Single-cell sequencing reveals antitumor characteristics of intratumoral immune cells in old mice

Background Aging has long been thought to be a major risk factor for various types of cancers. However, accumulating evidence indicates increased resistance of old animals to tumor growth. An in-depth understanding of how old individuals defend against tumor invasion requires further investigations....

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Main Authors: Xin Wang, Lei Lei, Lin Shi, Lianjun Zhang, Xiaobo Zhou, Xiaofeng Yang, Yanhong Su, Cangang Zhang, Kaili Ma, Huiqiang Zheng, Anjun Jiao, Haiyan Liu, Yujing Zou, Chenming Sun, Yuzhu Hou, Zhengtao Xiao, Baojun Zhang
Format: Article
Language:English
Published: BMJ Publishing Group 2021-10-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/9/10/e002809.full
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author Xin Wang
Lei Lei
Lin Shi
Lianjun Zhang
Xiaobo Zhou
Xiaofeng Yang
Yanhong Su
Cangang Zhang
Kaili Ma
Huiqiang Zheng
Anjun Jiao
Haiyan Liu
Yujing Zou
Chenming Sun
Yuzhu Hou
Zhengtao Xiao
Baojun Zhang
author_facet Xin Wang
Lei Lei
Lin Shi
Lianjun Zhang
Xiaobo Zhou
Xiaofeng Yang
Yanhong Su
Cangang Zhang
Kaili Ma
Huiqiang Zheng
Anjun Jiao
Haiyan Liu
Yujing Zou
Chenming Sun
Yuzhu Hou
Zhengtao Xiao
Baojun Zhang
author_sort Xin Wang
collection DOAJ
description Background Aging has long been thought to be a major risk factor for various types of cancers. However, accumulating evidence indicates increased resistance of old animals to tumor growth. An in-depth understanding of how old individuals defend against tumor invasion requires further investigations.Methods We revealed age-associated alterations in tumor-infiltrating immune cells between young and old mice using single-cell RNA and coupled T cell receptor (TCR) sequencing analysis. Multiple bioinformatics methods were adopted to analyze the characteristics of the transcriptome between two groups. To explore the impacts of young and old CD8+ T cells on tumor growth, mice were treated with anti-CD8 antibody every 3 days starting 7 days after tumor inoculation. Flow cytometry was used to validate the differences indicated by sequencing analysis between young and old mice.Results We found a higher proportion of cytotoxic CD8+ T cells, naturally occurring Tregs, conventional dendritic cell (DC), and M1-like macrophages in tumors of old mice compared with a higher percentage of exhausted CD8+ T cells, induced Tregs, plasmacytoid DC, and M2-like macrophages in young mice. Importantly, TCR diversity analysis showed that top 10 TCR clones consisted primarily of exhausted CD8+ T cells in young mice whereas top clones were predominantly cytotoxic CD8+ T cells in old mice. Old mice had more CD8+ T cells with a ‘progenitor’ and less ‘terminally’ exhausted phenotypes than young mice. Consistently, trajectory inference demonstrated that CD8+ T cells preferentially differentiated into cytotoxic cells in old mice in contrast to exhausted cells in young mice. Importantly, elimination of CD8+ T cells in old mice during tumor growth significantly accelerated tumor development. Moreover, senescent features were demonstrated in exhausted but not cytotoxic CD8+ T cells regardless of young and old mice.Conclusions Our data revealed that a significantly higher proportion of effector immune cells in old mice defends against tumor progression, providing insights into understanding the altered kinetics of cancer development and the differential response to immunotherapeutic modulation in elderly patients.
