학술논문
Strategies to target SARS-CoV-2 entry and infection using dual mechanisms of inhibition by acidification inhibitors.
Document Type
Academic Journal
Author
Prabhakara C; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Godbole R; National Centre for Biological Sciences (TIFR), Bengaluru, India.; University of Trans-Disciplinary Health Sciences and Technology (TDU), Bengaluru, India.; Sil P; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Jahnavi S; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Gulzar SE; National Centre for Biological Sciences (TIFR), Bengaluru, India.; SASTRA University, Thanjavur, India.; van Zanten TS; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Sheth D; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Subhash N; National Centre for Biological Sciences (TIFR), Bengaluru, India.; SASTRA University, Thanjavur, India.; Chandra A; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Shivaraj A; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Panikulam P; National Centre for Biological Sciences (TIFR), Bengaluru, India.; U I; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Nuthakki VK; CSIR-Indian Institute of Integrative Medicine, Jammu, India.; Puthiyapurayil TP; Institute for Stem Cell Science and Regenerative Medicine (inSTEM), Bengaluru, India.; Ahmed R; CSIR-Indian Institute of Integrative Medicine, Jammu, India.; Najar AH; CSIR-Indian Institute of Integrative Medicine, Jammu, India.; Lingamallu SM; Institute for Stem Cell Science and Regenerative Medicine (inSTEM), Bengaluru, India.; Manipal Academy of Higher Education (MAHE), Madhav Nagar, Manipal, Karnataka, India.; Das S; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Mahajan B; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Vemula P; Institute for Stem Cell Science and Regenerative Medicine (inSTEM), Bengaluru, India.; Bharate SB; CSIR-Indian Institute of Integrative Medicine, Jammu, India.; Singh PP; CSIR-Indian Institute of Integrative Medicine, Jammu, India.; Vishwakarma R; CSIR-Indian Institute of Integrative Medicine, Jammu, India.; Guha A; Institute for Stem Cell Science and Regenerative Medicine (inSTEM), Bengaluru, India.; Sundaramurthy V; National Centre for Biological Sciences (TIFR), Bengaluru, India.; Mayor S; National Centre for Biological Sciences (TIFR), Bengaluru, India.
Source
Publisher: Public Library of Science Country of Publication: United States NLM ID: 101238921 Publication Model: eCollection Cited Medium: Internet ISSN: 1553-7374 (Electronic) Linking ISSN: 15537366 NLM ISO Abbreviation: PLoS Pathog Subsets: MEDLINE
Subject
Language
English
Abstract
Many viruses utilize the host endo-lysosomal network for infection. Tracing the endocytic itinerary of SARS-CoV-2 can provide insights into viral trafficking and aid in designing new therapeutic strategies. Here, we demonstrate that the receptor binding domain (RBD) of SARS-CoV-2 spike protein is internalized via the pH-dependent CLIC/GEEC (CG) endocytic pathway in human gastric-adenocarcinoma (AGS) cells expressing undetectable levels of ACE2. Ectopic expression of ACE2 (AGS-ACE2) results in RBD traffic via both CG and clathrin-mediated endocytosis. Endosomal acidification inhibitors like BafilomycinA1 and NH4Cl, which inhibit the CG pathway, reduce the uptake of RBD and impede Spike-pseudoviral infection in both AGS and AGS-ACE2 cells. The inhibition by BafilomycinA1 was found to be distinct from Chloroquine which neither affects RBD uptake nor alters endosomal pH, yet attenuates Spike-pseudovirus entry. By screening a subset of FDA-approved inhibitors for functionality similar to BafilomycinA1, we identified Niclosamide as a SARS-CoV-2 entry inhibitor. Further validation using a clinical isolate of SARS-CoV-2 in AGS-ACE2 and Vero cells confirmed its antiviral effect. We propose that Niclosamide, and other drugs which neutralize endosomal pH as well as inhibit the endocytic uptake, could provide broader applicability in subverting infection of viruses entering host cells via a pH-dependent endocytic pathway.
Competing Interests: The authors have declared that no competing interests exist.
Competing Interests: The authors have declared that no competing interests exist.