长非编码RNA NKILA和IRIE 调控乳腺癌免疫耗竭的机制研究

We have found before that the interaction between tumor associated macrophages and other stromal cells can promote breast cancer cells metastasis. Our pre-experiments showed that tumor associated macrophages after breast cancer preoperative chemotherapy have high expression of immune suppression molecule, such as FasL, which induces tumor specific T cells apoptosis. We have found that lncRNA NKILA and IRIE are upregulated in immune exhausting T cells and macrophages respectively via lncRNA array screening. Moreover, NKILA targets NFKB pathway and IRIE targets interferon pathway to induce breast cancer immune suppression. On this basis, our research plan to analyse the patients clinical samples and establish humanized mice and knockout mice to investigate the role of lncRNA in regulating immune exhaustion of breast cancer stromal cells. In addition, we will investigate the mechanism of differential lncRNA expression through bioinformatics analysis and luciferase reporter assay. Furthermore, we will reveal the molecular mechanism of the interaction between lncRNA and inflammatory signaling proteins via RNA pulldown and RIP. Our study will shed light on the molecular mechanism of the immune suppression of malignant cancer and help to provide novel molecular targets for therapies against tumor inflammatory microenvironment.

我们前期研究发现微环境中的巨噬细胞和其他间质细胞的互动促进乳腺癌转移。我们预实验提示术前化疗后的肿瘤浸润巨噬细胞高表达Fas-L等免疫耗竭分子，诱导肿瘤特异T细胞凋亡。通过高通量筛选发现NKILA和IRIE这两条长非编码RNA分别在免疫耗竭的T细胞和巨噬细胞差异表达，并分别通过靶向NFKB和干扰素通路诱导乳腺癌免疫抑制。在此基础上，本课题拟通过病人组织标本分析、人源化小鼠和基因敲除小鼠等模型进一步阐明长非编码RNA调控间质细胞免疫耗竭的作用；通过生物信息学和启动子报告载体构建阐述长非编码RNA差异表达的机制；通过RNA pulldown和RIP等技术揭示长非编码RNA与炎症信号蛋白相互作用的分子机理。本研究将有助于揭示恶性肿瘤免疫耗竭的分子机制，并为探索针对免疫细胞的长非编码RNA发展新型免疫靶向治疗药物提供新思路。

项目的研究方向为乳腺癌肿瘤微环境。主要学术成果：1.通过单细胞测序发现一系列具有治疗价值的膜蛋白靶点；2. 阐明抗肿瘤免疫在单克隆抗体治疗中的作用；3. 揭示肿瘤微环境中代谢时空变化。发表SCI文章12篇，其中IF大于10分11篇，包括Cell本刊4篇，Nature本刊1篇, Nature Immunology, Nature Cell Biology, Cell Research, Nature Communications, Clinical Cancer Research等。后续获得国自然重大研究计划、科技部重点研发计划等项目支持。