High-throughput palmitoyl-proteomics profiling of extracellular vesicles for identification of biomarkers for early detection of clinically significant prostate cancer

这项建议利用了跨机构合作和细胞外小泡的多学科专业知识。 (EV)生物学，以及在肿瘤学方面的成熟临床专业知识，以确定EV相关生物标记物 早期前列腺癌的检测、诊断及预后。肿瘤及其周围组织脱落的电动汽车 微环境是一种非常有希望的生物标志物来源，具有协助检测的潜力， 临床环境下癌症的诊断和预后。对于PC来说尤其如此，因为它有一个 迫切需要改进诊断，以减少不必要的活检和对惰性疾病的过度诊断 疾病，以及一种方法，将允许数千名诊断不足的男性在临床上 重要的PC接受适当的早期诊断。不幸的是，检测的严密性和重复性 肠道病毒相关生物标志物，具有适当的特异性和敏感性，以及一种检测方法 在北美的所有临床环境中提供可重复的结果，但尚未实现。我们最近 对大的囊泡小体(LO)的研究发现，以前的 未开发的EV相关生物标记物池，我们发现这些标记物预测了具有临床意义的PC。 作为一个在电动汽车生物学、个人电脑和生物标记物发现方面拥有交叉专业知识的团队，我们正在合作 寻求EV相关生物标志物，以足够的敏感性和特异性诊断PC 早期并区分惰性疾病和致命性疾病。我们的初步研究表明，肿瘤- 在PC患者中很容易检测到衍生的EVS。此外，对已知标记的评估表明 EV相关生物标志物在早期PC鉴别诊断中的巨大潜力。我们假设 将强大的EV相关标志物纳入现有的PC护理标准可以减少癌症- 致死性PC相关死亡早期改善避免惰性疾病过度治疗 这些患者类别的检测和鉴别诊断。 这个多学科团队将通过三个具体目标来解决这一假设：1)改进方法 从而实现了血浆和血清中LO的重复性分析。2)候选(棕榈酰)蛋白的发现 来自纯化的LO的生物标记物用于早期识别具有临床意义的PC和3)临床测试以评估 LO对早期前列腺癌的诊断和预后价值该项目下一步将包括几个 用于识别循环LO中的癌症特异性生物标记物并评估其 来自合作机构的PC患者在独立临床队列中的表现。我们的目标是 实施和验证一种严谨和可重复性的EV分析方法，具有很高的临床价值。

This proposal leverages trans-institutional collaboration and multidisciplinary expertise in extracellular vesicle (EV) biology, together with established clinical expertise in oncology, to identify EV-associated biomarkers for the detection, diagnosis and prognosis of early prostate cancer (PC). EVs shed by the tumor and its microenvironment are a very promising source of biomarkers, with the potential to assist with the detection, diagnosis and prognosis of cancer in the clinical setting. This is particularly true for PC where there is an immediate need for improved diagnostics to reduce unnecessary biopsies and over-diagnosis of indolent disease, and for a methodology that will allow thousands of under-diagnosed men who have clinically significant PC to receive appropriate early diagnosis. Unfortunately, the rigor and reproducibility of detecting EV-associated biomarkers with appropriate specificity and sensitivity, as well as a detection approach that provides reproducible results across all clinical settings in North America, have not been achieved. Our recent studies on large oncosomes (LO), which are atypically large EVs (1-10 m), have identified a previously unexplored pool of EV-associated biomarkers that we find predictive of clinically significant PC. As a team with cross-cutting expertise in EV biology, PC, and biomarker discovery, we are collaborating to pursue EV-associated biomarkers that can detect PC with sufficient sensitivity and specificity to diagnose PC early and to distinguish between indolent disease and lethal disease. Our preliminary studies reveal that tumor- derived EVs are readily detected in PC patients. Moreover, an assessment of known markers demonstrates enormous potential for EV-associated biomarkers in the differential diagnosis of early PC. We hypothesize that incorporating robust EV-associated markers into the existing standard of care for PC can reduce cancer- related deaths caused by lethal PC and avoid the overtreatment of indolent disease by improving early detection and differential diagnosis of these patient categories. This multidisciplinary team will address this hypothesis through three Specific Aims: 1) To refine methods leading to reproducible analysis of LO from plasma and serum. 2) Discovery of candidate (palmitoyl)protein biomarkers from purified LO for early identification of clinically significant PC and 3) Clinical testing to assess the diagnostic and prognostic value of LO in early prostate cancer. The project incorporates several next generation technologies and resources to identify cancer-specific biomarkers in circulating LO and assess their performance in independent clinical cohorts of PC patients from the collaborating institutions. Our objective is to implement and validate a rigorous and reproducible methodology for EV analysis with high clinical value.