RIPK2 promotes colorectal cancer metastasis by protecting YAP degradation from ITCH-mediated ubiquitination

Abstract Colorectal cancer (CRC) is the second leading cause of cancer-related death worldwide, making the exploration of metastatic mechanisms crucial for therapeutic advancements. In this study, we identified receptor-interacting protein kinase 2 (RIPK2) as an independent risk factor for poor CRC...

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Main Authors: Chen Lu, Hongda Liu, Tianyu Liu, Sizheng Sun, Yanan Zheng, Tao Ling, Xiagang Luo, Yiming E, Yuting Xu, Jie Li, Lei Liu, Lin Miao, Zhengxia Liu, Chunzhao Yu
Format: Article
Language:English
Published: Nature Publishing Group 2025-04-01
Series:Cell Death and Disease
Online Access:https://doi.org/10.1038/s41419-025-07599-9
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author Chen Lu
Hongda Liu
Tianyu Liu
Sizheng Sun
Yanan Zheng
Tao Ling
Xiagang Luo
Yiming E
Yuting Xu
Jie Li
Lei Liu
Lin Miao
Zhengxia Liu
Chunzhao Yu
author_facet Chen Lu
Hongda Liu
Tianyu Liu
Sizheng Sun
Yanan Zheng
Tao Ling
Xiagang Luo
Yiming E
Yuting Xu
Jie Li
Lei Liu
Lin Miao
Zhengxia Liu
Chunzhao Yu
author_sort Chen Lu
collection DOAJ
description Abstract Colorectal cancer (CRC) is the second leading cause of cancer-related death worldwide, making the exploration of metastatic mechanisms crucial for therapeutic advancements. In this study, we identified receptor-interacting protein kinase 2 (RIPK2) as an independent risk factor for poor CRC prognosis. Single-cell RNA sequencing and spatial transcriptomics revealed that a tumor cell cluster with high RIPK2 expression exhibited enhanced metastatic potential, closely linked to bacterial invasion. In vitro and in vivo experiments confirmed that RIPK2 specifically promotes tumor cell migration and invasion, rather than proliferation. Proteomic analysis indicated that RIPK2 knockdown leads to increased proteolysis mediated by ubiquitin, particularly affecting the oncoprotein YAP. Additionally, bacterial invasion of epithelial cells was significantly suppressed in RIPK2 knockdown cells, suggesting a connection to the NOD2-RIPK2 pathway, stimulated by bacterial muramyl dipeptide (MDP). We demonstrated that MDP levels are significantly higher in CRC tissues compared to adjacent non-cancerous tissues, correlating with RIPK2 activation. This activation triggers K63-linked ubiquitination of RIPK2, essential for NF-κB and MAPK pathway activation. Mechanistic studies identified the E3 ubiquitin ligase ITCH as a critical mediator, balancing K63-linked ubiquitination of RIPK2 and K48-linked ubiquitination of YAP, leading to YAP degradation and suppressed CRC metastasis. The stability of YAP could also be disrupted by GSK583, a pharmacological inhibitor of RIPK2, effectively suppressing CRC metastasis. Our findings provide deep insights into RIPK2’s role in CRC progression and present a promising target for future therapeutic strategies.
