Extracellular vesicle-enriched secretome of adipose-derived stem cells upregulates clusterin to alleviate doxorubicin-induced apoptosis in cardiomyocytes

Abstract Doxorubicin (DOX) is a potent chemotherapeutic widely used against various cancers, but its clinical application is limited by DOX-induced cardiotoxicity (DIC). This study explored the cardioprotective potential of extracellular vesicle-enriched secretome derived from adipose stem cells (EV...

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Main Authors: Wan-Tseng Hsu, Shinji Kobuchi, Tung-Chun Russell Chien, I-Chun Chen, Shohei Hamada, Masayuki Tsujimoto, I-Lin Tsai, Yun-Sheng Wong, Kuan-Hsuan Tung, Ying-Zhen He
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Biology Direct
Online Access:https://doi.org/10.1186/s13062-025-00664-5
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author Wan-Tseng Hsu
Shinji Kobuchi
Tung-Chun Russell Chien
I-Chun Chen
Shohei Hamada
Masayuki Tsujimoto
I-Lin Tsai
Yun-Sheng Wong
Kuan-Hsuan Tung
Ying-Zhen He
author_facet Wan-Tseng Hsu
Shinji Kobuchi
Tung-Chun Russell Chien
I-Chun Chen
Shohei Hamada
Masayuki Tsujimoto
I-Lin Tsai
Yun-Sheng Wong
Kuan-Hsuan Tung
Ying-Zhen He
author_sort Wan-Tseng Hsu
collection DOAJ
description Abstract Doxorubicin (DOX) is a potent chemotherapeutic widely used against various cancers, but its clinical application is limited by DOX-induced cardiotoxicity (DIC). This study explored the cardioprotective potential of extracellular vesicle-enriched secretome derived from adipose stem cells (EVSASC) in mitigating DOX-induced apoptosis in cardiomyocytes. Adipose-derived stem cells were cultured, and their conditioned medium and extraceullular vesicles were isolated and characterized according to the Minimal Information for Studies of Extracellular Vesicles 2023 guidelines. HL-1 cardiomyocytes were pretreated with EVSASC before exposure to 1 µM DOX. Cell viability was assessed via the cell counting kit-8 assay, while apoptosis markers and survival mediators were evaluated through Western blotting. RNA sequencing identified differentially expressed genes, including clusterin (Clu), which was further quantified using an enzyme-linked immunosorbent assay. The functional role of clusterin was validated through siRNA-mediated knockdown. EVSASC significantly improved cell viability in DOX-exposed cardiomyocytes and reduced the cleaved caspase-3 to procaspase-3 ratio. Clusterin expression was highest in EVSASC-treated cells, and its knockdown markedly increased caspase-3 cleavage, confirming its pivotal role in cardioprotection. Moreover, EVSASC enhanced the phosphorylation of AKT, Bcl2-associated agonist of cell death, and glycogen synthase kinase-3β, implicating PI3K/AKT pathway activation in clusterin upregulation and anti-apoptotic effects. These findings demonstrate that EVSASC mitigates DOX-induced apoptosis in cardiomyocytes through clusterin upregulation and PI3K/AKT pathway activation. Clusterin is identified as a potential biomarker for evaluating EVSASC efficacy. While EVSASC shows promise as a cardioprotective strategy against DIC, further studies are needed to optimize its therapeutic safety by addressing potential oncogenic risks.
