Efficient Scaling up EV‐AAVs Production via Cellular Nanoporation for Familial Hypercholesterolaemia Therapy
Adeno-associated virus (AAV)-mediated gene therapies face critical clinical limitations, including immune-mediated neutralization by pre-existing antibodies and dose-dependent hepatotoxicity. Extracellular vesicle-encapsulated AAVs (EV-AAVs) offer a promising solution by shielding AAVs from antibody recognition, yet existing production methods remain inefficient and impractical for clinical application. Here, we developed a cellular nanoporation (CNP) platform that enables scalable, high-yield generation of EV-AAVs, achieving an approximately 11-fold increase in production efficiency compared with conventional methods. In LDLR-deficient murine models with pre-existing neutralizing antibodies (1:200), EV-AAV-LDLR at half the standard AAV dose robustly restored hepatic LDL receptor expression and attenuated atherosclerosis progression. Notably, EV-AAV exhibited superior immune evasion capabilities, maintaining 2.3-fold higher hepatic transduction efficiency than conventional AAV upon secondary dosing due to antibody shielding. Importantly, EV-AAV therapy markedly reduced hepatotoxicity, with serum AST/ALT levels comparable to saline-treated controls, thereby overcoming a critical safety barrier of high-dose AAV treatment. These results demonstrate CNP as a clinically translatable platform for scalable EV-AAV manufacturing, enabling effective multi-dose regimens while overcoming key immunological and toxicity barriers in liver-directed gene therapy for familial hypercholesterolaemia.
Authors
- Yi You
- Man Qi (ORCID: https://orcid.org/0009-0002-5818-8766)
- Feng Lan (ORCID: https://orcid.org/0000-0002-9038-4014)
- Guangduo Chen
- Shuhong Ma (ORCID: https://orcid.org/0000-0002-9271-8030)
- Andrew S. Lee (ORCID: https://orcid.org/0000-0001-9169-1481)
- Jie Ni (ORCID: https://orcid.org/0000-0002-8844-6132)
- Yuting Yan (ORCID: https://orcid.org/0000-0002-9355-2984)
- Xujie Liu (ORCID: https://orcid.org/0000-0001-8713-4287)
- Yongshuai Zhang (ORCID: https://orcid.org/0000-0001-8165-2446)
- Ran Zhang (ORCID: https://orcid.org/0000-0002-0921-078X)
- Zhaoyang Chen (ORCID: https://orcid.org/0000-0003-0554-7702)
- Rui Bai (ORCID: https://orcid.org/0000-0003-4982-7562)
- Min Zhu (ORCID: https://orcid.org/0000-0003-2282-8274)
- Wenjing Lu (ORCID: https://orcid.org/0000-0002-0444-8991)
- Hui Yi (ORCID: https://orcid.org/0000-0003-0501-0972)
- Jing Dai (ORCID: https://orcid.org/0009-0000-6866-0986)
- Wujian Ke (ORCID: https://orcid.org/0000-0003-4800-4302)
- Yuqing Ran
- Changyan Chen (ORCID: https://orcid.org/0000-0003-1546-4011)
- Lingying Li
Institutions
- Fujian Medical University (CN)
- Chinese Academy of Medical Sciences & Peking Union Medical College (CN)
- Peking University (CN)
- Wenzhou Medical University (CN)
- Chinese PLA General Hospital (CN)
- Soochow University (CN)
- Beijing Anzhen Hospital (CN)
- First Affiliated Hospital of Soochow University (CN)
- Fu Wai Hospital (CN)
- Union Hospital (CN)
Publication Details
- Journal
- Journal of Extracellular Vesicles
- Published
- 2025-11-01
- DOI
- https://doi.org/10.1002/jev2.70186
- Primary Topic
- Extracellular vesicles in disease
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Natural Science Foundation of China
- Chinese Academy of Sciences
- China Postdoctoral Science Foundation
- Chinese Academy of Medical Sciences
- General Hospital of People’s Liberation Army
- Fujian Medical University
- Shenzhen Fundamental Research Program
- Shenzhen Bay Laboratory
- National Key Research and Development Program of China