Background: Recent advancements in understanding plasma extracellular vesicles (EVs) and their role in disease biology have provided additional unique insights into the study of Colorectal Cancer (CRC). Methods: This study aimed to gain biological insights into disease progression from plasma-derived extracellular vesicle proteomic profiles of 80 patients (20 from each CRC stage I–IV) against 20 healthy age- and sex-matched controls using a high-resolution SWATH-MS proteomics with a reproducible centrifugation method to isolate plasma EVs. Results: We applied the High-Stringency Human Proteome Project (HPP) guidelines for SWATH-MS analysis, which refined our initial EV protein identification from 1362 proteins (10,993 peptides) to a more reliable and confident subset of 853 proteins (6231 peptides). In early-stage CRC, we identified 11 plasma EV proteins with differential expression between patients and healthy controls (three up-regulated and eight down-regulated), many of which are involved in key cancer hallmarks. Additionally, within the same cohort, we analysed EV proteins associated with tumour recurrence to identify potential prognostic indicators for CRC. A subset of up-regulated proteins associated with extracellular vesicle formation (GDI1, NSF, and TMED9) and the down-regulation of TSG101 suggest that micro-metastasis may have occurred earlier than previously anticipated. Discussion: By employing stringent proteomic analysis and a robust SWATH-MS approach, we identified dysregulated EV proteins that potentially indicate early-stage CRC and predict recurrence risk, including proteins involved in metabolism, cytoskeletal remodelling, and immune response. While our findings underline discrepancies with other studies due to differing isolation and stringency parameters, they provide valuable insights into the complexity of the EV proteome, emphasising the need for standardised protocols and larger, well-controlled studies to validate potential biomarkers.
背景：近期对血浆细胞外囊泡（EVs）及其在疾病生物学中作用的理解进展，为结直肠癌（CRC）研究提供了新的独特视角。方法：本研究旨在通过高分辨率SWATH-MS蛋白质组学技术，结合可重复的离心法分离血浆EVs，对80例患者（CRC I–IV期各20例）与20例年龄和性别匹配的健康对照的血浆来源细胞外囊泡蛋白质组谱进行分析，以获取疾病进展的生物学见解。结果：我们应用人类蛋白质组计划（HPP）的高严谨性指南进行SWATH-MS分析，将初始鉴定的1362种EV蛋白（10,993条肽段）精炼为更可靠、置信度更高的853种蛋白（6231条肽段）。在早期CRC中，我们鉴定出11种在患者与健康对照间差异表达的血浆EV蛋白（3种上调，8种下调），其中许多涉及关键癌症特征。此外，在同一队列中，我们分析了与肿瘤复发相关的EV蛋白，以识别CRC的潜在预后指标。与细胞外囊泡形成相关的上调蛋白（GDI1、NSF和TMED9）以及TSG101的下调提示，微转移可能比先前预期的更早发生。讨论：通过采用严格的蛋白质组学分析和稳健的SWATH-MS方法，我们鉴定出可能指示早期CRC并预测复发风险的失调EV蛋白，包括参与代谢、细胞骨架重塑和免疫反应的蛋白。尽管我们的研究结果因分离方法和严谨性参数的不同而与其他研究存在差异，但它们为EV蛋白质组的复杂性提供了有价值的见解，强调需要标准化方案以及更大规模、严格对照的研究来验证潜在的生物标志物。
肿瘤(癌症)患者之家