Ning Wu, Jia Wenqing, Ning Jingyuan, Zhou Lei, Li Zongze, Zhang Lin, Song Xin
Department of General Surgery, China-Japan Friendship Hospital, Beijing, China.
Institute of Clinical Medicine, China-Japan Friendship Hospital, Beijing, China.
Front Immunol. 2025 Jun 12;16:1628005. doi: 10.3389/fimmu.2025.1628005. eCollection 2025.
The pronounced heterogeneity of colorectal cancer (CRC) significantly impacts patient prognosis and therapeutic response, making elucidation of its molecular mechanisms critical for developing precision treatment strategies. This study aimed to systematically characterize tumor cell heterogeneity and explore its clinical implications.
Five single-cell RNA sequencing cohorts were integrated (comprising 70 CRC samples and 164,173 cells) to systematically analyze tumor cell heterogeneity. Unsupervised clustering analysis based on VEGFR+ tumor cell signature genes was used to stratify CRC patients. Key molecular mechanisms were validated through in vitro cellular experiments, in vivo animal models, molecular docking, and dynamics simulations.
The analysis successfully identified five distinct tumor cell subtypes, with the VEGFR+ subtype exhibiting marked epithelial-mesenchymal transition (EMT) activation signatures and strong association with metastasis and poor clinical outcomes. Based on VEGFR+ signature genes, CRC patients were stratified into three subgroups: C1 (metabolically active), C2 (proliferative), and C3 (invasive), with the C3 subtype demonstrating high metastatic potential, stem-like properties, and an immunosuppressive microenvironment, along with a five-year survival rate below 50%. Mechanistic investigations identified HOXC6 as a key driver of the C3 subtype, with HOXC6 knockout significantly suppressing CRC cell proliferation, migration, and invasion. Furthermore, molecular docking revealed that the targeted agent abemaciclib effectively binds HOXC6, with both cellular and animal experiments confirming its ability to inhibit CRC cell functions and significantly reduce tumor burden in nude mice.
This study establishes the first single-cell-resolution molecular classification system for CRC, delineates the mechanistic link between EMT subtypes and metastatic progression, and identifies HOXC6 as a novel therapeutic vulnerability. These findings provide a translational foundation for precision oncology and offer new rationale for precision diagnosis and treatment of colorectal cancer.
结直肠癌（CRC）显著的异质性对患者预后和治疗反应有重大影响，因此阐明其分子机制对于制定精准治疗策略至关重要。本研究旨在系统地表征肿瘤细胞异质性并探索其临床意义。
整合了五个单细胞RNA测序队列（包含70个CRC样本和164,173个细胞）以系统分析肿瘤细胞异质性。基于VEGFR +肿瘤细胞特征基因的无监督聚类分析用于对CRC患者进行分层。通过体外细胞实验、体内动物模型、分子对接和动力学模拟验证关键分子机制。
分析成功识别出五种不同的肿瘤细胞亚型，其中VEGFR +亚型表现出明显的上皮-间质转化（EMT）激活特征，并与转移和不良临床结果密切相关。基于VEGFR +特征基因，CRC患者被分为三个亚组：C1（代谢活跃）、C2（增殖）和C3（侵袭），C3亚型具有高转移潜力、干细胞样特性和免疫抑制微环境，五年生存率低于50%。机制研究确定HOXC6是C3亚型的关键驱动因素，敲除HOXC6可显著抑制CRC细胞的增殖、迁移和侵袭。此外，分子对接显示靶向药物阿贝西利可有效结合HOXC6，细胞和动物实验均证实其能够抑制CRC细胞功能并显著减轻裸鼠的肿瘤负担。
本研究建立了首个用于CRC的单细胞分辨率分子分类系统，描绘了EMT亚型与转移进展之间的机制联系，并确定HOXC6是一种新的治疗靶点。这些发现为精准肿瘤学提供了转化基础，并为结直肠癌的精准诊断和治疗提供了新的理论依据。
