2. Academic Validation
  3. Efficient Scaling up EV-AAVs Production via Cellular Nanoporation for Familial Hypercholesterolaemia Therapy

Efficient Scaling up EV-AAVs Production via Cellular Nanoporation for Familial Hypercholesterolaemia Therapy

  • J Extracell Vesicles. 2025 Nov;14(11):e70186. doi: 10.1002/jev2.70186.
Yuting Yan 1 2 3,  Yi You 2 4 5,  Shuhong Ma 1,  Hui Yi 6,  Guangduo Chen 3,  Jie Ni 2,  Changyan Chen 2,  Wenyu Ke 2,  Lingying Li 2,  Rui Bai 2,  Yuqing Ran 2 7,  Wenjing Lu 8,  Min Zhu 1,  Yongshuai Zhang 1,  Jing Dai 1,  Man Qi 1,  Feng Lan 1 2,  Andrew S Lee 5,  Ran Zhang 9,  Xujie Liu 2,  Zhaoyang Chen 3
Affiliations

Affiliations

  • 1 Key Laboratory of Pluripotent Stem Cells in Cardiac Repair and Regeneration, State Key Laboratory of Cardiovascular Disease, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Fuwai Hospital, Beijing, China.
  • 2 State Key Laboratory of Cardiovascular Disease, Chinese Academy of Medical Sciences, Fuwai Hospital, Shenzhen, China.
  • 3 Department of Cardiology, Heart Center of Fujian Province, Fujian Medical University Union Hospital, Fujian Medical University Heart Center, Fuzhou, Fujian, China.
  • 4 Department of Cardiovascular Surgery of the First Affiliated Hospital & Institute for Cardiovascular Science, Suzhou Medical College, Soochow University, Suzhou, Jiangsu, China.
  • 5 School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen, China.
  • 6 Division of Pediatric Cardiology, Department of Pediatric Medicine, The Seventh Medical Center of Chinese PLA General Hospital, Beijing, China.
  • 7 Wenzhou Medical University Graduate School, Wenzhou, China.
  • 8 Beijing Laboratory for Cardiovascular Precision Medicine, The Key Laboratory of Biomedical Engineering for Cardiovascular Disease Research, Ministry of Education, Beijing Anzhen Hospital, Capital Medical University, Beijing, China.
  • 9 Department of Cardiovascular Medicine, Chinese PLA General Hospital & Chinese PLA Medical School, Beijing, China.
PMID: 41216903 DOI: 10.1002/jev2.70186
Abstract

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.

Keywords

LDLR; cellular nanoporation technology; extracellular vesicles‐encapsulated AAV; familial hypercholesterolaemia.

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