Neutralization of SARS-CoV-2 pseudovirus using ACE2-engineered extracellular vesicles

Canhao Wu; Qin Xu; Huiyuan Wang; Bin Tu; Jiaxin Zeng; Pengfei Zhao; Mingjie Shi; Hong Qiu; Yongzhuo Huang

doi:10.1016/j.apsb.2021.09.004

Neutralization of SARS-CoV-2 pseudovirus using ACE2-engineered extracellular vesicles

Acta Pharm Sin B. 2022 Mar;12(3):1523-1533. doi: 10.1016/j.apsb.2021.09.004. Epub 2021 Sep 9.

Authors

Canhao Wu^{1

2}, Qin Xu¹, Huiyuan Wang², Bin Tu^{2

3}, Jiaxin Zeng^{1

2}, Pengfei Zhao^{2

4}, Mingjie Shi², Hong Qiu², Yongzhuo Huang^{2

3

5

6}

Affiliations

¹ Artemisinin Research Center, First Clinical School, Guangzhou University of Chinese Medicine, Guangzhou 510450, China.
² State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
³ Zhongshan Institute for Drug Discovery, SIMM, CAS, Zhongshan 528437, China.
⁴ Center of Clinical Pharmacology, The Second Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou 310009, China.
⁵ NMPA Key Laboratory for Quality Research and Evaluation of Pharmaceutical Excipients, Shanghai 201203, China.
⁶ Taizhou University, School of Advanced Study, Institute of Natural Medicine and Health Product, Taizhou 318000, China.

Abstract

The spread of coronavirus disease 2019 (COVID-19) throughout the world has resulted in stressful healthcare burdens and global health crises. Developing an effective measure to protect people from infection is an urgent need. The blockage of interaction between angiotensin-converting enzyme 2 (ACE2) and S protein is considered an essential target for anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) drugs. A full-length ACE2 protein could be a potential drug to block early entry of SARS-CoV-2 into host cells. In this study, a therapeutic strategy was developed by using extracellular vesicles (EVs) with decoy receptor ACE2 for neutralization of SARS-CoV-2. The EVs embedded with engineered ACE2 (EVs-ACE2) were prepared; the EVs-ACE2 were derived from an engineered cell line with stable ACE2 expression. The potential effect of the EVs-ACE2 on anti-SARS-CoV-2 was demonstrated by both in vitro and in vivo neutralization experiments using the pseudovirus with the S protein (S-pseudovirus). EVs-ACE2 can inhibit the infection of S-pseudovirus in various cells, and importantly, the mice treated with intranasal administration of EVs-ACE2 can suppress the entry of S-pseudovirus into the mucosal epithelium. Therefore, the intranasal EVs-ACE2 could be a preventive medicine to protect from SARS-CoV-2 infection. This EVs-based strategy offers a potential route to COVID-19 drug development.

Keywords: ACE2; ACE2, angiotensin-converting enzyme 2; BSA, bovine albumin; COVID-19; EVs, extracellular vesicles; Extracellular vesicles; FBS, fetal bovine serum; Intranasal administration; NTA, nanoparticle tracking analysis; Neutralization; PAGE, polyacrylamide gel electrophoresis; Pseudovirus; RIPA, radio immunoprecipitation assay; RLU, relative luminescence units; S protein, spike protein; SARS-CoV-2; SDS, sodium dodecyl sulfate; Spike protein; TEM, transmission electron microscope; WB, western blot.