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

该提案利用了细胞外囊泡领域的跨机构合作和多学科专业知识， (EV)生物学，以及在肿瘤学方面建立的临床专业知识，以确定EV相关的生物标志物， 早期前列腺癌（PC）的检测、诊断和预后。由肿瘤脱落的EV及其 微环境是非常有前途的生物标志物来源，具有协助检测的潜力， 临床环境中癌症的诊断和预后。这对于PC来说尤其如此， 迫切需要改进诊断方法，以减少不必要的活检和对惰性肿瘤的过度诊断， 疾病，并为一种方法，将允许成千上万的诊断不足的男子谁有临床 重要PC接受适当的早期诊断。不幸的是，检测的严格性和可重复性 具有适当特异性和灵敏度的EV相关生物标志物，以及检测方法， 在北美的所有临床环境中提供可重复的结果，尚未实现。我们最近 对大癌小体（LO）的研究，即大体积EV（1-10 μ m），已经确定了以前的 未探索的EV相关生物标志物池，我们发现这些生物标志物可预测临床显著PC。 作为一个在EV生物学、PC和生物标志物发现方面拥有跨领域专业知识的团队，我们正在合作， 寻求EV相关的生物标志物，可以检测PC，具有足够的灵敏度和特异性来诊断PC 早期和区分惰性疾病和致命疾病。我们的初步研究显示肿瘤- 在PC患者中容易检测到衍生的EV。此外，对已知标志物的评估表明， 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- 相关死亡的致死性PC，并避免过度治疗惰性疾病，改善早期 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 导致来自血浆和血清的LO的可再现分析。2)发现候选（棕榈酰）蛋白 用于早期鉴定临床显著PC的来自纯化LO的生物标志物和3）临床测试以评估 LO对早期前列腺癌的诊断及预后价值。该项目包括几个未来 产生技术和资源，以确定循环LO中的癌症特异性生物标志物，并评估其 来自合作机构的PC患者的独立临床队列中的性能。我们的目标是 实施和验证具有高临床价值的EV分析的严格和可重复的方法。

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.