PO.CH02.01 · 化学
头颈部肿瘤来源的小细胞外囊泡的潜在临床应用价值
Potential clinical utility of small extracellular vesicles derived from head and neck tumors
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
小细胞外囊泡（EVs）携带具有选择性的蛋白质货物，能够反映其亲代肿瘤细胞的生理状态，使其成为头颈部癌（HNC）生物标志物的一个有前景的来源。已知HNC来源的小EVs参与多个关键病理过程，包括免疫抑制、治疗耐药和转移潜能。唾液和血浆作为易于获取且微创的生物体液，可作为肿瘤来源生物标志物的替代来源，以增强临床监测。然而，从这些体液中富集得到的小EVs能否准确代表肿瘤来源的EVs，目前尚不清楚。本研究优化了从HNC肿瘤组织中分离小EVs的方法，并评估了肿瘤来源、唾液来源和血浆来源EVs之间的蛋白质组学重叠情况。
采用超速离心法从肿瘤组织和唾液中分离小EVs，采用尺寸排阻色谱法从血浆中分离小EVs。通过冷冻透射电子显微镜（cryo-TEM）和纳米颗粒追踪分析（NTA）评估小EVs的形态、大小和浓度。采用SWATH质谱对小EVs的蛋白质组进行分析和定量。对重叠蛋白和特有蛋白进行功能分析，以确定其生物学相关性。
从肿瘤组织、唾液和血浆中分离得到的小EVs均呈现典型的脂质双分子层形态，大小范围为40至200 nm。所鉴定的蛋白质货物中，超过60%在全部三种来源之间共享。这些共享蛋白显著富集于与癌症进展相关的通路，包括mTORC1信号通路、凝血、补体、上皮-间质转化以及PI3K/AKT/mTOR信号通路。为支持蛋白质组学结果的可靠性，通过免疫印迹进一步验证了一部分蛋白。
肿瘤组织、唾液和血浆来源的小EVs之间存在大量蛋白质组学重叠，这支持了将体液EVs用作肿瘤EVs替代物的可行性。唾液和血浆易于获取，是用于生物标志物发现和临床监测的可行、非侵入性来源。这些发现为推进HNC中的液体活检方法奠定了坚实基础。
查看英文原文 English abstract
Small extracellular vesicles (EVs) carry selective protein cargo that reflect the physiological state of their parent tumor cells, making them a promising source of biomarkers for head and neck cancer (HNC). HNC-derived small EVs are known to contribute to key pathological processes, including immunosuppression, therapy resistance, and metastatic potential. Saliva and plasma, as accessible and minimally invasive biofluids, represent surrogates for tumor-derived biomarkers to enhance clinical monitoring. However, it remains unclear whether small EVs enriched from these fluids accurately represent tumor-derived EVs. This study optimised small EV isolation from HNC tumor tissue and evaluated proteomic overlap between tumor-, saliva-, and plasma-derived EVs.
Small EVs were isolated from tumor tissue and saliva by ultracentrifugation and from plasma using size exclusion chromatography. The morphology, size and concentration of small EVs were assessed by cryogenic Transmission electron microscopy (cryo-TEM) and nanoparticle tracking analysis (NTA). The proteome of small EVs was profiled and quantified using SWATH mass spectrometry. Functional analysis of overlapping and unique proteins was performed to determine their biological relevance.
Small EVs isolated from tumor tissue, saliva and plasma exhibited typical lipid bilayer morphology with sizes ranging from 40 to 200 nm. More than 60% of the identified protein cargoes were shared across all three sources. These shared proteins were significantly enriched in pathways associated with cancer progression, including mTORC1 signalling, coagulation, complement, epithelial-mesenchymal transition, and PI3K/AKT/mTOR signalling. To support the reliability of the proteomic results, a subset of proteins was further validated by immunoblotting.
The substantial proteomic overlap between small EVs from tumor tissue, saliva, and plasma supports the feasibility of using biofluid EVs as surrogates for tumor EVs. Saliva and plasma, being easily accessible, represent viable, non-invasive sources for biomarker discovery and clinical monitoring. These findings provide a strong foundation for advancing liquid biopsy approaches in HNC.
利益披露 Disclosure
A. Jangholi, None..
B. Basnayake, None..
O. Breik, None..
S. Vasani, None..
C. Puyadeera, None.