PO.CH02.01 · 化学
用于葡萄膜黑色素瘤预后分层的房水蛋白绝对定量
Absolute quantification of aqueous humor proteins for prognostic stratification in uveal melanoma
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：葡萄膜黑色素瘤（UM）是成人最常见的原发性眼内恶性肿瘤，具有较高的转移进展风险，尤其是在具有高危基因表达谱或晚期临床分期的肿瘤中。直接肿瘤活检可提供有价值的预后信息，但存在操作风险，凸显了对微创方法以捕获肿瘤来源生物标志物的需求。房水（AH）含有眼内肿瘤释放的可检测分子物质，但此前尚未对其中的蛋白质生物标志物进行绝对定量。本研究旨在建立UM首个校准的AH蛋白质组图谱，并识别与分子分类和疾病分期相关的蛋白质生物标志物。
方法：在斑块近距离放疗前采集了70只UM眼的AH样本。细针肿瘤穿刺提供了基因表达谱（GEP1、GEP2）和PRAME状态。所有样本均接受了基于NGS的高重邻近延伸测定（PEA），其中27份有剩余体积的样本接受了针对92种蛋白的基于qPCR的PEA。基于66种重叠蛋白建立的回归模型能够在整个队列中外推绝对pg/mL浓度。对中位浓度>5 pg/mL的蛋白评估其在GEP分类、PRAME状态和AJCC分期方面的差异。使用Ingenuity Pathway Analysis识别通路富集和上游调控因子。
结果：23种蛋白以临床可定量浓度存在。GEP1/PRAME-肿瘤中观察到最低的蛋白水平，随AJCC分期进展呈逐步升高。在IV期疾病中，CXCL8（p=0.00024）、IL10（p=0.00049）和PDCD1（p=0.00094）显著升高。通路分析显示免疫和肿瘤相关信号通路富集。VEGFA（z=3.1）和CCL2（z=1.7）被识别为预测的上游调控因子，并在晚期肿瘤中升高。
结论：本研究提供了UM中AH蛋白的首个绝对定量图谱，并识别了一组与肿瘤生物学、分子分类和分期相关的23种临床可检测生物标志物。这些发现使AH蛋白质组学成为一个能够捕获UM中分期相关免疫活性并支持标准化预后分层生物标志物开发的微创液体活检平台。
查看英文原文 English abstract
Background: Uveal melanoma (UM) is the most common primary intraocular malignancy in adults and carries a high risk of metastatic progression, particularly in tumors with high-risk gene expression profiles or advanced clinical stage. Direct tumor biopsy provides valuable prognostic information but carries procedural risk, highlighting the need for minimally invasive approaches to capture tumor-derived biomarkers. Aqueous humor (AH) contains detectable molecular material released by intraocular tumors, yet absolute quantification of protein biomarkers has not previously been performed. This study aims to establish the first calibrated AH proteome map for UM and to identify protein biomarkers associated with molecular classification and disease stage.
Methods: AH samples from 70 UM eyes were collected before plaque brachytherapy. Fine needle tumor aspiration provided gene expression profiling (GEP1, GEP2) and PRAME status. All samples underwent NGS based high-plex proximity extension assay (PEA), and 27 samples with remaining volume underwent qPCR-based PEA targeting 92 proteins. Regression models derived from 66 overlapping proteins enabled extrapolation of absolute pg/mL concentrations across the full cohort. Proteins with median concentrations >5 pg/mL were assessed for differences by GEP class, PRAME status, and AJCC stage. Pathway enrichment and upstream regulators were identified using Ingenuity Pathway Analysis.
Results: Twenty-three proteins were present at clinically quantifiable concentrations. The lowest protein levels were observed in GEP1/PRAME- tumors, with progressive increases seen across advanced AJCC stages. Significant elevations were noted in Stage IV disease for CXCL8 (p=0.00024), IL10 (p=0.00049), and PDCD1 (p=0.00094). Pathway analysis demonstrated enrichment in immune and tumor-associated signaling pathways. VEGFA (z=3.1) and CCL2 (z=1.7) were identified as predicted upstream regulators and were elevated in advanced-stage tumors.
Conclusions: This study provides the first absolute quantitative map of AH proteins in UM and identifies a panel of 23 clinically detectable biomarkers associated with tumor biology, molecular classification, and stage. These findings position AH proteomics as a minimally invasive liquid biopsy platform capable of capturing stage-linked immune activity in UM and supporting the development of standardized biomarkers for prognostic stratification.
利益披露 Disclosure
D. Liang, None..
E. Huang, None..
C. Peng, None..
J. Berry, None..
L. Xu, None.