Sougiannis Alexander T, Taylor Harrison B, Zambrzycki Stephen C, Conroy Lindsey, Strubler Rachel, Edge Christin, Drake Richard R, Wallace Kristin, Allison Derek, Lee Eun, Sun Ramon C, Angel Peggi M
College of Medicine, Medical University of South Carolina, Charleston, SC 29403, USA.
Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29403, USA.
Oncol Lett. 2025 Jun 26;30(3):413. doi: 10.3892/ol.2025.15159. eCollection 2025 Sep.
Emerging evidence reports that regulation of the extracellular matrix influences the progression of colorectal cancer (CRC). The present study investigated regulation of the extracellular matrix proteome in colorectal malignancy within a high-risk Appalachian population compared with non-Appalachian populations. A targeted mass spectrometry imaging proteomic method directed at collagen regulation was used. Tissue microarrays (TMAs) comprising of matched CRC with adjacent normal to tumor (NAT) from 45 patients were constructed into 86 samples to evaluate the extracellular matrix proteome (ECM). A total of five specific peaks were discovered to differ between NAT and tumor with high sensitivity and specificity by receiver operating characteristic (AUROC) ≥0.7, Wilson/Brown P<0.0002. Evaluation of patient TMA cores showed increased levels of combined ECM peptides in advanced stage Appalachian CRC (III + IV) compared with early staged CRC (I + II) (AUROC 0.8595; 95% confidence interval, 0.8190-0.8999; Wilson/Brown P<1.0×10), contrasting with the non-Appalachian tumors, which showed a decreased ability to discriminate between early and late stage (AUROC 0.6618; 95% confidence interval, 0.6126-0.7110; Wilson/Brown P<1.0×10). Comparison of advanced stage CRCs between Appalachian and non-Appalachian populations showed high sensitivity and specificity in distinguishing the populations (AUROC 0.7612; 95% confidence interval, 0.7109-0.8114; Wilson/Brown P<3.0×10). History of smoking, sex and tumor origin location did not show significant ability to distinguish by AUROC. A combination of high mass resolution, high mass accuracy spatial proteomics and sequencing proteomics by liquid chromatography coupled to tandem mass spectrometry revealed that fibrillar collagens were spatially regulated within the CRC tumor microenvironment. Fibrillar collagen post-translational modifications of hydroxylated proline revealed distinct spatial separation based on the presence of a number of hydroxylated proline sites. The present study highlighted that the targeted mass spectrometry imaging of the ECM proteome may provide new insight and novel predictive tools for understanding CRC, particularly among Appalachian patients.
新出现的证据表明，细胞外基质的调控会影响结直肠癌（CRC）的进展。本研究调查了高危阿巴拉契亚人群与非阿巴拉契亚人群相比，结直肠恶性肿瘤中细胞外基质蛋白质组的调控情况。采用了一种针对胶原蛋白调控的靶向质谱成像蛋白质组学方法。将来自45名患者的配对结直肠癌组织与相邻正常组织（NAT）构建成组织微阵列（TMA），共86个样本，以评估细胞外基质蛋白质组（ECM）。通过受试者工作特征曲线（AUROC）≥0.7以及Wilson/Brown P<0.0002，共发现五个特定峰在NAT和肿瘤之间存在差异，具有高灵敏度和特异性。对患者TMA核心的评估显示，与早期结直肠癌（I + II期）相比，晚期阿巴拉契亚结直肠癌（III + IV期）中ECM肽的组合水平升高（AUROC 0.8595；95%置信区间，0.8190 - 0.8999；Wilson/Brown P<1.0×10），而非阿巴拉契亚肿瘤在区分早期和晚期方面的能力下降（AUROC 0.6618；95%置信区间，0.6126 - 0.7110；Wilson/Brown P<1.0×10）。阿巴拉契亚人群与非阿巴拉契亚人群晚期结直肠癌的比较显示，在区分人群方面具有高灵敏度和特异性（AUROC 0.7612；95%置信区间，0.7109 - 0.8114；Wilson/Brown P<3.0×10）。吸烟史、性别和肿瘤起源位置通过AUROC未显示出显著的区分能力。通过液相色谱 - 串联质谱联用的高质量分辨率、高质量精度空间蛋白质组学和测序蛋白质组学相结合的方法表明，纤维状胶原蛋白在结直肠癌肿瘤微环境中受到空间调控。基于多个羟基化脯氨酸位点的存在，羟基化脯氨酸的纤维状胶原蛋白翻译后修饰显示出明显的空间分离。本研究强调，ECM蛋白质组的靶向质谱成像可能为理解结直肠癌提供新的见解和新的预测工具，特别是在阿巴拉契亚患者中。
