Circulating Exosome microRNA as Markers of Severe Sarcoidosis Phenotypes

项目摘要 结节病是一种成人发病的全身性炎症性疾病，可损害重要器官的功能。 肺部是最常见的影响，表现是很容易发现与传统的诊断 测试（例如，胸部X线摄影）。然而，心脏结节病[心脏结节病（CS）]和中央 神经系统[神经结节病（NS）]存在于结节病患者的重要子集中，这些 症状更难检测。重要的是要确定CS和NS，因为这些参与 器官对结节病的总体发病率和死亡率有显著影响。我们的初步研究 实验室表明，循环亚细胞组分，称为外来体，含有小RNA （microRNA）可以作为结节病的生物标志物，并可能能够识别哪些器官是结节病。 影响。我们认为，外泌体衍生的microRNA将使我们能够特异性地检测CS和NS。目标1 我们建议使用从结节病患者获得的血浆样本， 参与确定循环内体衍生的microRNA是否可靠地检测CS，我们将进一步 确定CS患者中循环外泌体来源的microRNA相对于患有CS的患者的特异性， 缺血性心脏病该提案的目的2将确定循环外泌体衍生的microRNA是否可以被 作为NS的生物标志物，并将进一步确定这些生物标志物是否可以区分NS和CS。这些 研究将使用NIH BioLINCC储存库提供的库存样本进行， 从最近完成的NIH赞助的α-1抗胰蛋白酶基因组研究中获得的样本 缺乏症和结节病（GRADS）试验。对于每一项研究，目标外泌体来源的microRNA表达将 使用下一代测序技术和最有前途的 microRNA生物标志物将在第二个队列中使用定量实时聚合酶链进行验证 反应（qRT-PCR）。这些研究将指导后续前瞻性试验的设计， 循环外泌体衍生的microRNA作为器官特异性结节病生物标志物。

PROJECT SUMMARY Sarcoidosis is an adult onset systemic inflammatory disease that can compromise the function of vital organs. The lungs are most commonly affected, a manifestation that is readily detected with conventional diagnostic tests (e.g., chest radiography). However, sarcoidosis of the heart [cardiac sarcoidosis (CS)] and central nervous system [neurosarcoidosis (NS)] is present in a significant subset of sarcoidosis patients and these manifestations are more difficult to detect. It is important to identify CS and NS because involvement of these organs contributes significantly to the overall morbidity and mortality of sarcoidosis. Preliminary studies in our laboratory indicate that circulating subcellular components, referred to as exosomes, contain small RNA (microRNA) that can serve as biomarkers of sarcoidosis, and may be able to identify which organs are affected. We posit that exosome-derived microRNA will allow us to specifically detect CS and NS. In Aim 1 we propose to use plasma samples obtained from sarcoidosis patients with well-documented cardiac involvement to determine if circulating endosome-derived microRNA reliably detects CS, and we will further determine the specificity of circulating exosome-derived microRNA in CS patients relative to patients with ischemic heart disease. Aim 2 of the proposal will establish if circulating exosome-derived microRNA can be used as biomarkers of NS, and will further determine if these biomarkers can distinguish NS from CS. These studies will be performed using banked samples provided by the NIH BioLINCC repository, and banked samples available from the recently completed NIH sponsored Genomic Research of Alpha-1 Antitrypsin Deficiency and Sarcoidosis (GRADS) trial. For each research Aim exosome-derived microRNA expression will be assessed quantitatively in a smaller test cohort using next-generation sequencing and the most promising microRNA biomarkers will be validated in a second cohort using quantitative real-time polymerase chain reaction (qRT-PCR). These studies will guide the design of subsequent prospective trials that would establish circulating exosome-derived microRNA as organ-specific sarcoidosis biomarkers.