PO.CL05.09 · 临床研究
CCL3+肿瘤细胞通过IDE+ M2巨噬细胞和CCR3+ CAF在ESCC中协调免疫平衡与干性维持
CCL3+ tumor cells orchestrate immune equilibrium and stemness maintenance in ESCC via IDE+ M2 macrophages and CCR3+ CAFs
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
免疫调节在肿瘤发生与发展中发挥关键作用，但其潜在机制在很大程度上仍不清楚。通过构建小鼠模型追踪肿瘤免疫状态，并利用单细胞测序技术剖析不同阶段肿瘤细胞与肿瘤微环境之间的相互作用，我们发现ESCC肿瘤进展经历四个关键阶段：①免疫识别期（CXCL10/H2-K1抗原呈递信号增强）；②免疫平衡期（PD-1 T细胞/CD44休眠肿瘤细胞富集）；③免疫逃逸期（PD-L1+ EMT肿瘤细胞增殖）；④免疫抑制期（由M2 TAM/Treg主导）。细胞通讯分析显示，肿瘤内的细胞间相互作用主要发生于肿瘤细胞、巨噬细胞、成纤维细胞和T细胞之间，其中肿瘤细胞作为核心调控因素。对肿瘤细胞亚型进一步的GO分析显示富集于角蛋白强化型（Cnfn+）、自噬增强型（Plk5+）、免疫抑制型（Ccl3+）和转移增强型（Krt8+）。其中，Ccl3+肿瘤细胞在免疫平衡期高度富集，并与Ide M2巨噬细胞和Ccr3+ CAF相互作用。体外共培养研究证实，过表达Ccl3的肿瘤细胞可调节巨噬细胞向M2极化并促进T细胞耗竭；然而，在敲低巨噬细胞中的Ide基因后，这些功能不再改变。另一方面，当过表达Ccl3的肿瘤细胞与Ccr3敲除CAF及对照CAF共培养时，对照组显著增强Ccl3+肿瘤细胞的干性。这些研究证实，Ccl3+肿瘤细胞通过Ide M2巨噬细胞和Ccr3+成纤维细胞重塑肿瘤微环境，从而调控干性并在免疫平衡期存活。在体内，靶向Ccl3的中和抗体可有效抑制皮下肿瘤形成并增强免疫效应。本研究揭示了Ccl3在ESCC肿瘤免疫演变中的关键调控作用，为克服免疫治疗耐药和开发新型靶向疗法提供了重要的分子基础和潜在的临床策略。
查看英文原文 English abstract
Immune regulation plays a crucial role in tumor occurrence and development, yet the underlying mechanisms remain largely unclear. By constructing a mouse model to track tumor immune status and utilizing single-cell sequencing technology to dissect the interactions between tumor cells and the tumor microenvironment at different stages, we found that ESCC tumor progression undergoes four key phases: ① immune recognition phase (enhanced CXCL10/H2-K1 antigen presentation signaling); ② immune equilibrium phase (enrichment of PD-1 T cells/CD44 dormant tumor cells); ③ immune escape phase (proliferation of PD-L1+ EMT tumor cells); ④ immune suppression phase (dominated by M2 TAMs/Tregs). Cell communication analysis revealed that intercellular interactions in the tumor primarily occur among tumor cells, macrophages, fibroblasts, and T cells, with tumor cells serving as the core regulatory factor. Further GO analysis of tumor cell subtypes showed enrichment in keratin-reinforced type (Cnfn+), enhanced autophagy type (Plk5+), immunosuppressive type (Ccl3+), and metastasis-enhanced type (Krt8+). Among them, Ccl3+ tumor cells are highly enriched in the immune equilibrium phase and interact with Ide M2 macrophages and Ccr3+ CAFs. In vitro co-culture studies confirmed that tumor cells overexpressing Ccl3 can regulate macrophage polarization toward M2 and promote T cell exhaustion; however, these functions no longer change after knocking down the Ide gene in macrophages. On the other hand, when tumor cells overexpressing Ccl3 are co-cultured with Ccr3-knockout CAFs and control CAFs, the control group significantly enhances the stemness of Ccl3+ tumor cells. These studies confirm that Ccl3+ tumor cells remodel the tumor microenvironment through Ide M2 macrophages and Ccr3+ fibroblasts, thereby regulating stemness and surviving in the immune equilibrium phase. In vivo, neutralizing antibodies targeting Ccl3 effectively inhibit subcutaneous tumor formation and enhance immune effects. This study reveals the key regulatory role of Ccl3 in the immune evolution of ESCC tumors, providing an important molecular basis and potential clinical strategies for overcoming immunotherapy resistance and developing novel targeted therapies.
利益披露 Disclosure
B. Liu, None.