Dihydroartemisinin decreases pre-existing neutralizing antibodies against adeno-associated virus in challenged mice
IntroductionThe high prevalence of pre-existing neutralizing antibodies (NAbs) against adeno-associated virus (AAV) poses a major obstacle to in vivo gene therapy. Current immunosuppressive (IS) strategies, such as corticosteroids, are limited by toxicity and adverse effects. To explore safer altern...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2025-08-01
|
| Series: | Frontiers in Pharmacology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2025.1587135/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850044535023861760 |
|---|---|
| author | Jingjing Fang Enze Cui Jinyan Xie Xuxia Gao Yun He Ming Yang Sana Shaheen Zhengjun Zhou Shaolai Zhou Binbin Cheng Binbin Cheng Changquan Ling Changquan Ling Chen Ling Chen Ling Chen Ling |
| author_facet | Jingjing Fang Enze Cui Jinyan Xie Xuxia Gao Yun He Ming Yang Sana Shaheen Zhengjun Zhou Shaolai Zhou Binbin Cheng Binbin Cheng Changquan Ling Changquan Ling Chen Ling Chen Ling Chen Ling |
| author_sort | Jingjing Fang |
| collection | DOAJ |
| description | IntroductionThe high prevalence of pre-existing neutralizing antibodies (NAbs) against adeno-associated virus (AAV) poses a major obstacle to in vivo gene therapy. Current immunosuppressive (IS) strategies, such as corticosteroids, are limited by toxicity and adverse effects. To explore safer alternatives, we evaluated dihydroartemisinin (DHA), a synthetic derivative of artemisinin inspired by traditional Chinese medicine (TCM), as a potential IS agent.MethodsIn vivo experiments were conducted by administering DHA at either 30 or 210 days post-injection (PI) of rAAVDJ vectors. Anti-AAV NAb levels, transgene expression, and vector genome biodistribution were assessed. Flow cytometry was used to quantify CD20+ B cells, germinal center B cells, and plasma cells in the spleen. Splenic gene expression profiling, liver histology, and serum biochemical analyses were performed to evaluate immunological and safety responses. In vitro, the impact of DHA and its serum metabolites on rAAV infection efficiency was tested in HEK293 cells.ResultsDHA administration significantly reduced anti-AAV NAb levels without compromising transgene expression or vector genome distribution. DHA treatment resulted in a reduction of splenic CD20+ B cells, germinal center B cells, and plasma cells, alongside changes in splenic gene expression profiles. Liver histology and serum markers confirmed that DHA at 125 mg/kg/day did not induce hepatotoxicity. In vitro assays demonstrated that DHA and its blood metabolites did not interfere with rAAV infection of HEK293 cells across multiple serotypes.DiscussionThese findings suggest that DHA is a safe and effective agent for modulating humoral immune responses to AAV vectors. Our results provide proof-of-concept evidence supporting the use of TCM-derived compounds to address immunological barriers in gene therapy. |
| format | Article |
| id | doaj-art-bf25b992288846d08906e235c3dcd636 |
| institution | DOAJ |
| issn | 1663-9812 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj-art-bf25b992288846d08906e235c3dcd6362025-08-20T02:54:54ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-08-011610.3389/fphar.2025.15871351587135Dihydroartemisinin decreases pre-existing neutralizing antibodies against adeno-associated virus in challenged miceJingjing Fang0Enze Cui1Jinyan Xie2Xuxia Gao3Yun He4Ming Yang5Sana Shaheen6Zhengjun Zhou7Shaolai Zhou8Binbin Cheng9Binbin Cheng10Changquan Ling11Changquan Ling12Chen Ling13Chen Ling14Chen Ling15Oncology Department of Traditional Chinese Medicine, Changhai Hospital, Naval Medical University, Shanghai, ChinaOncology Department of Traditional Chinese Medicine, Changhai Hospital, Naval Medical University, Shanghai, ChinaState Key Laboratory of Genetics and Development of Complex Phenotypes and Engineering Research Center of Gene Technology (Ministry of Education), School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, ChinaState Key Laboratory of Genetics and Development of Complex Phenotypes and Engineering Research Center of Gene Technology (Ministry of Education), School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, ChinaState Key Laboratory of Genetics and Development of Complex Phenotypes and Engineering Research Center of Gene Technology (Ministry of Education), School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, ChinaState Key Laboratory of Genetics and Development of Complex Phenotypes and Engineering Research Center of Gene Technology (Ministry of Education), School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, ChinaState Key Laboratory of Genetics and Development of Complex Phenotypes and Engineering Research Center of Gene Technology (Ministry of Education), School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, ChinaDepartment of Liver Surgery and Transplantation, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, ChinaDepartment of Liver Surgery and Transplantation, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, ChinaOncology Department of Traditional Chinese Medicine, Changhai Hospital, Naval Medical University, Shanghai, ChinaFaculty of Traditional Chinese Medicine, Naval Medical