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spelling doaj-art-783c2845ee4449d69fa3de83aaf0ab202025-08-20T01:53:15ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262021-10-0191010.1136/jitc-2021-002809Single-cell sequencing reveals antitumor characteristics of intratumoral immune cells in old miceXin Wang0Lei Lei1Lin Shi2Lianjun Zhang3Xiaobo Zhou4Xiaofeng Yang5Yanhong Su6Cangang Zhang7Kaili Ma8Huiqiang Zheng9Anjun Jiao10Haiyan Liu11Yujing Zou12Chenming Sun13Yuzhu Hou14Zhengtao Xiao15Baojun Zhang16Department of Oncology, Fudan University Shanghai Medical College, Shanghai, People`s Republic of ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaEvidence Based Medicine Center, The Affiliated Traditional Chinese Medicine Hospital, Guangzhou Medical University, Guangzhou, People`s Republic of ChinaJiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute, Xuzhou Medical University, Xuzhou, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaInstitute of Systems Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDivision of Hematologic Malignancies and Cellular Therapy, Duke University Medical Center, Durham, North Carolina, USADepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaDepartment of Pathogenic Microbiology and Immunology, Xi’an Jiaotong University, Xi’an, ChinaBackground Aging has long been thought to be a major risk factor for various types of cancers. However, accumulating evidence indicates increased resistance of old animals to tumor growth. An in-depth understanding of how old individuals defend against tumor invasion requires further investigations.Methods We revealed age-associated alterations in tumor-infiltrating immune cells between young and old mice using single-cell RNA and coupled T cell receptor (TCR) sequencing analysis. Multiple bioinformatics methods were adopted to analyze the characteristics of the transcriptome between two groups. To explore the impacts of young and old CD8+ T cells on tumor growth, mice were treated with anti-CD8 antibody every 3 days starting 7 days after tumor inoculation. Flow cytometry was used to validate the differences indicated by sequencing analysis between young and old mice.Results We found a higher proportion of cytotoxic CD8+ T cells, naturally occurring Tregs, conventional dendritic cell (DC), and M1-like macrophages in tumors of old mice compared with a higher percentage of exhausted CD8+ T cells, induced Tregs, plasmacytoid DC, and M2-like macrophages in young mice. Importantly, TCR diversity analysis showed that top 10 TCR clones consisted primarily of exhausted CD8+ T cells in young mice whereas top clones were predominantly cytotoxic CD8+ T cells in old mice. Old mice had more CD8+ T cells with a ‘progenitor’ and less ‘terminally’ exhausted phenotypes than young mice. Consistently, trajectory inference demonstrated that CD8+ T cells preferentially differentiated into cytotoxic cells in old mice in contrast to exhausted cells in young mice. Importantly, elimination of CD8+ T cells in old mice during tumor growth significantly accelerated tumor development. Moreover, senescent features were demonstrated in exhausted but not cytotoxic CD8+ T cells regardless of young and old mice.Conclusions Our data revealed that a significantly higher proportion of effector immune cells in old mice defends against tumor progression, providing insights into understanding the altered kinetics of cancer development and the differential response to immunotherapeutic modulation in elderly patients.https://jitc.bmj.com/content/9/10/e002809.full
spellingShingle Xin Wang
Lei Lei
Lin Shi
Lianjun Zhang
Xiaobo Zhou
Xiaofeng Yang
Yanhong Su
Cangang Zhang
Kaili Ma
Huiqiang Zheng
Anjun Jiao
Haiyan Liu
Yujing Zou
Chenming Sun
Yuzhu Hou
Zhengtao Xiao
Baojun Zhang
Single-cell sequencing reveals antitumor characteristics of intratumoral immune cells in old mice
Journal for ImmunoTherapy of Cancer
title Single-cell sequencing reveals antitumor characteristics of intratumoral immune cells in old mice
title_full Single-cell sequencing reveals antitumor characteristics of intratumoral immune cells in old mice
title_fullStr Single-cell sequencing reveals antitumor characteristics of intratumoral immune cells in old mice
title_full_unstemmed Single-cell sequencing reveals antitumor characteristics of intratumoral immune cells in old mice
title_short Single-cell sequencing reveals antitumor characteristics of intratumoral immune cells in old mice
title_sort single cell sequencing reveals antitumor characteristics of intratumoral immune cells in old mice
url https://jitc.bmj.com/content/9/10/e002809.full
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