Pharma Focus Asia

Developing Inhibitory Peptides Against SARS-CoV-2 Envelope Protein

Ramsey Bekdash, Kazushige Yoshida, Manoj S. Nair, Lauren Qiu, Johnathan Ahdout, Hsiang-Yi Tsai, Kunihiro Uryu, Rajesh K. Soni, Yaoxing Huang, David D. Ho, Masayuki Yazawa.

Abstract

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has affected approximately 800 million people since the start of the Coronavirus Disease 2019 (COVID-19) pandemic. Because of the high rate of mutagenesis in SARS-CoV-2, it is difficult to develop a sustainable approach for prevention and treatment. The Envelope (E) protein is highly conserved among human coronaviruses.

Introduction

The Coronavirus Disease 2019 (COVID-19) pandemic caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has affected approximately 800 million people and counting in the world. More than 6 million people have passed away due to the viral infection. Because the mutagenesis rate in SARS-CoV-2 genes such as Spike is high, it is a challenge to develop sustainable approaches for prevention and treatment. While several new vaccines and drug candidates have become available, the number of COVID-19 infections and deaths are still increasing, and new variants are being reported.

Methods:

Columbia University is a National Institute of Health (NIH) Office of Laboratory Animal Welfare assured institution (ID#, D16-00003/A3007-01), which has complied with the NIH Public Health Service Policy and adhered to the standards in the guide for the care and use of laboratory animals. The animal study is approved by Columbia University Institutional Animal Care and Use Committee (protocol #, AC-AABP2571). This virus study using SARS-CoV-2 is approved by Columbia University ADARC committee and BSL3 facility committee.

Discussion:

We applied a pH fluorescent dye, DND-189, to identify TAT-MY18 as a therapeutic candidate against E proteins. However, uptake of the dye could be dependent on cell viability and intracellular organelle function because DND-189 is a single green fluorescent dye. Therefore, the fluorescent change might not be due to a direct effect of E protein, but a secondary effect. In addition to the DND-189 dye, we applied the NFAT/AP-1 luciferase reporter assay as a higher throughput screening platform to optimize the MY18 peptide using molecular biological approach.

Acknowledgments:

We thank N. Harrison, R. Katz, M. Rahmany, C. Y.l. Sobolevsky, M.V. Yelshanskaya, C. Aston, E. Passague, and J. Stein (Columbia University) for their helpful support and discussion; Y. Tomono and K. Yamamoto (Shigei Medical Research Institute, Japan) for helpful advice with rat monoclonal antibody production; C. Castagna, Y. Luo, S. Sozomenu, A. Matveyenko and M.H. Blumenkrantz (Columbia University) and A. Poddar (Peddie High School, NJ) for helpful assistance.

Citation: Bekdash R, Yoshida K, Nair MS, Qiu L, Ahdout J, Tsai H-Y, et al. (2024) Developing inhibitory peptides against SARS-CoV-2 envelope protein. PLoS Biol 22(3): e3002522. https://doi.org/10.1371/journal.pbio.3002522

Editor: Frank Kirchhoff, Ulm University Medical Center, GERMANY

Received: September 18, 2023; Accepted: January 25, 2024; Published: March 14, 2024.

Copyright: © 2024 Bekdash et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data are within the paper and its Supporting Information files. The data can be also accessible through Icahn School of Medicine at Mount Sinai Data Management and Research Compliance Committee. All materials and plasmid constructs used in this study have been maintained by Dr. Yazawa’s laboratory and are available upon request. In the future, the constructs will be deposited and available to order through Addgene.

Funding: This work was supported by Columbia University Dean’s Office Fund and Columbia University Translational Therapeutics (TRx) Pilot Award (to M.Y.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: M.Y., R.B., K.Y., D.D.H., M.S.N., and Y.H. (inventors) filed a patent (Attorney Docket No.: 01001/00889-US0; status: Filed, 04/13/2022) related to this manuscript. This patent is for using synthetic peptides targeting SARS-CoV-2 envelope protein for treating COVID-19 and related human coronaviruses. The rest of the authors declare no competing interests.

Abbreviations: BSA, bovine serum albumin; COVID-19, Coronavirus Disease 2019; DMEM, Dulbecco’s Modified Eagle Media; DMSO, Dimethylsulfoxide; ECL, enhanced chemiluminescence; ER, endoplasmic reticulum; ERGIC, ER-Golgi inter-compartment; FBS, fetal bovine serum; FDR, false discovery rate; HEK, human embryonic kidney; KLH, keyhole limpet haemocyanin; MERS-CoV, Middle East Respiratory Syndrome Coronavirus; NCE, normalized collision energy; NIH, National Institute of Health; PBS, phosphate-buffered saline; PCR, polymerase chain reaction; PS, penicillin/streptomycin; PVDF, polyvinylidene difluoride; SARS‑CoV‑2, Severe Acute Respiratory Syndrome Coronavirus 2; SPS, synchronous precursor selection; TBS-T, Tris-buffered saline with 0.1% Tween 20; TMT, tandem mass tag.

 

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