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spelling doaj-art-24cd767ca0e64c818c160ada31792a7e2025-08-20T03:45:56ZengBMCBiology Direct1745-61502025-07-0120112510.1186/s13062-025-00664-5Extracellular vesicle-enriched secretome of adipose-derived stem cells upregulates clusterin to alleviate doxorubicin-induced apoptosis in cardiomyocytesWan-Tseng Hsu0Shinji Kobuchi1Tung-Chun Russell Chien2I-Chun Chen3Shohei Hamada4Masayuki Tsujimoto5I-Lin Tsai6Yun-Sheng Wong7Kuan-Hsuan Tung8Ying-Zhen He9School of Pharmacy, College of Medicine, National Taiwan UniversityLaboratory of Pharmacokinetics, Kyoto Pharmaceutical UniversityGraduate Institute of Clinical Pharmacy, College of Medicine, National Taiwan UniversityDepartment of Medical Oncology, National Taiwan University Cancer CenterDivision of Pharmaceutical Sciences, Kanazawa UniversityLaboratory of Clinical Pharmacy, Kyoto Pharmaceutical UniversityDepartment of Biochemistry and Molecular Cell Biology, School of Medicine, College of Medicine, Taipei Medical UniversitySchool of Pharmacy, College of Medicine, National Taiwan UniversitySchool of Pharmacy, College of Medicine, National Taiwan UniversitySchool of Pharmacy, College of Medicine, National Taiwan UniversityAbstract Doxorubicin (DOX) is a potent chemotherapeutic widely used against various cancers, but its clinical application is limited by DOX-induced cardiotoxicity (DIC). This study explored the cardioprotective potential of extracellular vesicle-enriched secretome derived from adipose stem cells (EVSASC) in mitigating DOX-induced apoptosis in cardiomyocytes. Adipose-derived stem cells were cultured, and their conditioned medium and extraceullular vesicles were isolated and characterized according to the Minimal Information for Studies of Extracellular Vesicles 2023 guidelines. HL-1 cardiomyocytes were pretreated with EVSASC before exposure to 1 µM DOX. Cell viability was assessed via the cell counting kit-8 assay, while apoptosis markers and survival mediators were evaluated through Western blotting. RNA sequencing identified differentially expressed genes, including clusterin (Clu), which was further quantified using an enzyme-linked immunosorbent assay. The functional role of clusterin was validated through siRNA-mediated knockdown. EVSASC significantly improved cell viability in DOX-exposed cardiomyocytes and reduced the cleaved caspase-3 to procaspase-3 ratio. Clusterin expression was highest in EVSASC-treated cells, and its knockdown markedly increased caspase-3 cleavage, confirming its pivotal role in cardioprotection. Moreover, EVSASC enhanced the phosphorylation of AKT, Bcl2-associated agonist of cell death, and glycogen synthase kinase-3β, implicating PI3K/AKT pathway activation in clusterin upregulation and anti-apoptotic effects. These findings demonstrate that EVSASC mitigates DOX-induced apoptosis in cardiomyocytes through clusterin upregulation and PI3K/AKT pathway activation. Clusterin is identified as a potential biomarker for evaluating EVSASC efficacy. While EVSASC shows promise as a cardioprotective strategy against DIC, further studies are needed to optimize its therapeutic safety by addressing potential oncogenic risks.https://doi.org/10.1186/s13062-025-00664-5
spellingShingle Wan-Tseng Hsu
Shinji Kobuchi
Tung-Chun Russell Chien
I-Chun Chen
Shohei Hamada
Masayuki Tsujimoto
I-Lin Tsai
Yun-Sheng Wong
Kuan-Hsuan Tung
Ying-Zhen He
Extracellular vesicle-enriched secretome of adipose-derived stem cells upregulates clusterin to alleviate doxorubicin-induced apoptosis in cardiomyocytes
Biology Direct
title Extracellular vesicle-enriched secretome of adipose-derived stem cells upregulates clusterin to alleviate doxorubicin-induced apoptosis in cardiomyocytes
title_full Extracellular vesicle-enriched secretome of adipose-derived stem cells upregulates clusterin to alleviate doxorubicin-induced apoptosis in cardiomyocytes
title_fullStr Extracellular vesicle-enriched secretome of adipose-derived stem cells upregulates clusterin to alleviate doxorubicin-induced apoptosis in cardiomyocytes
title_full_unstemmed Extracellular vesicle-enriched secretome of adipose-derived stem cells upregulates clusterin to alleviate doxorubicin-induced apoptosis in cardiomyocytes
title_short Extracellular vesicle-enriched secretome of adipose-derived stem cells upregulates clusterin to alleviate doxorubicin-induced apoptosis in cardiomyocytes
title_sort extracellular vesicle enriched secretome of adipose derived stem cells upregulates clusterin to alleviate doxorubicin induced apoptosis in cardiomyocytes
url https://doi.org/10.1186/s13062-025-00664-5
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