PO.CL05.10 · 临床研究
miR-150在乳腺癌中协调免疫激活和淋巴细胞迁移
miR-150 orchestrates immune activation and lymphocyte trafficking in breast cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：已知肿瘤浸润淋巴细胞（TIL）与治疗应答相关，进而带来更好的生存；然而，大多数乳腺癌（BC）几乎没有任何TIL。因此，发现可靶向的TIL浸润新机制有望具有重大临床意义。据报道，miR-150促进细胞增殖和迁移，但其在TIL浸润中的作用尚不清楚。本研究的目的是探讨肿瘤miR-150表达在BC患者中的作用、临床相关性及其对TIL的招募。
方法：对来自大型独立队列——癌症基因组图谱（TCGA）和乳腺癌国际分子分类联盟（METABRIC）的共计1,961例乳腺癌患者进行了计算机分析（in silico）。使用MDA-MB231和BT-549 BC细胞系以及Jurkat淋巴细胞系的体外实验重复了三次，以确保严谨性和可重复性。
结果：正如预期，miR-150表达与Nottingham分级相关，并且在两个队列中三阴性亚型中均更高（所有p<0.001）。另一方面，通过基因集变异分析，miR-150表达不仅富集MTORC1和KRAS信号通路，还富集多个免疫相关的Hallmark基因集，包括IFN-gamma、TNF-alpha、IL-2、IL-6、IFN-alpha、同种异体移植排斥和炎症反应（所有Spearman系数r>0.50且p<0.01）。在TCGA队列中，高miR-150表达与淋巴细胞浸润和TCR-Shannon显著相关。高miR-150表达与CD8+、CD4+记忆T细胞和树突状细胞的高浸润相关，并且在两个队列中均与溶细胞活性强相关（r=0.824和0.786，均p<0.01），所有这些均提示与免疫细胞浸润和免疫反应存在密切关系。与此一致，高miR-150表达与总生存的改善相关（p<0.001，p=0.030），尤其是在ER阳性/HER2阴性患者中。令人惊讶的是，miR-150模拟物过表达在MDA-MB231或BT-549 BC细胞系中均未促进细胞增殖、迁移或侵袭。然而，miR-150模拟物在MB231或BT-549细胞中的过表达显著增强了Jurkat细胞的迁移强度，这通过Transwell侵袭实验得以证实。此外，miR-150模拟物在MDA-MB231细胞中的过表达富集了细胞增殖相关的基因集：E2F靶点、G2M检查点，以及多个免疫相关基因集，包括IFN-gamma、TNF-alpha、IL-2、IL-6、同种异体移植排斥和炎症反应。
结论：患者乳腺癌中的miR-150表达激发免疫反应，将淋巴细胞吸引到肿瘤微环境中，并与总生存相关。
查看英文原文 English abstract
Background: Tumor-infiltrating lymphocytes (TILs) are known to relate with response to treatments followed by better survival; however, majority of breast cancer (BC) hardly have any TILs. Thus, discovery of targetable novel mechanism of TIL infiltration is expected to have a major clinical implication. MiR-150 has been reported to promote cell proliferation and migration, but its role in TIL infiltration is not known. The aim of this study is to investigate the role and clinical relevance of tumor miR-150 expression and attraction of TILs in BC patients.
Methods: In silico analyses was conducted on total of 1,961 breast cancer patients from large independent cohorts, The Cancer Genome Atlas (TCGA) and the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC). In Vitro experiments using MDA-MB231 and BT-549 BC cell lines and Jurkart lymphocyte cell line, were repeated three times to ensure rigor and reproducibility.
Results: As expected, miR-150 expression correlated with Nottingham grade and was higher in triple negative subtype in both cohorts (all p<0.001). On the other hand, miR-150 expression not only enriched MTORC1 and KRAS signaling, but also multiple immune-related Hallmark gene sets including IFN-ϒ, TNF-alpha, IL-2, IL-6, IFN-alpha, allograft rejection, and inflammatory response by gene set variant analysis (all Spearman's coefficient r>0.50 and p<0.01). High miR-150 expression significantly correlated with lymphocyte infiltration and TCR-Shannon in TCGA cohort. High miR-150 expression was associated with high infiltration of CD8+, CD4+ memory T cells and dendritic cells, and was strongly correlated with cytolytic activity consistently in both cohorts (r=0.824 and 0.786, both p<0.01), all suggesting strong relationship with immune cell infiltration and immune response. In agreement, high MiR-150 expression was associated with improved overall survival (p<0.001, p=0.030), particularly in ER-positive/HER2-negative patients. Surprisingly, mimic overexpression of miR-150 did not promote cell proliferation, migration nor invasion neither in MDA-MB231 or BT-549 BC cell lines. However, mimic overexpression of miR-150 in either MB231 or BT-549 cells significantly increased the migration intensity of Jurkart cells demonstrated by Transwell invasion assay. Further, mimic overexpression of miR-150 in MDA-MB231 cells enriched cell proliferation-related gene sets; E2F Targets, G2M checkpoint, as well as multiple immune-related gene sets including IFN-ϒ, TNF-alpha, IL-2, IL-6, allograft rejection and inflammatory response.
Conclusions: MiR-150 expression in patients' breast cancer evoke immune response, attracts lymphocytes to tumor microenvironment and is associated with overall survival.
利益披露 Disclosure
M. Oshi, None.