Proteome and Transcriptome Markers of Hypertension in Urine and Plasma Exosomes

描述（由申请人提供）：生物标志物发现带来个性化医疗的前景，即改善诊断和预后，已经被广泛传播，并且最多取得了轻微的成功。由于血液样本的可及性和高灵敏度蛋白质组学技术的快速发展，用于生物标志物发现的最常用工具是血浆蛋白质组学。在血浆样品中发现生物标志物的主要障碍是众所周知的动态范围问题，其中高丰度蛋白质（如血清白蛋白）的丰度是最不浓缩的（可能是最重要的）标记蛋白的1010倍。提出的研究计划概述了一种根本不同的新颖策略，该策略结合了转录组学和蛋白质组学方法，旨在规避血浆蛋白质组学的动态范围问题。该研究的具体目标是开发一种结合转录组学和蛋白质组学的方法来测量分离的血浆微囊泡和尿外泌体中的疾病特异性RNA和蛋白质特征。这些脂质颗粒含有蛋白质和RNA，已知在癌症和炎症性疾病中分泌率较高；因此，它们目前正在作为生物标志物载体进行研究。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特征提供了强有力的协同方法。为了进一步提高分析特异性，将从血浆和尿液样本中分离出细胞和组织特异性微粒，以获得诊断标志物、机制信息和监测治疗反应的标志物。最后，生物标志物将在高血压中被识别和验证，高血压是一种常见的人类疾病，如果不治疗，它会导致中风、心力衰竭和肾衰竭的巨大负担。高血压受试者将接受控制饮食以进行盐敏感性分类，并收集血浆和尿液进行随后的分子分析和统计相关性。预计将发现一组新的生物标志物（包括蛋白质和RNA），并将其作为诊断、预后和治疗反应的标志物，从而改善高血压的治疗。

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.