University, Shanghai, ChinaOncology Department of Traditional Chinese Medicine, Changhai Hospital, Naval Medical University, Shanghai, ChinaFaculty of Traditional Chinese Medicine, Naval Medical University, Shanghai, ChinaState Key Laboratory of Genetics and Development of Complex Phenotypes and Engineering Research Center of Gene Technology (Ministry of Education), School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, ChinaShanghai Key Laboratory of Gene Editing and Cell Therapy for Rare Diseases, Fudan University, Shanghai, ChinaInstitutes of Integrative Medicine, Fudan University, Shanghai, ChinaIntroductionThe high prevalence of pre-existing neutralizing antibodies (NAbs) against adeno-associated virus (AAV) poses a major obstacle to in vivo gene therapy. Current immunosuppressive (IS) strategies, such as corticosteroids, are limited by toxicity and adverse effects. To explore safer alternatives, we evaluated dihydroartemisinin (DHA), a synthetic derivative of artemisinin inspired by traditional Chinese medicine (TCM), as a potential IS agent.MethodsIn vivo experiments were conducted by administering DHA at either 30 or 210 days post-injection (PI) of rAAVDJ vectors. Anti-AAV NAb levels, transgene expression, and vector genome biodistribution were assessed. Flow cytometry was used to quantify CD20+ B cells, germinal center B cells, and plasma cells in the spleen. Splenic gene expression profiling, liver histology, and serum biochemical analyses were performed to evaluate immunological and safety responses. In vitro, the impact of DHA and its serum metabolites on rAAV infection efficiency was tested in HEK293 cells.ResultsDHA administration significantly reduced anti-AAV NAb levels without compromising transgene expression or vector genome distribution. DHA treatment resulted in a reduction of splenic CD20+ B cells, germinal center B cells, and plasma cells, alongside changes in splenic gene expression profiles. Liver histology and serum markers confirmed that DHA at 125 mg/kg/day did not induce hepatotoxicity. In vitro assays demonstrated that DHA and its blood metabolites did not interfere with rAAV infection of HEK293 cells across multiple serotypes.DiscussionThese findings suggest that DHA is a safe and effective agent for modulating humoral immune responses to AAV vectors. Our results provide proof-of-concept evidence supporting the use of TCM-derived compounds to address immunological barriers in gene therapy.https://www.frontiersin.org/articles/10.3389/fphar.2025.1587135/fullgene therapyadeno-associated virus vectorneutralizing antibodyimmunosuppression agentstraditional Chinese medicinedihydroartemisinin |
| spellingShingle | Jingjing Fang Enze Cui Jinyan Xie Xuxia Gao Yun He Ming Yang Sana Shaheen Zhengjun Zhou Shaolai Zhou Binbin Cheng Binbin Cheng Changquan Ling Changquan Ling Chen Ling Chen Ling Chen Ling Dihydroartemisinin decreases pre-existing neutralizing antibodies against adeno-associated virus in challenged mice Frontiers in Pharmacology gene therapy adeno-associated virus vector neutralizing antibody immunosuppression agents traditional Chinese medicine dihydroartemisinin |
| title | Dihydroartemisinin decreases pre-existing neutralizing antibodies against adeno-associated virus in challenged mice |
| title_full | Dihydroartemisinin decreases pre-existing neutralizing antibodies against adeno-associated virus in challenged mice |
| title_fullStr | Dihydroartemisinin decreases pre-existing neutralizing antibodies against adeno-associated virus in challenged mice |
| title_full_unstemmed | Dihydroartemisinin decreases pre-existing neutralizing antibodies against adeno-associated virus in challenged mice |
| title_short | Dihydroartemisinin decreases pre-existing neutralizing antibodies against adeno-associated virus in challenged mice |
| title_sort | dihydroartemisinin decreases pre existing neutralizing antibodies against adeno associated virus in challenged mice |
| topic | gene therapy adeno-associated virus vector neutralizing antibody immunosuppression agents traditional Chinese medicine dihydroartemisinin |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2025.1587135/full |
| work_keys_str_mv | AT jingjingfang dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT enzecui dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT jinyanxie dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT xuxiagao dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT yunhe dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT mingyang dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT sanashaheen dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT zhengjunzhou dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT shaolaizhou dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT binbincheng dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT binbincheng dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT changquanling dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT changquanling dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT chenling dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT chenling dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice AT chenling dihydroartemisinindecreasespreexistingneutralizingantibodiesagainstadenoassociatedvirusinchallengedmice |