学的方法来测量分离的血浆微泡和尿外切体中的疾病特异性RNA和蛋白质特征。这些脂质颗粒同时含有蛋白质和RNA，已知在癌症和炎症性疾病中分泌速度较高；因此，它们目前正被作为生物标志物载体进行研究。所提出的蛋白质和RNA分析的互补策略为阐明具有高特异性和敏感性的疾病特异性信号提供了一种强大的协同方法。为了进一步提高分析的特异性，将从血浆和尿样中分离出细胞和组织特异性微粒，以获取诊断标志物、机制信息和监测治疗反应的标志物。最后，生物标记物将在高血压中得到识别和验证，高血压是一种常见的人类疾病，除非接受治疗，否则会造成中风、心力衰竭和肾功能衰竭的巨大负担。高血压患者将接受控制饮食，以进行盐敏感性分类，并采集血浆和尿液，以便随后进行分子分析和统计关联。预计一组新的生物标记物(蛋白质和RNA)将被发现并被证实为诊断、预后和治疗反应的标记物，将使高血压的治疗得到改进。 公共卫生相关性：拟议研究的目标是开发一种新的方法来发现疾病的分子标记，以绕过现有的障碍。通过分析从血液和尿液中分离出的脂肪颗粒中的蛋白质和RNA，将发现新的疾病标志物，以改善诊断、预后和治疗反应的预测，即改善个性化药物。新的方法将被应用于揭示高血压患者盐敏感性和治疗反应的生物标记物。

DESCRIPTION (provided by applicant): The promise of biomarker discovery leading to personalized medicine, i.e. improved diagnosis and prognosis, has been widely disseminated and has met with mild success, at best. The most common tool used for biomarker discovery is plasma proteomics because of the accessibility of blood samples and the rapid development of highly sensitive proteomics technologies. The major obstacle for biomarker discovery in plasma samples is the well known dynamic range issue where highly abundant proteins such as serum albumin are 1010 times more abundant than the least concentrated, perhaps most important, marker proteins. The proposed research plan outlines a fundamentally different and novel strategy that combines both transcriptomic and proteomic methodologies that is designed to circumvent the dynamic range issue in plasma proteomics. The specific goal of the proposed research is to develop a combined transcriptomic and proteomics approach to measure disease specific RNA and protein signatures in isolated plasma microvesicles and urinary exosomes. These lipid particles contain both protein and RNA and are known to be secreted at higher rates in cancer and inflammatory diseases; hence, they are currently being investigated as biomarker vectors. The proposed complementary strategy of protein and RNA analysis provides a powerful synergistic approach for elucidating disease- specific signatures with high specificity and sensitivity. To further increase analytical specificity, cell- and tissue-specific microparticles will be isolated from plasma and urine samples to acquire diagnostic markers, mechanistic information, and markers to monitor therapeutic response. Finally, biomarkers will be identified and validated in hypertension, a common human condition, which contributes to the enormous burden of stroke, heart failure, and renal failure unless treated. Hypertensive subjects will be subjected to controlled diet for classification of salt-sensitivity and plasma and urine collection for subsequent molecular analysis and statistical correlation. It is anticipated that a panel of novel biomarkers (both protein and RNA) will be discovered and validated as markers for diagnostic, prognostic, and therapeutic response that will enable improved treatment of hypertension. PUBLIC HEALTH RELEVANCE: The goal of the proposed research is to develop a novel method for discovery of molecular markers of disease that circumvents existing obstacles. Through analysis of proteins and RNA found in lipid particles isolated from blood and urine, new markers of disease will be discovered that improve diagnosis, prognosis, and prediction of response to therapy; that is, improve personalized medicine. The new methodology will be applied to reveal biomarkers of salt- sensitivity and therapeutic response in hypertensive subjects.