上海交通大学学报（医学版）

上海交通大学学报（医学版） >

2022 , Vol. 42 >Issue 4: 472 - 481

DOI: https://doi.org/10.3969/j.issn.1674-8115.2022.04.009

论著 · 基础研究

多组学数据解析循环及浸润性B细胞在结直肠癌免疫微环境中的作用

  • 龚其雨 ,
  • 陈磊
展开
  • 上海交通大学医学院上海市免疫学研究所，上海 200025
龚其雨（1997—），女，硕士生；电子信箱：gongqiyu@sjtu.edu.cn。
陈磊，电子信箱：lei.chen@sjtu.edu.cn。

收稿日期: 2021-11-18

  录用日期: 2022-03-28

  网络出版日期: 2022-04-28

收起

Role of circulating and infiltrating B cells in immune microenvironment of colorectal cancer by multi-omics data profiling

  • Qiyu GONG ,
  • Lei CHEN
Expand
  • Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
CHEN Lei, E-mail: lei.chen@sjtu.edu.cn.

Received date: 2021-11-18

  Accepted date: 2022-03-28

  Online published: 2022-04-28

Fold

摘要

目的·探究循环及浸润性B细胞亚群在结直肠癌免疫微环境中的作用，探寻B细胞发育的潜在驱动机制，分析免疫微环境中B细胞与其他免疫细胞的互作关系及通信分子的功能，并寻找可能的调控途径以促进B细胞的抗肿瘤效应。方法·收集3例结直肠癌患者的癌旁（正常）、肿瘤和外周血共计9份样本，分别制备单细胞悬液。通过多色流式细胞仪分选免疫细胞后，利用10X Genomics平台进行单细胞转录组和B细胞受体组库（B cell receptor repertoire，BCR）测序，并使用Seurat等生信工具进行数据分析，从肿瘤与正常组织中B细胞的浸润数量及功能方面的差异、B细胞亚群与T细胞以及与髓系细胞亚群的互作关系、B细胞亚群的转录因子调控网络等角度进行探索分析。结果·与正常组织相比较，CD20+B细胞亚群在肿瘤组织中的浸润数量显著上升；其中，以生发中心B细胞（germinal center B cells，GCB）的作用最突出，发生积极的克隆扩增和重链突变水平的提升，并且向记忆B细胞分化的趋势增加。然而，浆细胞在肿瘤微环境中的数量大幅减少，以分泌IgA类抗体的浆细胞减少最为明显。此外，相较正常组织的免疫微环境，肿瘤组织中的GCB细胞与T细胞等其他免疫细胞的联系变得更加紧密，正向调控免疫功能的通信分子对显著富集。结论·GCB在结直肠癌肿瘤微环境中的作用大幅提升，以自身显著提升的重链突变水平来提高其对肿瘤抗原的亲和力，同时和微环境中的免疫细胞联系增强，发挥着积极的抗肿瘤效应。

关键词： 生物信息学; 单细胞测序; 结直肠癌; 免疫学; B细胞

本文引用格式

龚其雨 , 陈磊 . 多组学数据解析循环及浸润性B细胞在结直肠癌免疫微环境中的作用[J]. 上海交通大学学报（医学版）, 2022 , 42(4) : 472 -481 . DOI: 10.3969/j.issn.1674-8115.2022.04.009

Abstract

Objective

·To explore the role of circulating and infiltrating B cell subsets in the immune microenvironment of colorectal cancer and discover the potential molecular-driving mechanisms behind the above transformation. Then to analyze the interaction patterns between B cell subsets and other immune cells in the microenvironment, including the function analysis of communicating molecules. All these steps assist to find possible regulatory ways to augment the anti-tumor effect of B cells.

Methods

·Paired tumor, normal and peripheral blood tissues were collected from three patients with colorectal cancer. A total of 9 samples were applied to prepare the single-cell suspension. Then all immune cells were sorted by flow cytometry. 10X Genomics platform was applied for single-cell transcriptome and B cell receptor repertoire (BCR) sequencing. Bioinformatic tools such as Seurat etc. were taken to analyze scRNA and BCR data. The downstream analysis includes differential analysis of B cell in tumor versus normal tissues, cell interaction analysis in the immune microenvironment and regulatory networks of B cell subsets.

Results

·CD20+B cells were significantly increased in tumor versus normal tissues, especially germinal center B cells (GCB), which showed positively clonal amplification and high mutation level. Besides, GCB showed the tendency of transformation into memory B cells. On the contrary, plasma cells significantly reduced their activity in tumor, which was dominated by the IgA class. In addition, comparing with the immune microenvironment in normal tissues, GCB in tumor tissues are more closely connected with T cells and other immune cells, and the communication molecule pairs that positively regulate immune function were significantly enriched.

Conclusion

·The importance of GCB in the immune microenvironment of colorectal cancer is greatly increased. And they exert a positive anti-tumor effect through the up-regulated heavy-chain mutation level and their close association with surrounding immune cells.

