Malevolent alliance of MYBL2hi cancer stem cell and SPP1+ macrophage confers resistance to neoadjuvant immunotherapy in bladder cancer

Background Neoadjuvant immune checkpoint blockade (nICB) has revolutionized cancer treatment, yet the underlying mechanisms of resistance in bladder cancer remain to be explored.Methods We conducted single-cell RNA sequencing (scRNA-seq) on peripheral blood mononuclear cells, tumor tissues, adjacent...

Full description

Saved in:
Bibliographic Details
Main Authors: Lin Ma, Jie Dong, Yingjie Li, Xuebin Zhang, Ziyi Li, Hualin Chen, Yi Xie, Zhaoheng Jin, Yueqiang Peng, Zhigang Ji
Format: Article
Language:English
Published: BMJ Publishing Group 2025-07-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/13/7/e011319.full
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849428303006400512
author Lin Ma
Jie Dong
Yingjie Li
Xuebin Zhang
Ziyi Li
Hualin Chen
Yi Xie
Zhaoheng Jin
Yueqiang Peng
Zhigang Ji
author_facet Lin Ma
Jie Dong
Yingjie Li
Xuebin Zhang
Ziyi Li
Hualin Chen
Yi Xie
Zhaoheng Jin
Yueqiang Peng
Zhigang Ji
author_sort Lin Ma
collection DOAJ
description Background Neoadjuvant immune checkpoint blockade (nICB) has revolutionized cancer treatment, yet the underlying mechanisms of resistance in bladder cancer remain to be explored.Methods We conducted single-cell RNA sequencing (scRNA-seq) on peripheral blood mononuclear cells, tumor tissues, adjacent normal tissues, and metastatic lymph nodes from 2 nICB-naïve and 10 nICB-treated patients with bladder cancer (5 responders and 5 non-responders). Spatial RNA sequencing was performed on tumor slides from two responders and four non-responders. Findings were validated by multiplex immunohistochemistry, mice orthotopic bladder cancer model, and flow cytometry assays.Results nICB remodeled the tumor microenvironment of bladder cancer from both single-cell and spatial perspectives. scRNA-seq analysis revealed a significant increase in MYBL2hi cancer stem cells (CSCs) among non-responders. Analysis of the myeloid population showed that SPP1+ macrophages associated with angiogenesis were linked to CD8+ T cell exclusion. Further investigation into cell–cell communication revealed a propensity for bidirectional crosstalk between MYBL2hi CSCs and SPP1+ macrophages in non-responders. MYBL2hi CSCs derived CCL15, which bound to CCR1 and induced SPP1 upregulation in macrophages which reciprocally enhanced bladder cancer stemness and resistance to nICB through the SPP1-ITGα9β1 axis. Additionally, we identified an aged CCL3+ neutrophil population that interacted with SPP1+ macrophages through a positive feedback loop, contributing to nICB resistance. Finally, in vivo studies demonstrated that combined MYBL2 knockdown and SPP1 targeting synergistically enhanced ICB efficacy in bladder cancer.Conclusions Our research reveals transcriptomic characteristics associated with distinct therapeutic responses to nICB treatment, offering a foundation for optimizing personalized neoadjuvant strategies in bladder cancer.
format Article
id doaj-art-a33c62e726b6451c857b49e078ae7367
institution Kabale University
issn 2051-1426
language English
publishDate 2025-07-01
publisher BMJ Publishing Group
record_format Article
series Journal for ImmunoTherapy of Cancer
spelling doaj-art-a33c62e726b6451c857b49e078ae73672025-08-20T03:28:44ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262025-07-0113710.1136/jitc-2024-011319Malevolent alliance of MYBL2hi cancer stem cell and SPP1+ macrophage confers resistance to neoadjuvant immunotherapy in bladder cancerLin Ma0Jie Dong1Yingjie Li2Xuebin Zhang3Ziyi Li4Hualin Chen5Yi Xie6Zhaoheng Jin7Yueqiang Peng8Zhigang Ji91 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, China1 Department of Urology, Peking Union Medical College Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), Beijing, ChinaBackground Neoadjuvant immune checkpoint