Single-cell RNA-sequencing atlas reveals an MDK-dependent immunosuppressive environment in ErbB pathway-mutated gallbladder cancer
Single-cell RNA-sequencing atlas reveals an MDK-dependent immunosuppressive environment in ErbB pathway-mutated gallbladder cancer
复制标题
单细胞 RNA 测序图谱揭示了 ErbB 通路突变胆囊癌中 MDK 依赖性免疫抑制环境。
DOI：
10.1016/j.jhep.2021.06.023
复制
发表时间：
2021-10-15
影响因子：
25.7
通讯作者：
Liu, Yingbin
中科院分区：
文献类型：
--
作者：
Zhang, Yijian;Zuo, Chunman;Liu, Yingbin
背景和目标：我们先前的基因组全外显子组测序（WES）数据确定了在调节胆囊癌（GBC）恶性程度中起重要作用的关键ErbB通路突变。在这里，我们测试的假设，个别细胞成分的肿瘤微环境（TME）在GBC功能差异参与ErbB通路突变依赖的肿瘤progress.Methods：我们从事单细胞RNA测序，以揭示转录组异质性和细胞间串扰从13人GBC和邻近正常组织。此外，我们进行了WES分析，以揭示与肿瘤恶性相关的基因组变异。各种批量RNA测序，免疫组化染色，免疫荧光染色和功能实验，研究组织之间的差异有或无ErbB通路mutations.Results：我们确定了16种细胞类型，从总共114,927细胞，其中上皮细胞，M2巨噬细胞，调节性T细胞在ErbB通路突变的肿瘤中占主导地位。腺癌中以1、2、3型上皮细胞为主，腺鳞癌中以4型上皮细胞为主。与ErbB途径突变的肿瘤相比，具有ErbB途径突变的肿瘤具有更大的上皮细胞亚型1和2的群体，并且表达更高水平的分泌性中期因子（MDK）。MDK增加导致与其受体LRP 1的相互作用，该受体由肿瘤浸润性巨噬细胞表达，并促进免疫抑制性巨噬细胞分化。此外，巨噬细胞分泌的CXCL 10和调节性T细胞上的CXCR 3之间的串扰在具有ErbB通路突变的GBC中被诱导。MDK升高与GBC.Conclusions患者的总体生存率差相关：本研究提供了有价值的见解转录组异质性和全球细胞网络的TME，协调功能，以促进GBC的进展与ErbB通路突变，从而揭示新的细胞和分子靶点的癌症治疗。敷设总结：我们采用单细胞RNA测序和功能测定来揭示胆囊癌中存在的转录组异质性和细胞间串扰。我们发现ErbB通路突变降低了抗癌免疫力并导致癌症发展。ErbB途径突变导致免疫抑制性巨噬细胞分化和调节性T细胞活化，解释了在具有这些突变的患者中观察到的抗癌免疫力降低和总生存率降低。(C)2021年，任作者。由Elsevier B. V.代表欧洲肝脏研究协会出版。
Background & Aims: Our previous genomic whole-exome sequencing (WES) data identified the key ErbB pathway mutations that play an essential role in regulating the malignancy of gallbladder cancer (GBC). Herein, we tested the hypothesis that individual cellular components of the tumor microenvironment (TME) in GBC function differentially to participate in ErbB pathway mutation-dependent tumor progression.Methods: We engaged single-cell RNA-sequencing to reveal transcriptomic heterogeneity and intercellular crosstalk from 13 human GBCs and adjacent normal tissues. In addition, we performed WES analysis to reveal the genomic variations related to tumor malignancy. A variety of bulk RNA-sequencing, immunohistochemical staining, immunofluorescence staining and functional experiments were employed to study the difference between tissues with or without ErbB pathway mutations.Results: We identified 16 cell types from a total of 114,927 cells, in which epithelial cells, M2 macrophages, and regulatory T cells were predominant in tumors with ErbB pathway mutations. Furthermore, epithelial cell subtype 1, 2 and 3 were mainly found in adenocarcinoma and subtype 4 was present in adenosquamous carcinoma. The tumors with ErbB pathway mutations harbored larger populations of epithelial cell subtype 1 and 2, and expressed higher levels of secreted midkine (MDK) than tumors without ErbB pathway mutations. Increased MDK resulted in an interaction with its receptor LRP1, which is expressed by tumor-infiltrating macrophages, and promoted immunosuppressive macrophage differentiation. Moreover, the crosstalk between macrophage-secreted CXCL10 and its receptor CXCR3 on regulatory T cells was induced in GBC with ErbB pathway mutations. Elevated MDK was correlated with poor overall survival in patients with GBC.Conclusions: This study has provided valuable insights into transcriptomic heterogeneity and the global cellular network in the TME, which coordinately functions to promote the progres-sion of GBC with ErbB pathway mutations; thus, unveiling novel cellular and molecular targets for cancer therapy. Lay summary: We employed single-cell RNA-sequencing and functional assays to uncover the transcriptomic heterogeneity and intercellular crosstalk present in gallbladder cancer. We found that ErbB pathway mutations reduced anti-cancer im-munity and led to cancer development. ErbB pathway mutations resulted in immunosuppressive macrophage differentiation and regulatory T cell activation, explaining the reduced anti-cancer immunity and worse overall survival observed in patients with these mutations. (C) 2021 The Author(s). Published by Elsevier B.V. on behalf of European Association for the Study of the Liver.