肾癌源性Exosome通过TGF-β/Smad通路靶向调控Eomes诱导肿瘤浸润NK细胞向ILC1样细胞转化的机制研究

NK cells are important anti-tumor immune cells, their normal functions are inhibited in the tumor microenvironment. The mechanism is not yet clear. A recent study published by the Journal of Nature Immunology revealed a new mechanism that tumors achieved immune escape by inducing tumor-infiltrating NK cells into ILC1-like cells. Combined with the previous studies we found that renal tumor infiltrating NK cells had the characteristics of ILC1-like cells and renal tumor cell-derived exosome induced classical NK cells change to ILC1-like cells. We propose the scientific hypothesis that renal cell carcinoma-derived exosome targets Eomes via TGF-β/Smad signaling pathway to induce NK cell change to ILC1-like cell, allowing tumor cells to evade immune surveillance. In this study, we determined to explore the role of renal cancer-derived exosome on NK cells and its mechanism from there aspects, that is, clinical specimens, cells and animals, using the renal cell infiltrating NK cells as the entry point, using flow cytometry, CRISPR/Cas9, Cre-loxp, CHIP, GST-Pull Down, luciferase reporter gene detection and other technologies, trying to provide an important basis and clue for immunotherapy of tumors.

NK细胞是机体重要的抗肿瘤免疫细胞，其正常功能在肿瘤微环境中受到抑制，具体机制尚不清楚。近期自然免疫杂志发布研究揭示了肿瘤通过诱导肿瘤浸润NK细胞向ILC1样细胞转化从而实现免疫逃逸这一新机制。结合我们前期研究发现肾癌浸润NK细胞具有ILC1样细胞的特征及肾癌来源exosome诱导经典NK细胞发生ILC1样细胞样转化，我们提出科学假设：肾癌来源exosome通过TGF-β/Smad信号通路靶向Eomes诱导NK细胞向ILC1样细胞转化，使肿瘤细胞逃避免疫监视。本项研究从临床标本、细胞和动物三个层面，以肾癌浸润NK细胞为切入点，利用流式技术、CRISPR/Cas9、Cre-loxp、CHIP、GST-Pull Down、荧光素酶报告基因检测等技术探索肾癌exosome对NK细胞作用及其诱导NK细胞向ILC1样细胞转化的机制，为肿瘤的免疫治疗提供重要依据和线索。

NK细胞是机体重要的抗肿瘤免疫细胞，其正常功能在肿瘤微环境中受到抑制，具体机制尚不清楚。在前期的研究中我们发现肾癌来源的外泌体可以抑制NK细胞的激活性受体表达，增加抑制性受体表达，降低NK细胞的杀伤功能，诱导肿瘤的免疫耐受。在本项目中，我们将进一步鉴定肾癌浸润NK细胞的表型特征，研究其在肿瘤微环境功能变化情况及机制。我们首次发现肾癌微环境中肿瘤浸润NK细胞发生ILC1样细胞转化，并且肿瘤外泌体介导这一过程。我们使用单细胞测序及生物信息学分析深入探讨肿瘤浸润NK细胞发生亚群转化及免疫耐受的具体机制。本研究不仅阐明肿瘤细胞与TINK细胞交叉互动的关系及分子机制，而且为肿瘤的免疫治疗提供重要依据和线索，对提高患者预后及开发新型免疫治疗方法具有重要意义。