Human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferation

It remains a significant challenge to reactivate the cell cycle activity of adult mammalian cardiomyocytes (CMs). This study created a hypo-immunogenic human induced pluripotent stem cell (hiPSC) line using clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 gene editing to knockout β2...

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Main Authors: Yuhua Wei, Xiaoxiao Geng, Qing You, Yu Zhang, Fangfang Cao, Gunaseelan Narayanan, Thanh Nguyen, Xiaoyuan Chen, Jianyi Zhang, Lei Ye
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2025-08-01
Series:Bioactive Materials
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Online Access:http://www.sciencedirect.com/science/article/pii/S2452199X25001562
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author Yuhua Wei
Xiaoxiao Geng
Qing You
Yu Zhang
Fangfang Cao
Gunaseelan Narayanan
Thanh Nguyen
Xiaoyuan Chen
Jianyi Zhang
Lei Ye
author_facet Yuhua Wei
Xiaoxiao Geng
Qing You
Yu Zhang
Fangfang Cao
Gunaseelan Narayanan
Thanh Nguyen
Xiaoyuan Chen
Jianyi Zhang
Lei Ye
author_sort Yuhua Wei
collection DOAJ
description It remains a significant challenge to reactivate the cell cycle activity of adult mammalian cardiomyocytes (CMs). This study created a hypo-immunogenic human induced pluripotent stem cell (hiPSC) line using clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 gene editing to knockout β2-microglobulin in hiPSCs (B2MKOhiPSCs) for manufacturing nanovesicles (B2MKOhiPSC-NVs). Approximately 9500 B2MKOhiPSC-NVs were produced from a single B2MKOhiPSC. Proteomic analyses indicated that, compared to B2MKOhiPSCs, the cargos of B2MKOhiPSC-NVs were enriched in spindle and chromosomal proteins, as well as proteins that regulate the cell cycle and scavenge reactive oxygen species (ROS). When administrated to hiPSCs derived CMs (hiPSC-CMs), B2MKOhiPSC-NVs reduced lactate dehydrogenase leakage and apoptosis in hypoxia-cultured hiPSC-CMs through activating the AKT pathway, protected hiPSC-CMs from H2O2-induced damage by ROS scavengers in the NV cargo, increased hiPSC-CM proliferation via the YAP pathway, and were hypoimmunogenic when co-cultured with human CD8+ T cells or delivered to mice. Furthermore, when B2MKOhiPSC-NVs or 0.9 % NaCl were intramyocardially injected into mice after cardiac ischemia/reperfusion injury, cardiac function and infarct size, assessed 4 weeks later, were significantly improved in the B2MKOhiPSC-NV group, with increased mouse CM survival and cell cycle activity. Thus, the proteins in the B2MKOhiPSC-NV cargos convergently activated the AKT pathway, scavenged ROS to protect CMs, and upregulated YAP signaling to induce CM cell cycle activity. Thus, B2MKOhiPSC-NVs hold great potential for cardiac protection and regeneration.
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spelling doaj-art-744d7715852843f3bf0607b7e5166d3d2025-08-20T03:20:15ZengKeAi Communications Co., Ltd.Bioactive Materials2452-199X2025-08-015058560210.1016/j.bioactmat.2025.04.017Human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferationYuhua Wei0Xiaoxiao Geng1Qing You2Yu Zhang3Fangfang Cao4Gunaseelan Narayanan5Thanh Nguyen6Xiaoyuan Chen7Jianyi Zhang8Lei Ye9Department of Biomedical Engineering, School of Medicine and School of Engineering, University of Alabama at Birmingham, Birmingham, AL, 35294, USADepartment of Biomedical Engineering, School of Medicine and School of Engineering, University of Alabama at Birmingham, Birmingham, AL, 35294, USADepartment of Diagnostic Radiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 119074, SingaporeDepartment of Diagnostic Radiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 119074, SingaporeDepartment of Diagnostic Radiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 119074, SingaporeDuke-NUS Medical School, National University of Singapore, Singapore, 169857, SingaporeDepartment of Biomedical Engineering, School of Medicine and School of Engineering, University of Alabama at Birmingham, Birmingham, AL, 35294, USADepartment of Diagnostic Radiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 119074, Singapore; Department of Chemical and Biomolecular Engineering, College of Design and Engineering, National University of Singapore, Singapore, 117575, Singapore; Department of Biomedical Engineering, College of