TAMs促进至虚肿瘤微环境形成致结直肠癌干细胞毒根深藏的机制研究

Colorectal cancer (CRC) recruits macrophages and induces the M2-like phenotype of tumor-associated macrophages (TAMs). TAMs make the healthy-Qi increasingly deficient and promote tumor microenvironment of extreme deficiency. The most deficient place stores the most toxic pathogen. The extreme deficient macroenvironment sculptures the stemness of CRC cells to induce colorectal cancer stem cells (CCSCs). Our previous works found that transforming growth factor β1 (TGF-β1) activated Sox4 and LASP1 and induced epithelial-mesenchymal transition (EMT). Shenling Baizhu San downregulated the number of TAMs in mice with colitis-associated CRC. Tonifying-Pi strengthening healthy-Qi and removing cancerous toxin lowered TGF-β1 level of CRC patients. We thus propose that TAMs maintains its M2-like phenotype through TGF-β1/LASP1 pathway to form the tumor microenvironment of extreme deficiency and activate Wnt, Sox and EMT procedure to root colorectal cancer stem cells. Pathogenesis is key for syndrome differentiation and treatment. Clinical test will be adopted to observe the correlation between the M2 phenotype of TAMs and CCSCs. Transgenic mice hybridization and human-derived CCSCs transplantation will be used to analyze the effects of tonifying-Pi strengthening healthy-Qi and removing cancerous toxin and the role of TGF-β1/LASP1 in regulating TAMs phenotype and CCSCs stemness. The reciprocal relationship between TAMs and CCSCs will be elucidated with cell sorting. The classic quotation “where is the deficiency of healthy-Qi, where is the site for toxic pathogen" will be reinvigorated by the new meanings.

结直肠癌（CRC）募集肿瘤相关巨噬细胞（TAMs）参与形成肿瘤微环境。M2型TAMs使脾虚免疫微环境的正气愈加亏虚，形成至虚微环境。“最虚之处，便是容邪之处”，至虚微环境是诱导干性基因表达产生CRC干细胞（CCSCs）之本。我们前期发现，TGF-β1活化Sox4，诱导LASP1活化发生EMT；参苓白术散下调结肠炎相关癌小鼠TAMs数量；健脾扶正解毒降低CRC病人TGF-β1水平。因此，我们提出假说：TAMs通过TGF-β1/LASP1维持其M2表型而促进至虚微环境形成，激活Wnt、Sox及EMT进程诱导CCSCs形成毒根。治病必求于本。拟临床观察TAMs的M2表型与CCSCs相关性；应用转基因小鼠杂交及人源CCSCs移植等多种模型，分析健脾扶正解毒调节TAMs表型及CCSCs干性的机制；细胞解析TAMs与CCSCs之间的相互作用及调控机制，为“邪之所凑，其气必虚”的经典论述注入新的内涵。

结直肠癌（CRC）募集肿瘤相关巨噬细胞（TAMs）参与形成肿瘤微环境。M2型TAMs使脾虚免疫微环境的正气愈加亏虚，形成至虚免疫微环境，而至虚微环境可能是诱导干性基因生成CRC肿瘤干细胞（CSCs）之本。我们的研究发现，TGFβ诱导的上皮间质转化（EMT）进程增加CRC肿瘤干细胞的干性，高迁移率族蛋白B2（HMGB2）核转位启动EMT，并上调干性基因八聚体结合转录因子4（OCT4）表达；TGFβ1促进肿瘤转移微环境纤维连接蛋白（FN）和胶原沉积，增加细胞外基质（ECM）重构，并诱导肿瘤相关巨噬细胞（TAMs）由M1型向M2型迁移，促进CRC的侵袭与转移；扶正健脾之参苓白术散加减方减少TGFβ1分泌，降低CSCs干性，下调腺瘤缺氧诱导因子-1α（HIF-1α）的表达，减少CD4+CD25+FoxP3+调节性T细胞（Tregs）向癌巢的募集，抑制CRC生长与转移。.我们的结果阐明了TGFβ诱导TAMs由M1向M2表型分化产生正虚肿瘤微环境机制；发现EMT和HMGB2核转位上调CSCs干性并促进转移。基于以上研究内容，提出肿瘤干细胞Niche 是容许CSCs异常增殖的特化肿瘤微环境，CSCs 居于正气亏虚肿瘤微环境的至虚之处，元气益虚而毒根愈固，凝炼肿瘤微环境中“最虚之处，便是容邪之处”的内涵，提出了肿瘤干细胞是“至虚之处，毒根深藏”的观点，并建议通过扶正来干预 CSCs及其赖以生存之niche改善微环境的治疗策略。项目通过探索了肿瘤微环境中“最虚之处，便是容邪之处”的“正虚”与“癌毒”交互为患的中医辩证关系，为临床应用扶正解毒、化痰祛瘀法调节肿瘤干细胞niche奠定理论基础并提供实验依据，具有较为广阔的临床应用前景。.依托本项目共发表SCI收录期刊发表文章9篇，中文核心期刊文章5篇（均标注项目号），培养博士后1人、博士生3人、硕士生3人，获授权专利1项，完成项目表述的研究内容。