blockade (nICB) has revolutionized cancer treatment, yet the underlying mechanisms of resistance in bladder cancer remain to be explored.Methods We conducted single-cell RNA sequencing (scRNA-seq) on peripheral blood mononuclear cells, tumor tissues, adjacent normal tissues, and metastatic lymph nodes from 2 nICB-naïve and 10 nICB-treated patients with bladder cancer (5 responders and 5 non-responders). Spatial RNA sequencing was performed on tumor slides from two responders and four non-responders. Findings were validated by multiplex immunohistochemistry, mice orthotopic bladder cancer model, and flow cytometry assays.Results nICB remodeled the tumor microenvironment of bladder cancer from both single-cell and spatial perspectives. scRNA-seq analysis revealed a significant increase in MYBL2hi cancer stem cells (CSCs) among non-responders. Analysis of the myeloid population showed that SPP1+ macrophages associated with angiogenesis were linked to CD8+ T cell exclusion. Further investigation into cell–cell communication revealed a propensity for bidirectional crosstalk between MYBL2hi CSCs and SPP1+ macrophages in non-responders. MYBL2hi CSCs derived CCL15, which bound to CCR1 and induced SPP1 upregulation in macrophages which reciprocally enhanced bladder cancer stemness and resistance to nICB through the SPP1-ITGα9β1 axis. Additionally, we identified an aged CCL3+ neutrophil population that interacted with SPP1+ macrophages through a positive feedback loop, contributing to nICB resistance. Finally, in vivo studies demonstrated that combined MYBL2 knockdown and SPP1 targeting synergistically enhanced ICB efficacy in bladder cancer.Conclusions Our research reveals transcriptomic characteristics associated with distinct therapeutic responses to nICB treatment, offering a foundation for optimizing personalized neoadjuvant strategies in bladder cancer.https://jitc.bmj.com/content/13/7/e011319.full
spellingShingle Lin Ma
Jie Dong
Yingjie Li
Xuebin Zhang
Ziyi Li
Hualin Chen
Yi Xie
Zhaoheng Jin
Yueqiang Peng
Zhigang Ji
Malevolent alliance of MYBL2hi cancer stem cell and SPP1+ macrophage confers resistance to neoadjuvant immunotherapy in bladder cancer
Journal for ImmunoTherapy of Cancer
title Malevolent alliance of MYBL2hi cancer stem cell and SPP1+ macrophage confers resistance to neoadjuvant immunotherapy in bladder cancer
title_full Malevolent alliance of MYBL2hi cancer stem cell and SPP1+ macrophage confers resistance to neoadjuvant immunotherapy in bladder cancer
title_fullStr Malevolent alliance of MYBL2hi cancer stem cell and SPP1+ macrophage confers resistance to neoadjuvant immunotherapy in bladder cancer
title_full_unstemmed Malevolent alliance of MYBL2hi cancer stem cell and SPP1+ macrophage confers resistance to neoadjuvant immunotherapy in bladder cancer
title_short Malevolent alliance of MYBL2hi cancer stem cell and SPP1+ macrophage confers resistance to neoadjuvant immunotherapy in bladder cancer
title_sort malevolent alliance of mybl2hi cancer stem cell and spp1 macrophage confers resistance to neoadjuvant immunotherapy in bladder cancer
url https://jitc.bmj.com/content/13/7/e011319.full
work_keys_str_mv AT linma malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT jiedong malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT yingjieli malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT xuebinzhang malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT ziyili malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT hualinchen malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT yixie malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT zhaohengjin malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT yueqiangpeng malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer
AT zhigangji malevolentallianceofmybl2hicancerstemcellandspp1macrophageconfersresistancetoneoadjuvantimmunotherapyinbladdercancer