Exploring the oncoproteomic response of human prostate cancer to therapeutic radiation using data-independent acquisition (DIA) mass spectrometry
Exploring the oncoproteomic response of human prostate cancer to therapeutic radiation using data-independent acquisition (DIA) mass spectrometry
复制标题
DOI：
10.1002/pros.23500
复制
发表时间：
2018-06-01
期刊：
影响因子：
2.8
通讯作者：
Williams, Scott G.
中科院分区：
文献类型：
--
作者：
Keam, Simon P.;Gulati, Twishi;Williams, Scott G.
前言前列腺癌（PCa）放射抵抗的发展是一个重要的临床问题，目前仍缺乏个性化的分子特征。本研究的目的是建立一个平台，描述了早期癌蛋白质组反应的人前列腺组织放射治疗（RT）使用前瞻性的人体tissuecohol.MethodsFresh和固定的经会阴活检8名男子与临床局部肿瘤之前和14天后，一个单一的分数的高剂量率近距离放射治疗。使用优化的蛋白质提取管道和随后的数据独立采集质谱（DIA-MS）实现定量蛋白质分析。本体论分析被用来确定丰富的功能途径，进一步询问的候选人在福尔马林固定的石蜡包埋的组织活检从另外5 patients.ResultsWe新鲜组织活检的平均覆盖率为5660蛋白质，观察到的主要辐射后的变化是一个增加的水平之间共有49个蛋白质表现出丰富的变化。许多这些变化在患者之间存在差异，通常是前列腺癌组织，表现出高度的异质性。本体论分析揭示了三种免疫途径的蛋白质激活级联的富集：体液免疫应答、白细胞介导的免疫和补体激活。这些主要与细胞外间隙相关。我们验证了显着的表达差异，在20%和61%之间的这些候选人使用单独的固定组织队列，并建立其可行性作为实验组织资源，通过获取定量数据的平均值为5152蛋白每patient.DiscussionIn这项前瞻性研究，我们已经建立了一个敏感和可靠的癌蛋白质组学管道的新鲜和福尔马林固定的人前列腺癌组织的分析。我们确定了多种已知的辐射反应途径，并建立了一个强大的候选人和途径数据库，目前与RT无关。这些信息可能有利于个性化治疗和潜在的预测性生物标志物的发展。
IntroductionThe development of radioresistance in prostate cancer (PCa) is an important clinical issue and is still largely uninformed by personalized molecular characteristics. The aim of this study was to establish a platform that describes the early oncoproteomic response of human prostate tissue to radiation therapy (RT) using a prospective human tissue cohort.MethodsFresh and fixed transperineal biopsies from eight men with clinically localized tumors were taken prior to and 14 days following a single fraction of high-dose-rate brachytherapy. Quantitative protein analysis was achieved using an optimized protein extraction pipeline and subsequent data-independent acquisition mass spectroscopy (DIA-MS). Ontology analyses were used to identify enriched functional pathways, with the candidates further interrogated in formalin-fixed paraffin-embedded tissue biopsies from five additional patients.ResultsWe obtained a mean coverage of 5660 proteins from fresh tissue biopsies; with the principal post-radiation change observed being an increase in levels amongst a total of 49 proteins exhibiting abundance changes. Many of these changes in abundance varied between patients and, typically to prostate cancer tissue, exhibited a high level of heterogeneity. Ontological analysis revealed the enrichment of the protein activation cascades of three immunological pathways: humoral immune response, leukocyte mediated immunity and complement activation. These were predominantly associated with the extracellular space. We validated significant expression differences in between 20% and 61% of these candidates using the separate fixed-tissue cohort and established their feasibility as an experimental tissue resource by acquiring quantitative data for a mean of 5152 proteins per patient.DiscussionIn this prospective study, we have established a sensitive and reliable oncoproteomic pipeline for the analysis of both fresh and formalin-fixed human PCa tissue. We identified multiple pathways known to be radiation-responsive and have established a powerful database of candidates and pathways with no current association with RT. This information may be beneficial in the advancement of personalized therapies and potentially, predictive biomarkers.