Design and Engineering, National University of Singapore, Singapore, 117575, Singapore; Department of Pharmacy and Pharmaceutical Sciences, Faculty of Science, National University of Singapore, Singapore, 117544, Singapore; Clinical Imaging Research Centre, Centre for Translational Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117599, Singapore; Nanomedicine Translational Research Programme, Centre for NanoMedicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117597, Singapore; Theranostics Center of Excellence (TCE), Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 138667, Singapore; Institute of Molecular and Cell Biology, Agency for Science, Technology, and Research (A*STAR), 61 Biopolis Drive, Proteos, Singapore, 138673, SingaporeDepartment of Biomedical Engineering, School of Medicine and School of Engineering, University of Alabama at Birmingham, Birmingham, AL, 35294, USA; Department of Medicine, Division of Cardiovascular Disease, School of Medicine, University of Alabama at Birmingham, Birmingham, AL, 35294, USA; Corresponding author. Department of Biomedical Engineering, The University of Alabama at Birmingham, Volker Hall, 1670 University Boulevard, Birmingham, 35233, AL, USA.Department of Biomedical Engineering, School of Medicine and School of Engineering, University of Alabama at Birmingham, Birmingham, AL, 35294, USA; Corresponding author. Department of Biomedical Engineering, University of Alabama at Birmingham, Volker Hall, 1670 University Boulevard, Birmingham, 35233, AL, USA.It remains a significant challenge to reactivate the cell cycle activity of adult mammalian cardiomyocytes (CMs). This study created a hypo-immunogenic human induced pluripotent stem cell (hiPSC) line using clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 gene editing to knockout β2-microglobulin in hiPSCs (B2MKOhiPSCs) for manufacturing nanovesicles (B2MKOhiPSC-NVs). Approximately 9500 B2MKOhiPSC-NVs were produced from a single B2MKOhiPSC. Proteomic analyses indicated that, compared to B2MKOhiPSCs, the cargos of B2MKOhiPSC-NVs were enriched in spindle and chromosomal proteins, as well as proteins that regulate the cell cycle and scavenge reactive oxygen species (ROS). When administrated to hiPSCs derived CMs (hiPSC-CMs), B2MKOhiPSC-NVs reduced lactate dehydrogenase leakage and apoptosis in hypoxia-cultured hiPSC-CMs through activating the AKT pathway, protected hiPSC-CMs from H2O2-induced damage by ROS scavengers in the NV cargo, increased hiPSC-CM proliferation via the YAP pathway, and were hypoimmunogenic when co-cultured with human CD8+ T cells or delivered to mice. Furthermore, when B2MKOhiPSC-NVs or 0.9 % NaCl were intramyocardially injected into mice after cardiac ischemia/reperfusion injury, cardiac function and infarct size, assessed 4 weeks later, were significantly improved in the B2MKOhiPSC-NV group, with increased mouse CM survival and cell cycle activity. Thus, the proteins in the B2MKOhiPSC-NV cargos convergently activated the AKT pathway, scavenged ROS to protect CMs, and upregulated YAP signaling to induce CM cell cycle activity. Thus, B2MKOhiPSC-NVs hold great potential for cardiac protection and regeneration.http://www.sciencedirect.com/science/article/pii/S2452199X25001562Pluripotent stem cellsNanovesiclesCell cycleMyocardial protectionRegeneration
spellingShingle Yuhua Wei
Xiaoxiao Geng
Qing You
Yu Zhang
Fangfang Cao
Gunaseelan Narayanan
Thanh Nguyen
Xiaoyuan Chen
Jianyi Zhang
Lei Ye
Human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferation
Bioactive Materials
Pluripotent stem cells
Nanovesicles
Cell cycle
Myocardial protection
Regeneration
title Human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferation
title_full Human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferation
title_fullStr Human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferation
title_full_unstemmed Human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferation
title_short Human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferation
title_sort human induced pluripotent stem cell derived nanovesicles for cardiomyocyte protection and proliferation
topic Pluripotent stem cells
Nanovesicles
Cell cycle
Myocardial protection
Regeneration
url http://www.sciencedirect.com/science/article/pii/S2452199X25001562
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