Key words： bioinformatics; single cell sequencing; colorectal cancer; immunology; B cell

参考文献

1 XIE Y H, CHEN Y X, FANG J Y. Comprehensive review of targeted therapy for colorectal cancer[J]. Signal Transduct Target Ther, 2020, 5(1):22.
2 BINNEWIES M, ROBERTS E W, KERSTEN K, et al. Understanding the tumor immune microenvironment (TIME) for effective therapy[J]. Nat Med, 2018, 24(5):541-550.
3 CIARDIELLO D, VITIELLO P P, CARDONE C, et al. Immunotherapy of colorectal cancer: challenges for therapeutic efficacy[J]. Cancer Treat Rev, 2019, 76: 22-32.
4 ZHAO P, LI L, JIANG X, et al. Mismatch repair deficiency/microsatellite instability-high as a predictor for anti-PD-1/PD-L1 immunotherapy efficacy[J]. J Hematol Oncol, 2019, 12(1): 54.
5 GALON J, COSTES A, SANCHEZ-CABO F, et al. Type, density, and location of immune cells within human colorectal tumors predict clinical outcome[J]. Science, 2006, 313(5795): 1960-1964.
6 GARBER K. Driving T-cell immunotherapy to solid tumors[J]. Nat Biotechnol, 2018, 36(3): 215-219.
7 SHEN P, FILLATREAU S. Antibody-independent functions of B cells: a focus on cytokines[J]. Nat Rev Immunol, 2015, 15(7):441-451.
8 SCHARER C D, PATTERSON D G, MI T, et al. Antibody-secreting cell destiny emerges during the initial stages of B-cell activation[J]. Nat Commun, 2020, 11(1): 3989.
9 GUNDERSON A J, COUSSENS L M. B cells and their mediators as targets for therapy in solid tumors[J]. Exp Cell Res, 2013, 319(11):1644-1649.
10 SHARONOV G V, SEREBROVSKAYA E O, YUZHAKOVA D V, et al. B cells, plasma cells and antibody repertoires in the tumour microenvironment[J]. Nat Rev Immunol, 2020, 20(5): 294-307.
11 BERGOMAS F, GRIZZI F, DONI A, et al. Tertiary intratumor lymphoid tissue in colo-rectal cancer[J]. Cancers (Basel), 2011, 4(1):1-10.
12 SAUTèS-FRIDMAN C, PETITPREZ F, CALDERARO J, et al. Tertiary lymphoid structures in the era of cancer immunotherapy[J]. Nat Rev Cancer, 2019, 19(6): 307-325.
13 SCHULTZE J L, MICHALAK S, SEAMON M J, et al. CD40-activated human B cells: an alternative source of highly efficient antigen presenting cells to generate autologous antigen-specific T cells for adoptive immunotherapy[J]. J Clin Invest, 1997, 100(11):2757-2765.
14 ZHANG W, FENG Q, WANG C, et al. Characterization of the B cell receptor repertoire in the intestinal mucosa and of tumor-infiltrating lymphocytes in colorectal adenoma and carcinoma[J]. J Immunol, 2017, 198(9): 3719-3728.
15 SCHOORL R, RIVIERE A B, BORNE A E, et al. Identification of T and B lymphocytes in human breast cancer with immunohistochemical techniques[J]. Am J Pathol, 1976, 84(3):529-544.
16 POSCH F, SILINA K, LEIBL S, et al. Maturation of tertiary lymphoid structures and recurrence of stage Ⅱ and Ⅲ colorectal cancer[J]. Oncoimmunology, 2018, 7(2): e1378844.
17 ZHANG L, LI Z, SKRZYPCZYNSKA K M, et al. Single-cell analyses inform mechanisms of myeloid-targeted therapies in colon cancer[J]. Cell, 2020, 181(2): 442-459.e29.
18 WANG J, XU Y, CHEN Z, et al. Liver immune profiling reveals pathogenesis and therapeutics for biliary atresia[J]. Cell, 2020, 183(7):1867-1883.e26.
19 KROEGER D R, MILNE K, NELSON B H. Tumor-infiltrating plasma cells are associated with tertiary lymphoid structures, cytolytic T-cell responses, and superior prognosis in ovarian cancer[J]. Clin Cancer Res, 2016, 22(12): 3005-3015.
20 LUND F E, RANDALL T D. Effector and regulatory B cells: modulators of CD4+ T cell immunity[J]. Nat Rev Immunol, 2010, 10(4): 236-247.
21 NIELSEN J S, SAHOTA R A, MILNE K, et al. CD20+ tumor-infiltrating lymphocytes have an atypical CD27- memory phenotype and together with CD8+ T cells promote favorable prognosis in ovarian cancer[J]. Clin Cancer Res, 2012, 18(12): 3281-3292.
22 RIVERA A, CHEN C C, RON N, et al. Role of B cells as antigen-presenting cells in vivo revisited: antigen-specific B cells are essential for T cell expansion in lymph nodes and for systemic T cell responses to low antigen concentrations[J]. Int Immunol, 2001, 13(12): 1583-1593.
Options
文章导航

/

〈 〉