Radiotherapy‐resistant prostate cancer cells escape immune checkpoint blockade through the senescence‐related ataxia telangiectasia and Rad3‐related protein

Abstract Background The majority of patients with prostate cancer (PCa) exhibit intrinsic resistance to immune checkpoint blockade (ICB) following radiotherapy (RT). This resistance is generally attributed to the limited antigen presentation of heterogeneous cells within tumors. Here, we aimed to is...

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Main Authors: Chenyi Shao, Yingyi Zhang, Hang Li, Jiajia Chen, Ting Huang, Jiaze Li, Simeng Wen, Sen Wang, Saijun Fan, Yu Zhao
Format: Article
Language:English
Published: Wiley 2025-03-01
Series:Cancer Communications
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Online Access:https://doi.org/10.1002/cac2.12636
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author Chenyi Shao
Yingyi Zhang
Hang Li
Jiajia Chen
Ting Huang
Jiaze Li
Simeng Wen
Sen Wang
Saijun Fan
Yu Zhao
author_facet Chenyi Shao
Yingyi Zhang
Hang Li
Jiajia Chen
Ting Huang
Jiaze Li
Simeng Wen
Sen Wang
Saijun Fan
Yu Zhao
author_sort Chenyi Shao
collection DOAJ
description Abstract Background The majority of patients with prostate cancer (PCa) exhibit intrinsic resistance to immune checkpoint blockade (ICB) following radiotherapy (RT). This resistance is generally attributed to the limited antigen presentation of heterogeneous cells within tumors. Here, we aimed to isolate and characterize these diverse subgroups of tumor post‐RT to understand the molecular mechanisms of their resistance to ICB. Methods Single‐cell RNA‐sequencing (scRNA‐seq) was used to profile senescent cancer cell clusters induced by RT in LNCaP cells. The expression and phosphorylation levels of ataxia telangiectasia and Rad3‐related protein (ATR) were assessed by immunohistochemistry in clinical samples from patients with or without RT. Co‐immunoprecipitation, mutagenesis, and Western blotting were used to measure the interactions between proteins. Xenograft experiments were performed to assess the tumor immune response in the mice. Results We identified a subset of PCa cells that exhibited resistance to RT, characterized by a reduced antigen presentation capability, which enhanced their ability to evade immune detection and resist cytotoxic T‐lymphocyte‐associated protein 4 (CTLA‐4) blockade. scRNA‐seq revealed that the senescent state was a transient phase of PCa cells post‐RT, particularly in CTLA‐4 blockade treatment‐resistant cells. This state was marked by increased cytosolic ATR level. Cytosolic ATR phosphorylated CD86 in its cytosolic domain and enhanced the interaction between CD86 and its E3 ligase MARCH1 through electrostatic attraction. Depletion or inhibition of Atr increased the sensitivity to immune attack and improved responses to anti‐Ctla‐4 antibody treatment in a mouse model. Conclusions Our findings indicate that the activation of cytosolic ATR, which is associated with cellular senescence, impedes the effectiveness of combined RT and ICB treatments. This discovery may provide valuable insights for improving the efficacy of combined RT and ICB therapies in PCa.
