Targeting of the IL-33/Wnt axis restricts breast cancer stemness and metastasis

Abstract Interleukin-33 (IL-33) plays multifaceted roles in tumor progression, but its autocrine regulation of breast cancer stemness and metastasis via the Wnt pathway remains unclear. Here, we investigated the IL-33/ST2 axis in breast cancer using CRISPR/Cas9, single-cell RNA sequencing, and murin...

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Main Authors: Guanglin Fan, Shuting Zuo, Zhen Wang, Siwei Zhang, Liping Liu, Haoge Luo, Yingdong Xie, Yan Zhang, Dong Li
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-03260-9
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author Guanglin Fan
Shuting Zuo
Zhen Wang
Siwei Zhang
Liping Liu
Haoge Luo
Yingdong Xie
Yan Zhang
Dong Li
author_facet Guanglin Fan
Shuting Zuo
Zhen Wang
Siwei Zhang
Liping Liu
Haoge Luo
Yingdong Xie
Yan Zhang
Dong Li
author_sort Guanglin Fan
collection DOAJ
description Abstract Interleukin-33 (IL-33) plays multifaceted roles in tumor progression, but its autocrine regulation of breast cancer stemness and metastasis via the Wnt pathway remains unclear. Here, we investigated the IL-33/ST2 axis in breast cancer using CRISPR/Cas9, single-cell RNA sequencing, and murine models (orthotopic 4T1 and spontaneous MMTV-PyMT). Elevated IL-33 levels correlated with aggressive subtypes and poor prognosis. IL-33 overexpression enhanced proliferation, migration, and cancer stem cell (CSC) marker expression (CD44, ALDH1) in 4T1 and MDA-MB-231 cells, whereas ST2 knockdown via CRISPR or adeno-associated virus (AAV) attenuated tumor growth and metastasis in vivo, reducing CSC frequency. Mechanistically, IL-33 activated Wnt/β-catenin signaling to promote stemness, which was reversed by the Wnt inhibitor XAV-939. Single-cell analysis revealed that IL-33 overexpression skewed the immune microenvironment toward immunosuppression, while ST2 knockdown restored antitumor immunity. Our findings establish an IL-33–Wnt axis as a critical driver of breast cancer aggressiveness and propose AAV-mediated ST2 silencing as a novel therapeutic strategy. Targeting this axis may offer dual benefits by suppressing stemness and enhancing immune surveillance, warranting clinical exploration for advanced breast cancer.
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spelling doaj-art-8d9ae65ee91b4a0a88d83e0cfcfed1852025-08-20T03:08:40ZengNature PortfolioScientific Reports2045-23222025-05-0115111510.1038/s41598-025-03260-9Targeting of the IL-33/Wnt axis restricts breast cancer stemness and metastasisGuanglin Fan0Shuting Zuo1Zhen Wang2Siwei Zhang3Liping Liu4Haoge Luo5Yingdong Xie6Yan Zhang7Dong Li8Department of Breast Surgery, The Second Hospital of Jilin UniversityDepartment of Breast Surgery, The Second Hospital of Jilin UniversityDepartment of Breast Surgery, The Second Hospital of Jilin UniversityDepartment of Pharmacology, College of Basic Medical Sciences, Jilin UniversityDepartment of Immunology, College of Basic Medical Sciences, Jilin UniversityDepartment of Immunology, College of Basic Medical Sciences, Jilin UniversityDepartment of Immunology, College of Basic Medical Sciences, Jilin UniversityDepartment of Breast Surgery, The Second Hospital of Jilin UniversityDepartment of Immunology, College of Basic Medical Sciences, Jilin UniversityAbstract Interleukin-33 (IL-33) plays multifaceted roles in tumor progression, but its autocrine regulation of breast cancer stemness and metastasis via the Wnt pathway remains unclear. Here, we investigated the IL-33/ST2 axis in breast cancer using CRISPR/Cas9, single-cell RNA sequencing, and murine models (orthotopic 4T1 and spontaneous MMTV-PyMT). Elevated IL-33 levels correlated with aggressive subtypes and poor prognosis. IL-33 overexpression enhanced proliferation, migration, and cancer stem cell (CSC) marker expression (CD44, ALDH1) in 4T1 and MDA-MB-231 cells, whereas ST2 knockdown via CRISPR or adeno-associated virus (AAV) attenuated tumor growth and metastasis in vivo, reducing CSC frequency. Mechanistically, IL-33 activated Wnt/β-catenin signaling to promote stemness, which was reversed by the Wnt inhibitor XAV-939. Single-cell analysis revealed that IL-33 overexpression skewed the immune microenvironment toward immunosuppression, while ST2 knockdown restored antitumor immunity. Our findings establish an IL-33–Wnt axis as a critical driver of breast cancer aggressiveness and propose AAV-mediated ST2 silencing as a novel therapeutic strategy. Targeting this axis may offer dual benefits by suppressing stemness and enhancing immune surveillance, warranting clinical exploration for advanced breast cancer.https://doi.org/10.1038/s41598-025-03260-9IL-33ST2 knockdownBreast cancer stem cellsWnt pathwayImmunotherapy
spellingShingle Guanglin Fan
Shuting Zuo
Zhen Wang
Siwei Zhang
Liping Liu
Haoge Luo
Yingdong Xie
Yan Zhang
Dong Li
Targeting of the IL-33/Wnt axis restricts breast cancer stemness and metastasis
Scientific Reports
IL-33
ST2 knockdown
Breast cancer stem cells
Wnt pathway
Immunotherapy
title Targeting of the IL-33/Wnt axis restricts breast cancer stemness and metastasis
title_full Targeting of the IL-33/Wnt axis restricts breast cancer stemness and metastasis
title_fullStr Targeting of the IL-33/Wnt axis restricts breast cancer stemness and metastasis
title_full_unstemmed Targeting of the IL-33/Wnt axis restricts breast cancer stemness and metastasis
title_short Targeting of the IL-33/Wnt axis restricts breast cancer stemness and metastasis
title_sort targeting of the il 33 wnt axis restricts breast cancer stemness and metastasis
topic IL-33
ST2 knockdown
Breast cancer stem cells
Wnt pathway
Immunotherapy
url https://doi.org/10.1038/s41598-025-03260-9
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