Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancer

Abstract Despite advances in systemic therapeutic approaches, metastatic colorectal cancer (mCRC) patients harboring BRAF or RAS mutations have poor outcomes. Cancer stem cells (CSCs) play central roles in drug resistance and CRC recurrence. Therefore, targeting the epigenetic mechanisms that sustai...

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Main Authors: Maria Serena Roca, Rita Lombardi, Cristina Testa, Federica Iannelli, Laura Grumetti, Tania Moccia, Veronica Barile, Laura Addi, Domenico Memoli, Alessandra Leone, Simone Di Franco, Giorgio Stassi, Antonio Avallone, Francesca Bruzzese, Biagio Pucci, Alfredo Budillon, Elena Di Gennaro
Format: Article
Language:English
Published: Nature Publishing Group 2025-08-01
Series:Cell Death and Disease
Online Access:https://doi.org/10.1038/s41419-025-07902-8
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author Maria Serena Roca
Rita Lombardi
Cristina Testa
Federica Iannelli
Laura Grumetti
Tania Moccia
Veronica Barile
Laura Addi
Domenico Memoli
Alessandra Leone
Simone Di Franco
Giorgio Stassi
Antonio Avallone
Francesca Bruzzese
Biagio Pucci
Alfredo Budillon
Elena Di Gennaro
author_facet Maria Serena Roca
Rita Lombardi
Cristina Testa
Federica Iannelli
Laura Grumetti
Tania Moccia
Veronica Barile
Laura Addi
Domenico Memoli
Alessandra Leone
Simone Di Franco
Giorgio Stassi
Antonio Avallone
Francesca Bruzzese
Biagio Pucci
Alfredo Budillon
Elena Di Gennaro
author_sort Maria Serena Roca
collection DOAJ
description Abstract Despite advances in systemic therapeutic approaches, metastatic colorectal cancer (mCRC) patients harboring BRAF or RAS mutations have poor outcomes. Cancer stem cells (CSCs) play central roles in drug resistance and CRC recurrence. Therefore, targeting the epigenetic mechanisms that sustain CSC properties is a promising therapeutic approach. In this study, we report the efficacy of a treatment strategy with the potential to overcome chemotherapy resistance that involves administering the well-known antiepileptic drug and epigenetic agent valproic acid (VPA) and the standard chemotherapy regimen of oxaliplatin/fluoropyrimidine to wild-type CSCs and CSCs with BRAF and RAS mutations in enriched primary spheroid cultures. Notably, we demonstrated that VPA plus chemotherapy was more effective than other epigenetic drug-chemotherapy combinations by inhibiting cell proliferation and clonogenic growth and by inducing apoptosis and DNA damage. Mechanistically, proteomic analysis demonstrated that VPA induced CSC differentiation through the critical target of VPA, β-Catenin. Indeed, VPA promoted the proteasome-dependent degradation of β-Catenin by enhancing its binding to the E2 ubiquitin-conjugating enzyme UBE2a, leading to marked reductions in nuclear and cytoplasmic β-Catenin levels and subsequently decreasing β-Catenin/TCF-LEF target promoter activation. These effects were confirmed in three in vivo CRC xenograft models, including a syngeneic CT26 immunocompetent mouse model, where VPA combined with oxaliplatin/capecitabine chemotherapy and anti-VEGF therapy, a standard first-line treatment for mCRC, significantly suppressed tumor growth and prolonged survival with minimal toxicity. Proteomic analysis of tumor tissues from in vivo CRC models confirmed the VPA-mediated downregulation of CSC markers and β-Catenin.
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spelling doaj-art-76804756af9a49e99003a697fc7dbd2e2025-08-20T03:46:24ZengNature Publishing GroupCell Death and Disease2041-48892025-08-0116111410.1038/s41419-025-07902-8Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancerMaria Serena Roca0Rita Lombardi1Cristina Testa2Federica Iannelli3Laura Grumetti4Tania Moccia5Veronica Barile6Laura Addi7Domenico Memoli8Alessandra Leone9Simone Di Franco10Giorgio Stassi11Antonio Avallone12Francesca Bruzzese13Biagio Pucci14Alfredo Budillon15Elena Di Gennaro16Experimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSExperimental Animal Unit -Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSDepartment of Medicine, Surgery and Dentistry ‘Scuola Medica Salernitana’, University of SalernoExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSDepartment of Precision Medicine in Medical, Surgical and Critical Care, University of PalermoDepartment of Precision Medicine in Medical, Surgical and Critical Care, University of PalermoAbdomen Medical Oncology Unit, Istituto Nazionale Tumori-IRCCS- Fondazione G. PascaleExperimental Animal Unit -Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSScientific Directorate, Istituto Nazionale Tumori-IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori Fondazione G. Pascale - IRCCSAbstract Despite advances in systemic therapeutic approaches, metastatic colorectal cancer (mCRC) patients harboring BRAF or RAS mutations have poor outcomes. Cancer stem cells (CSCs) play central roles in drug resistance and CRC recurrence. Therefore, targeting the epigenetic mechanisms that sustain CSC properties is a promising therapeutic approach. In this study, we report the efficacy of a treatment strategy with the potential to overcome chemotherapy resistance that involves administering the well-known antiepileptic drug and epigenetic agent valproic acid (VPA) and the standard chemotherapy regimen of oxaliplatin/fluoropyrimidine to wild-type CSCs and CSCs with BRAF and RAS mutations in enriched primary spheroid cultures. Notably, we demonstrated that VPA plus chemotherapy was more effective than other epigenetic drug-chemotherapy combinations by inhibiting cell proliferation and clonogenic growth and by inducing apoptosis and DNA damage. Mechanistically, proteomic analysis demonstrated that VPA induced CSC differentiation through the critical target of VPA, β-Catenin. Indeed, VPA promoted the proteasome-dependent degradation of β-Catenin by enhancing its binding to the E2 ubiquitin-conjugating enzyme UBE2a, leading to marked reductions in nuclear and cytoplasmic β-Catenin levels and subsequently decreasing β-Catenin/TCF-LEF target promoter activation. These effects were confirmed in three in vivo CRC xenograft models, including a syngeneic CT26 immunocompetent mouse model, where VPA combined with oxaliplatin/capecitabine chemotherapy and anti-VEGF therapy, a standard first-line treatment for mCRC, significantly suppressed tumor growth and prolonged survival with minimal toxicity. Proteomic analysis of tumor tissues from in vivo CRC models confirmed the VPA-mediated downregulation of CSC markers and β-Catenin.https://doi.org/10.1038/s41419-025-07902-8
spellingShingle Maria Serena Roca
Rita Lombardi
Cristina Testa
Federica Iannelli
Laura Grumetti
Tania Moccia
Veronica Barile
Laura Addi
Domenico Memoli
Alessandra Leone
Simone Di Franco
Giorgio Stassi
Antonio Avallone
Francesca Bruzzese
Biagio Pucci
Alfredo Budillon
Elena Di Gennaro
Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancer
Cell Death and Disease
title Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancer
title_full Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancer
title_fullStr Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancer
title_full_unstemmed Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancer
title_short Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancer
title_sort valproic acid improves the efficacy of oxaliplatin fluoropyrimidine based chemotherapy by targeting cancer stem cell via β catenin modulation in colorectal cancer
url https://doi.org/10.1038/s41419-025-07902-8
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