Enhanced B cell electroporation efficiency via inhibition of DNA-induced apoptosis and pyroptosis with pan-caspase inhibitor
B cells have immense potential as gene therapy carriers because of their ability to secrete therapeutic proteins and mediate immunological memory and tolerogenic functions. However, efficient gene delivery into primary B cells remains a challenge, as DNA electroporation frequently induces significan...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-09-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050125001378 |
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| author | Gui-Lin Yang Deng-Gao Wang Xiang Zhao De-Long Hao Zhu-Qin Zhang De-Pei Liu |
| author_facet | Gui-Lin Yang Deng-Gao Wang Xiang Zhao De-Long Hao Zhu-Qin Zhang De-Pei Liu |
| author_sort | Gui-Lin Yang |
| collection | DOAJ |
| description | B cells have immense potential as gene therapy carriers because of their ability to secrete therapeutic proteins and mediate immunological memory and tolerogenic functions. However, efficient gene delivery into primary B cells remains a challenge, as DNA electroporation frequently induces significant cell death. Here, we demonstrate that primary murine B cells undergo apoptosis and pyroptosis after DNA electroporation, which is driven primarily by activation of the cyclic GMP-AMP synthase stimulator of interferon genes (cGAS-STING) pathway in response to cytoplasmic double-stranded DNA (dsDNA). To overcome this, we pretreated B cells with a caspase inhibitor, which increased cell viability and improved DNA delivery and knock-in efficiency. Notably, compared with the inhibition of either apoptosis or pyroptosis alone, treatment with the pan-caspase inhibitor Boc-D-FMK resulted in a more significant improvement in cell viability and DNA electroporation efficiency. This approach significantly increased the expression and secretion of human erythropoietin (hEPO) both in vitro and in vivo. These findings not only elucidate the mechanisms underlying DNA-electroporation-induced cell death in B cells but also establish a strategy to optimize B-cell-based gene therapies and therapeutic protein production. |
| format | Article |
| id | doaj-art-b9e1539bb2de49b783e2b57bcdb54a41 |
| institution | DOAJ |
| issn | 2329-0501 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj-art-b9e1539bb2de49b783e2b57bcdb54a412025-08-20T02:58:30ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012025-09-0133310154210.1016/j.omtm.2025.101542Enhanced B cell electroporation efficiency via inhibition of DNA-induced apoptosis and pyroptosis with pan-caspase inhibitorGui-Lin Yang0Deng-Gao Wang1Xiang Zhao2De-Long Hao3Zhu-Qin Zhang4De-Pei Liu5State Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, ChinaState Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, ChinaState Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, ChinaState Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, ChinaState Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China; Medical Epigenetics Research Center, Chinese Academy of Medical Sciences, Beijing 100005, China; Haihe Laboratory of Cell Ecosystem, Tianjin 300301, China; Corresponding author: Zhu-Qin Zhang, State Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.State Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China; Medical Epigenetics Research Center, Chinese Academy of Medical Sciences, Beijing 100005, China; Haihe Laboratory of Cell Ecosystem, Tianjin 300301, China; Corresponding author: De-Pei Liu, State Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.B cells have immense potential as gene therapy carriers because of their ability to secrete therapeutic proteins and mediate immunological memory and tolerogenic functions. However, efficient gene delivery into primary B cells remains a challenge, as DNA electroporation frequently induces significant cell death. Here, we demonstrate that primary murine B cells undergo apoptosis and pyroptosis after DNA electroporation, which is driven primarily by activation of the cyclic GMP-AMP synthase stimulator of interferon genes (cGAS-STING) pathway in response to cytoplasmic double-stranded DNA (dsDNA). To overcome this, we pretreated B cells with a caspase inhibitor, which increased cell viability and improved DNA delivery and knock-in efficiency. Notably, compared with the inhibition of either apoptosis or pyroptosis alone, treatment with the pan-caspase inhibitor Boc-D-FMK resulted in a more significant improvement in cell viability and DNA electroporation efficiency. This approach significantly increased the expression and secretion of human erythropoietin (hEPO) both in vitro and in vivo. These findings not only elucidate the mechanisms underlying DNA-electroporation-induced cell death in B cells but also establish a strategy to optimize B-cell-based gene therapies and therapeutic protein production.http://www.sciencedirect.com/science/article/pii/S2329050125001378B cellelectroporationcell deathapoptosispyroptosiscGAS-STING |
| spellingShingle | Gui-Lin Yang Deng-Gao Wang Xiang Zhao De-Long Hao Zhu-Qin Zhang De-Pei Liu Enhanced B cell electroporation efficiency via inhibition of DNA-induced apoptosis and pyroptosis with pan-caspase inhibitor Molecular Therapy: Methods & Clinical Development B cell electroporation cell death apoptosis pyroptosis cGAS-STING |
| title | Enhanced B cell electroporation efficiency via inhibition of DNA-induced apoptosis and pyroptosis with pan-caspase inhibitor |
| title_full | Enhanced B cell electroporation efficiency via inhibition of DNA-induced apoptosis and pyroptosis with pan-caspase inhibitor |
| title_fullStr | Enhanced B cell electroporation efficiency via inhibition of DNA-induced apoptosis and pyroptosis with pan-caspase inhibitor |
| title_full_unstemmed | Enhanced B cell electroporation efficiency via inhibition of DNA-induced apoptosis and pyroptosis with pan-caspase inhibitor |
| title_short | Enhanced B cell electroporation efficiency via inhibition of DNA-induced apoptosis and pyroptosis with pan-caspase inhibitor |
| title_sort | enhanced b cell electroporation efficiency via inhibition of dna induced apoptosis and pyroptosis with pan caspase inhibitor |
| topic | B cell electroporation cell death apoptosis pyroptosis cGAS-STING |
| url | http://www.sciencedirect.com/science/article/pii/S2329050125001378 |
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