PO.IM02.01 · 免疫学
NOX2驱动的巨噬细胞重编程增强乳腺癌转移
NOX2-driven macrophage reprogramming enhances breast cancer metastasis
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
肿瘤相关巨噬细胞(TAM)被认为是乳腺癌进展的关键决定因素,然而塑造其免疫抑制表型的分子机制仍不完全清楚。髓系细胞的NOX2酶在髓系细胞中产生抗菌活性氧(ROS),但其对肿瘤微环境内巨噬细胞编程的潜在贡献在很大程度上未知。本研究旨在确定NOX2在调节M2样巨噬细胞极化以促进乳腺癌生长和播散中的潜在作用。对100例人乳腺肿瘤的单细胞RNA测序数据分析显示,NOX2在M2样TAM簇中选择性上调,并与免疫抑制性转录谱相关。与此一致,转录数据显示,来自Nox2缺陷小鼠的巨噬细胞表现出向促炎M1样状态的转变,以转录因子网络改变和炎症基因特征富集为标志。在体外,NOX2的药理学或遗传学抑制可阻断由集落刺激因子-1(CSF-1)(P=0.0006,n=4-11/组,单因素ANOVA)或乳腺癌来源条件培养基(P=0.02,n=4/组,Mann-Whitney检验)诱导的M2极化,提示NOX2活性是肿瘤驱动的巨噬细胞重编程所必需的。在机制上,我们的结果提示NOX2来源的ROS激活了NRF2依赖性转录通路,从而稳定了M2样表型。在两种不同的小鼠乳腺癌体内模型中,即原位EO771植入和基因工程MMTV-PyMT模型,Nox2的遗传学缺失导致M2样TAM丰度降低(P=0.006,n=4-7/组)、瘤内T细胞浸润增强(P=0.04,n=4-7/组),并显著抑制原发肿瘤生长(P=0.01,n=4-7/组)和转移播散(P=0.02,n=6-8/组,均采用Mann-Whitney检验)。对公共数据集的挖掘显示,M2相关标志物和NRF2靶基因的高表达与人乳腺癌较差的生存相关(CD206的P=0.02,HO-1的P=0.04,n=65,log-rank检验)。这些发现将NOX2确立为乳腺癌中免疫抑制性巨噬细胞可塑性的关键驱动因素,并突显NOX2-NRF2轴作为对抗促转移髓系编程的潜在治疗靶点。
查看英文原文 English abstract
Tumor-associated macrophages (TAMs) are considered key determinants of breast cancer progression, yet the molecular mechanisms shaping their immunosuppressive phenotypes remain incompletely understood. The NOX2 enzyme of myeloid cells generates antimicrobial reactive oxygen species (ROS) in myeloid cells, but its potential contribution to macrophage programming within the tumor microenvironment is largely unknown. This study aimed at identifying the potential role of NOX2 in regulating M2-like macrophage polarization for breast cancer growth and dissemination. Analysis of single-cell RNA sequencing data from 100 human breast tumors revealed selective upregulation of NOX2 in M2-like TAM clusters, associated with an immunosuppressive transcriptional profile. Consistently, transcriptional data showed that macrophages from Nox2 -deficient mice displayed a shift toward a pro-inflammatory, M1-like state, marked by altered transcription factor networks and enrichment of inflammatory gene signatures. In vitro , pharmacologic or genetic inhibition of NOX2 blocked M2 polarization induced by colony-stimulating factor-1 (CSF-1) (P=0.0006, n=4-11/group, one-way ANOVA) or breast cancer-derived conditioned media (P=0.02, n=4/group, Mann-Whitney test), implying that NOX2 activity is required for tumor-driven macrophage reprogramming. Mechanistically, our results suggested that NOX2-derived ROS activated NRF2-dependent transcriptional pathways that stabilized the M2-like phenotype. In two distinct murine breast cancer in vivo models, i.e. orthotopic EO771 implantation and the genetically engineered MMTV-PyMT model, genetic deletion of Nox2 entailed reduced abundance of M2-like TAMs (P=0.006, n=4-7/group), enhanced intratumoral T cell infiltration (P=0.04, n=4-7/group), and markedly suppressed primary tumor growth (P=0.01, n=4-7/group) and metastatic spread (P=0.02, n=6-8/group, Mann-Whitney test for all). Mining of public datasets revealed that high expression of M2-associated markers and NRF2 target genes correlated with inferior survival in human breast cancer (P=0.02 for CD206 and P=0.04 for HO-1, n=65, log-rank test). These findings position NOX2 as a key driver of immunosuppressive macrophage plasticity in breast cancer and highlight the NOX2-NRF2 axis as a potential therapeutic target to counteract metastasis-promoting myeloid programming.
利益披露 Disclosure
M. Kaya, None..
O. Johnsson, None..
N. Issdisai, None..
H. Söderberg, None..
I. Altinönder, None..
R. Kiffin, None..
X. Tekpli, None..
K. Hellstrand, None..
A. Martner, None.