MORF4L2通过GRHL2/MORF4L2/H4K12Ac/CSF1轴在三阴性乳腺癌中诱导免疫抑制微环境和免疫治疗耐药性。

MORF4L2 induces immunosuppressive microenvironment and immunotherapy resistance through GRHL2/MORF4L2/H4K12Ac/CSF1 axis in triple-negative breast cancer.

作者信息

Sui Xin-Yi, Cao Shuo-Wen, Song Xiao-Qing, Liu Xi-Yu, Chen Chao, Yan Qingya, Wang Zhi-Qing, Zhang Wen-Juan, Ma Lin-Xiaoxi, Jin Xi, Ma Ding, Xiao Yi, Wu Song-Yang, Xu Ying, Shao Zhi-Ming, Fan Lei

机构信息

Department of Breast Surgery, Fudan University Shanghai Cancer Center, Shanghai, China.

Key Laboratory of Breast Cancer in Shanghai, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China.

出版信息

Biomark Res. 2025 Jan 9;13(1):6. doi: 10.1186/s40364-024-00719-1.

DOI:10.1186/s40364-024-00719-1

PMID:39780291

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11715975/

Abstract

BACKGROUND

Although immunotherapy has achieved great progress in advanced triple-negative breast cancer (TNBC), there are still numerous patients who do not benefit from immunotherapy. Therefore, identification of the key molecule that induces immune escape and clarification of its specific mechanism in TNBC are urgently needed.

METHODS

In this research, single cell sequencing and bulk sequencing were conducted for biomarker screening. Immunohistochemistry, multiplex immunofluorescence, and orthotopic TNBC tumor model were applied in identifying the key molecule driving immune escape. At the mechanical level, RNA sequencing, in vitro co-culturing system, flow cytometry, Western blotting, ELISA, and real-time qPCR were carried out.

RESULTS

Mortality factor 4 like 2 (MORF4L2) expression is significantly up-regulated among patients who developed anti-PD1 resistance. MORF4L2 enhances anti-PD1 resistance by inducing the chemotaxis of macrophage infiltration and promoting their polarization towards the alternative activation phenotype (M2), thus creating an immunosuppressive microenvironment. Mechanistically, MORF4L2 actes as part of NuA4 histone acetyltransferase (HAT) complex, contributes to to histone 4 lysine 12 acetylation (H4K12Ac) and activates the downstream transcription of macrophage colony-stimulating factor (CSF1). CSF1 is secreted by tumor cells and binds to the macrophage-surface CSF1 receptor (CSF1R), which chemotactically converted and polarized macrophages to the M2 phenotype. Furthermore, we revealed that grainyhead like transcription factor 2 (GRHL2) could promote MORF4L2 transcription by binding to the MORF4L2 enhancer region. Notably, BLZ549, an inhibitor of CSF1R, restored the anti-PD1 sensitivity by blocking the GRHL2/MORF4L2/H4K12Ac/CSF1 axis.

CONCLUSIONS

GRHL2/MORF4L2/H4K12Ac/CSF1 axis plays an important role in anti-PD1 resistance. CSF1R inhibitors can reverse GRHL2/MORF4L2-mediated anti-PD1 resistance.

摘要

背景

尽管免疫疗法在晚期三阴性乳腺癌（TNBC）中取得了巨大进展，但仍有许多患者无法从免疫疗法中获益。因此，迫切需要鉴定诱导免疫逃逸的关键分子并阐明其在TNBC中的具体机制。

方法

在本研究中，进行了单细胞测序和批量测序以筛选生物标志物。应用免疫组织化学、多重免疫荧光和原位TNBC肿瘤模型来鉴定驱动免疫逃逸的关键分子。在机制层面，进行了RNA测序、体外共培养系统、流式细胞术、蛋白质免疫印迹、酶联免疫吸附测定和实时定量聚合酶链反应。

结果

在出现抗程序性死亡蛋白1（PD1）耐药的患者中，死亡因子4样蛋白2（MORF4L2）表达显著上调。MORF4L2通过诱导巨噬细胞浸润的趋化作用并促进其向替代激活表型（M2）极化，从而增强抗PD1耐药性，进而营造出免疫抑制微环境。从机制上讲，MORF4L2作为NuA4组蛋白乙酰转移酶（HAT）复合物的一部分，有助于组蛋白4赖氨酸-12乙酰化（H4K12Ac）并激活巨噬细胞集落刺激因子（CSF1）的下游转录。CSF1由肿瘤细胞分泌并与巨噬细胞表面的CSF1受体（CSF1R）结合，将巨噬细胞趋化转化并极化为M2表型。此外，我们发现颗粒头样转录因子2（GRHL2）可通过与MORF4L2增强子区域结合来促进MORF4L2转录。值得注意的是，CSF1R抑制剂BLZ549通过阻断GRHL2/MORF4L2/H4K12Ac/CSF1轴恢复了抗PD1敏感性。