Reporter Virus-Based High-Throughput Screening Identifies LM-10 as a Novel Inhibitor Targeting KRM1 for Coxsackievirus A10 Blockade
5 affiliations
- 1Key Laboratory of Marine Drugs Ministry of Education, School of Medicine and Pharmacy, Sanya Oceanographic Institution, Ocean University of China, Qingdao 266003, China
- 2Capital Center for Children’s Health, Capital Medical University, Capital Institute of Pediatrics, Beijing 100020, China
- 3Qingdao Center of Technology Innovation for Shark Antibody Development, College of Biological Engineering, Qingdao University of Science and Technology, Qingdao 266042, China
- 4CAS Key Laboratory of Pathogen Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
- 5State Key Laboratory of Biopharmaceutical Preparation and Delivery, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
Abstract
Enteroviruses causing hand, foot, and mouth disease, including currently circulating Coxsackievirus A10 (CVA10) and Coxsackievirus A6 (CVA6), rely on KREMEN1 (KRM1) for cell entry, making the virus–KRM1 interface a potential target for antiviral intervention. We screened GLMarNP, an in-house library of about 3000 natural products and derivatives. Candidates targeting the KRM1 ectodomain were selected by blind docking, drug-likeness assessment, and scaffold diversity analysis. To enable high-throughput antiviral screening, we inserted eGFP into a full-length infectious CVA10 cDNA clone to generate a reporter virus designated eGFP-A10. The virus was rescued in HEK293T cells and produced robust fluorescence in RD cells over serial passages. Among 29 candidate compounds, LM-10 showed the strongest antiviral activity and exhibited low cytotoxicity. Mechanistically, LM-10 bound KRM1 and acted early in the viral life cycle. Molecular docking suggested that LM-10 binds the KR domain at a site overlapping the VP2 EF-loop contact region. KRM1 also acts as a DKK1 receptor to regulate Wnt/β-catenin signaling. LM-10 did not significantly affect this pathway in the TOP-GFP reporter assay or in transgenic zebrafish embryos. These findings establish a practical CVA10 reporter-virus platform and support steric blockade of the receptor interface as a potential antiviral strategy.
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BibTeX
@article{li2026reporter,
title = {{Reporter Virus-Based High-Throughput Screening Identifies LM-10 as a Novel Inhibitor Targeting KRM1 for Coxsackievirus A10 Blockade}},
author = {Haimeng Li and Wenrui Gai and Hailan Yao and Yuchao Gu and Yuyang Li and Minlong Chen and Guangyuan Luo and Jianfeng Zhou and Yu Tang and Zhengyang Wang and Dehai Li and Yingzi Cui},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27219460},
url = {https://www.mdpi.com/1422-0067/27/21/9460},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Reporter Virus-Based High-Throughput Screening Identifies LM-10 as a Novel Inhibitor Targeting KRM1 for Coxsackievirus A10 Blockade
AU - Haimeng Li
AU - Wenrui Gai
AU - Hailan Yao
AU - Yuchao Gu
AU - Yuyang Li
AU - Minlong Chen
AU - Guangyuan Luo
AU - Jianfeng Zhou
AU - Yu Tang
AU - Zhengyang Wang
AU - Dehai Li
AU - Yingzi Cui
PY - 2026
DA - 2026/10/23
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27219460
UR - https://www.mdpi.com/1422-0067/27/21/9460
AB - Enteroviruses causing hand, foot, and mouth disease, including currently circulating Coxsackievirus A10 (CVA10) and Coxsackievirus A6 (CVA6), rely on KREMEN1 (KRM1) for cell entry, making the virus–KRM1 interface a potential target for antiviral intervention. We screened GLMarNP, an in-house library of about 3000 natural products and derivatives. Candidates targeting the KRM1 ectodomain were selected by blind docking, drug-likeness assessment, and scaffold diversity analysis. To enable high-throughput antiviral screening, we inserted eGFP into a full-length infectious CVA10 cDNA clone to generate a reporter virus designated eGFP-A10. The virus was rescued in HEK293T cells and produced robust fluorescence in RD cells over serial passages. Among 29 candidate compounds, LM-10 showed the strongest antiviral activity and exhibited low cytotoxicity. Mechanistically, LM-10 bound KRM1 and acted early in the viral life cycle. Molecular docking suggested that LM-10 binds the KR domain at a site overlapping the VP2 EF-loop contact region. KRM1 also acts as a DKK1 receptor to regulate Wnt/β-catenin signaling. LM-10 did not significantly affect this pathway in the TOP-GFP reporter assay or in transgenic zebrafish embryos. These findings establish a practical CVA10 reporter-virus platform and support steric blockade of the receptor interface as a potential antiviral strategy.
SN - 1422-0067
ER -