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spelling doaj-art-ad58dae963e8486c8e6d9d99edc91c0e2025-08-20T02:10:46ZengWileyCancer Communications2523-35482025-03-0145321824410.1002/cac2.12636Radiotherapy‐resistant prostate cancer cells escape immune checkpoint blockade through the senescence‐related ataxia telangiectasia and Rad3‐related proteinChenyi Shao0Yingyi Zhang1Hang Li2Jiajia Chen3Ting Huang4Jiaze Li5Simeng Wen6Sen Wang7Saijun Fan8Yu Zhao9Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine Institute of Radiation Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Tianjin P. R. ChinaSchool of Disaster and Emergency Medicine Tianjin University Tianjin P. R. ChinaTianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine Institute of Radiation Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Tianjin P. R. ChinaDepartment of Oncology Shengjing Hospital of China Medical University Shenyang Liaoning P. R. ChinaTianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine Institute of Radiation Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Tianjin P. R. ChinaTianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine Institute of Radiation Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Tianjin P. R. ChinaDepartment of Urology The Second Hospital of Tianjin Medical University Tianjin Medical University Tianjin P. R. ChinaTianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine Institute of Radiation Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Tianjin P. R. ChinaInstitute of Radiation Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Tianjin P. R. ChinaTianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine Institute of Radiation Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Tianjin P. R. ChinaAbstract Background The majority of patients with prostate cancer (PCa) exhibit intrinsic resistance to immune checkpoint blockade (ICB) following radiotherapy (RT). This resistance is generally attributed to the limited antigen presentation of heterogeneous cells within tumors. Here, we aimed to isolate and characterize these diverse subgroups of tumor post‐RT to understand the molecular mechanisms of their resistance to ICB. Methods Single‐cell RNA‐sequencing (scRNA‐seq) was used to profile senescent cancer cell clusters induced by RT in LNCaP cells. The expression and phosphorylation levels of ataxia telangiectasia and Rad3‐related protein (ATR) were assessed by immunohistochemistry in clinical samples from patients with or without RT. Co‐immunoprecipitation, mutagenesis, and Western blotting were used to measure the interactions between proteins. Xenograft experiments were performed to assess the tumor immune response in the mice. Results We identified a subset of PCa cells that exhibited resistance to RT, characterized by a reduced antigen presentation capability, which enhanced their ability to evade immune detection and resist cytotoxic T‐lymphocyte‐associated protein 4 (CTLA‐4) blockade. scRNA‐seq revealed that the senescent state was a transient phase of PCa cells post‐RT, particularly in CTLA‐4 blockade treatment‐resistant cells. This state was marked by increased cytosolic ATR level. Cytosolic ATR phosphorylated CD86 in its cytosolic domain and enhanced the interaction between CD86 and its E3 ligase MARCH1 through electrostatic attraction. Depletion or inhibition of Atr increased the sensitivity to immune attack and improved responses to anti‐Ctla‐4 antibody treatment in a mouse model. Conclusions Our findings indicate that the activation of cytosolic ATR, which is associated with cellular senescence, impedes the effectiveness of combined RT and ICB treatments. This discovery may provide valuable insights for improving the efficacy of combined RT and ICB therapies in PCa.https://doi.org/10.1002/cac2.12636ATRCD86immune checkpointsenescence
spellingShingle Chenyi Shao
Yingyi Zhang
Hang Li
Jiajia Chen
Ting Huang
Jiaze Li
Simeng Wen
Sen Wang
Saijun Fan
Yu Zhao
Radiotherapy‐resistant prostate cancer cells escape immune checkpoint blockade through the senescence‐related ataxia telangiectasia and Rad3‐related protein
Cancer Communications
ATR
CD86
immune checkpoint
senescence
title Radiotherapy‐resistant prostate cancer cells escape immune checkpoint blockade through the senescence‐related ataxia telangiectasia and Rad3‐related protein
title_full Radiotherapy‐resistant prostate cancer cells escape immune checkpoint blockade through the senescence‐related ataxia telangiectasia and Rad3‐related protein
title_fullStr Radiotherapy‐resistant prostate cancer cells escape immune checkpoint blockade through the senescence‐related ataxia telangiectasia and Rad3‐related protein
title_full_unstemmed Radiotherapy‐resistant prostate cancer cells escape immune checkpoint blockade through the senescence‐related ataxia telangiectasia and Rad3‐related protein
title_short Radiotherapy‐resistant prostate cancer cells escape immune checkpoint blockade through the senescence‐related ataxia telangiectasia and Rad3‐related protein
title_sort radiotherapy resistant prostate cancer cells escape immune checkpoint blockade through the senescence related ataxia telangiectasia and rad3 related protein
topic ATR
CD86
immune checkpoint
senescence
url https://doi.org/10.1002/cac2.12636
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