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spelling doaj-art-dfbf3cbaaaa947ec8baded5469017a852025-08-20T03:08:12ZengNature Publishing GroupCell Death and Disease2041-48892025-04-0116111610.1038/s41419-025-07599-9RIPK2 promotes colorectal cancer metastasis by protecting YAP degradation from ITCH-mediated ubiquitinationChen Lu0Hongda Liu1Tianyu Liu2Sizheng Sun3Yanan Zheng4Tao Ling5Xiagang Luo6Yiming E7Yuting Xu8Jie Li9Lei Liu10Lin Miao11Zhengxia Liu12Chunzhao Yu13Department of General Surgery, Sir Run Run Hospital of Nanjing Medical UniversityDepartment of General Surgery, First Affiliated Hospital of Nanjing Medical UniversityDepartment of General Surgery, Sir Run Run Hospital of Nanjing Medical UniversityDepartment of General Surgery, The Affiliated Taizhou People’s Hospital of Nanjing Medical UniversityDepartment of Geriatrics, The Second Affiliated Hospital of Nanjing Medical UniversityDepartment of Geriatrics, The Second Affiliated Hospital of Nanjing Medical UniversityDepartment of General Surgery, The Second Affiliated Hospital of Nanjing Medical UniversityDepartment of General Surgery, The Second Affiliated Hospital of Nanjing Medical UniversityOphthalmic Oncology Department, Nanjing Medical University Eye HospitalDepartment of Oncology, The Second Affiliated Hospital of Nanjing Medical UniversityDepartment of Gastroenterology, The Affiliated Yixing Hospital of Jiangsu UniversityMedical Centre for Digestive Diseases, The Second Affiliated Hospital of Nanjing Medical UniversityDepartment of Geriatrics, The Second Affiliated Hospital of Nanjing Medical UniversityDepartment of General Surgery, Sir Run Run Hospital of Nanjing Medical UniversityAbstract Colorectal cancer (CRC) is the second leading cause of cancer-related death worldwide, making the exploration of metastatic mechanisms crucial for therapeutic advancements. In this study, we identified receptor-interacting protein kinase 2 (RIPK2) as an independent risk factor for poor CRC prognosis. Single-cell RNA sequencing and spatial transcriptomics revealed that a tumor cell cluster with high RIPK2 expression exhibited enhanced metastatic potential, closely linked to bacterial invasion. In vitro and in vivo experiments confirmed that RIPK2 specifically promotes tumor cell migration and invasion, rather than proliferation. Proteomic analysis indicated that RIPK2 knockdown leads to increased proteolysis mediated by ubiquitin, particularly affecting the oncoprotein YAP. Additionally, bacterial invasion of epithelial cells was significantly suppressed in RIPK2 knockdown cells, suggesting a connection to the NOD2-RIPK2 pathway, stimulated by bacterial muramyl dipeptide (MDP). We demonstrated that MDP levels are significantly higher in CRC tissues compared to adjacent non-cancerous tissues, correlating with RIPK2 activation. This activation triggers K63-linked ubiquitination of RIPK2, essential for NF-κB and MAPK pathway activation. Mechanistic studies identified the E3 ubiquitin ligase ITCH as a critical mediator, balancing K63-linked ubiquitination of RIPK2 and K48-linked ubiquitination of YAP, leading to YAP degradation and suppressed CRC metastasis. The stability of YAP could also be disrupted by GSK583, a pharmacological inhibitor of RIPK2, effectively suppressing CRC metastasis. Our findings provide deep insights into RIPK2’s role in CRC progression and present a promising target for future therapeutic strategies.https://doi.org/10.1038/s41419-025-07599-9
spellingShingle Chen Lu
Hongda Liu
Tianyu Liu
Sizheng Sun
Yanan Zheng
Tao Ling
Xiagang Luo
Yiming E
Yuting Xu
Jie Li
Lei Liu
Lin Miao
Zhengxia Liu
Chunzhao Yu
RIPK2 promotes colorectal cancer metastasis by protecting YAP degradation from ITCH-mediated ubiquitination
Cell Death and Disease
title RIPK2 promotes colorectal cancer metastasis by protecting YAP degradation from ITCH-mediated ubiquitination
title_full RIPK2 promotes colorectal cancer metastasis by protecting YAP degradation from ITCH-mediated ubiquitination
title_fullStr RIPK2 promotes colorectal cancer metastasis by protecting YAP degradation from ITCH-mediated ubiquitination
title_full_unstemmed RIPK2 promotes colorectal cancer metastasis by protecting YAP degradation from ITCH-mediated ubiquitination
title_short RIPK2 promotes colorectal cancer metastasis by protecting YAP degradation from ITCH-mediated ubiquitination
title_sort ripk2 promotes colorectal cancer metastasis by protecting yap degradation from itch mediated ubiquitination
url https://doi.org/10.1038/s41419-025-07599-9
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