本研究的目的是使用质谱分析来自葡萄膜黑色素瘤 (UM) 患者的液体玻璃体活检的蛋白质表达，以确定预后生物标志物、信号通路和治疗靶点。玻璃体活检是从一项试点研究中的两个队列中收集的：比较对照具有视网膜前膜的眼睛（ERM；n = 3）和具有 UM 的测试眼睛（n = 8）。使用液相色谱-串联质谱 (LC-MS/MS) 分析样品。将鉴定出的蛋白质与来自靶向多重 ELISA 蛋白质组学平台的数据进行比较。在 UM 玻璃体中总共检测到 69 种显着升高的蛋白质，包括 LYVE-1。 LC-MS/MS 鉴定出 UM 玻璃体中 62 种显着上调的蛋白质，这些蛋白质以前未通过 ELISA 鉴定出。通过肿瘤分子分类（基因表达谱 [GEP] 和黑色素瘤中优先表达的抗原 [PRAME]）对差异蛋白表达进行分析，进一步鉴定了与这些分类相关的蛋白。患有高危 GEP 肿瘤的患者表现出 HGFR 玻璃体表达升高（倍数变化 [FC] = 2.66E   03，P 值 = 0.003）和 PYGL（FC = 1.02E  04，P = 1.72E-08）。 PRAME 阳性肿瘤患者玻璃体中 ENPP-2 (FC = 3.21，P = 0.04)、NEO1 (FC = 2.65E   03，P = 0.002) 和 LRP1 (FC = 5.59E  02，P 值 = 0.01) 表达升高。 IGF 调节效应器的代表性很高（P 值 = 1.74E-16）。跨平台分析验证了通过 ELISA 和 LC-MS/MS 鉴定的七种蛋白质。液体活检的蛋白质组学分析可以提供支持肿瘤活检基因表达的预后信息。在同一患者样本中使用多个蛋白质检测平台可以提高候选生物标志物检测的灵敏度，并可以精确表征玻璃体蛋白质组。

The purpose of this study was to profile protein expression liquid vitreous biopsies from patients with uveal melanoma (UM) using mass spectrometry to identify prognostic biomarkers, signaling pathways, and therapeutic targets.Vitreous biopsies were collected from two cohorts in a pilot study: comparative control eyes with epiretinal membranes (ERM; n = 3) and test eyes with UM (n = 8). Samples were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Identified proteins were compared to data from a targeted multiplex ELISA proteomics platform.A total of 69 significantly elevated proteins were detected in the UM vitreous, including LYVE-1. LC-MS/MS identified 62 significantly upregulated proteins in UM vitreous that were not previously identified by ELISA. Analysis of differential protein expression by tumor molecular classification (gene expression profiling [GEP] and preferentially expressed antigen in melanoma [PRAME]) further identified proteins that correlated with these classifications. Patients with high-risk GEP tumors displayed elevated vitreous expression of HGFR (fold-change [FC] = 2.66E + 03, P value = 0.003) and PYGL (FC = 1.02E + 04, P = 1.72E-08). Patients with PRAME positive tumors displayed elevated vitreous expression of ENPP-2 (FC = 3.21, P = 0.04), NEO1 (FC = 2.65E + 03, P = 0.002), and LRP1 (FC = 5.59E + 02, P value = 0.01). IGF regulatory effectors were highly represented (P value = 1.74E-16). Cross-platform analysis validated seven proteins identified by ELISA and LC-MS/MS.Proteomic analysis of liquid biopsies may provide prognostic information supporting gene expression of tumor biopsies. The use of multiple protein detection platforms in the same patient samples increases the sensitivity of candidate biomarker detection and allows for precise characterization of the vitreous proteome.