microRNAs in Prostate-derived Exosomes as Prognostic Biomarkers and Mediators of Intercellular Communication

项目总结/摘要 与其他癌症相比，前列腺癌（PCa）的治疗由于其高风险而造成独特的问题。 无痛性疾病的流行率和高治疗发病率。PCa是最常见的癌症诊断 也是美国男性癌症相关死亡的第二大原因，但42%的50岁或以上的美国男性 在尸检研究中发现死于其他原因的人有PCa。因此，临床医生必须 准确地确定哪些患者将进展为致命的转移性PCa，哪些不会。明确 诊断和危险分层是基于多芯活检结果。然而，PCa通常是多灶性的， 在某些情况下，活检可能会错过异质性肿瘤，并且活检可能会错过包含侵袭性肿瘤的区域。这种不确定性使得 低风险前列腺癌的过度治疗因此，临床上迫切需要准确的预干预 PCa预后测试，使临床医生可以治疗高风险患者，但从明确的低风险患者 治疗发病率。为了满足这一临床需求，本提案的第一个目标是发现和 表征前列腺来源的预后血清外泌体微小RNA（miR）。外泌体是脂质双层 从细胞分泌并在细胞间通讯中起作用的囊泡，但已被用于其 潜在的预后价值。这些实验利用了为治疗前收集的PCa血清群组。 以前资助的研究。我们的目标是利用外来体开发一种敏感和特异的预后测试， PCa。先前发表的数据表明前列腺miR的预后作用，其选择性地加载到 因此，在循环外泌体中可能存在对PCa特异性的miR群体。 除了外泌体miR的预后潜力之外，这些囊泡在细胞间相互作用中具有生物学功能。 细胞通讯此外，外泌体已经涉及在肿瘤中的肿瘤进展和转移。 多种癌症，包括前列腺癌。最常见的PCa形式来自于前列腺的上皮细胞（PrE）， 腺泡，被纤维肌基质包围。间质的功能是支持上皮细胞 在正常前列腺中促进生长和分化，但在PCa中促进肿瘤。这些促进肿瘤的 基质细胞被称为癌症相关成纤维细胞（CAF）。本建议的第二个目标是 阐明前列腺上皮细胞CAF释放的exosomes中microRNA的生物学作用 侵略性人前列腺癌根治术后原代培养的前列腺上皮细胞和基质细胞 手术组织将被用作模型系统以确定上皮细胞生长，增殖， 和用来自CAF的外来体处理后的细胞因子产生。从细胞中释放的外泌体中的miR CAF将被敲低或过表达以确定外泌体对PrE细胞的作用机制。 该项目使用患者衍生的模型，以翻译和多学科的方法开发一种非 侵袭性血清外泌体miR检测用于PCa预后并确定miR的肿瘤促进作用 从CAF释放。

PROJECT SUMMARY/ABSTRACT Compared to other cancers, the treatment of prostate cancer (PCa) poses a unique problem due to its high prevalence of indolent disease and high rates of treatment morbidity. PCa is the most common cancer diagnosis and the second leading cause of cancer-related mortality among US men, but 42% of US men age 50 or older that died from other causes were found to have PCa in autopsy studies. Therefore, it is essential for clinicians to accurately determine which patients will progress to lethal, metastatic PCa and which will not. Definitive diagnosis and risk stratification is based off of multi-core biopsy results. However, PCa is often multifocal with heterogeneous tumors, and the biopsy may miss areas that contain aggressive tumor. This uncertainty has led to the over-treatment of low-risk PCa. Therefore, there is an urgent clinical need for accurate pre-intervention PCa prognostic testing so that clinicians can treat high-risk patients but spare low-risk patients from the definitive treatment morbidities. To address this clinical need, the first objective of this proposal is to discover and characterize prognostic serum exosomal microRNAs (miRs) of prostatic origin. Exosomes are lipid-bilayer vesicles that are secreted from cells and function in intercellular communication but have been exploited for their potential prognostic value. These experiments capitalize on a pretreatment PCa sera cohort collected for a previously funded study. Our goal is to utilize exosomes to develop a sensitive and specific prognostic test for PCa. Previously published data suggests a prognostic role for prostate miRs, which are selectively loaded into these vesicles, so there may be a population of miRs in circulating exosomes that are specific to PCa. In addition to the prognostic potential of exosomal miRs, these vesicles have biological functions in cell-to- cell communication. Furthermore, exosomes have been implicated in tumor progression and metastasis in multiple cancers, including PCa. The most common form of PCa arises from the epithelial cells (PrE) of the glandular acini, which are surrounded by a fibromuscular stroma. The stroma functions to support epithelial growth and differentiation in the normal prostate but becomes tumor-promoting in PCa. These tumor-promoting stromal cells are termed cancer-associated fibroblasts (CAF). The second objective of this proposal is to elucidate the biological role of microRNAs in exosomes released from CAF on prostate epithelial cell aggressiveness. Human primary prostate epithelial and stromal cells derived from radical prostatectomy surgery tissue will be used as a model system to determine the changes in epithelial cell growth, proliferation, and cytokine production after treatment with exosomes from CAF. The miRs within the exosomes released from CAF will be knocked down or overexpressed to determine the mechanism of exosome action on PrE cells. This project uses patient-derived models in a translational and multi-disciplinary approach to develop a non- invasive serum exosome miR test for PCa prognosis and to determine the tumor-promoting effects of miRs released from CAF.