Leveraging Artificial Intelligence and Gene Expression Analysis to Identify Some Potential Bovine Coronavirus (BCoV) Receptors and Host Cell Enzymes Potentially Involved in the Viral Replication and Tissue Tropism

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(3), P. 1328 - 1328

Published: Feb. 4, 2025

Bovine coronavirus (BCoV) exhibits dual tissue tropism, infecting both the respiratory and enteric tracts of cattle. Viral entry into host cells requires a coordinated interaction between viral proteins. However, specific cellular receptors co-receptors facilitating BCoV remain poorly understood. Similarly, roles proteases such as Furin, TMPRSS2, Cathepsin-L (CTS-L), known to assist in replication other coronaviruses, have not been extensively explored for BCoV. This study aims identify novel that modulate tropism. cell lines were infected with isolates from origins, gene expression profiles post-infection analyzed using next-generation sequencing (NGS). Differentially expressed genes encoding potential further assessed in-silico prediction molecular docking analysis. These analyses focused on receptors, including ACE2, NRP1, DPP4, APN, AXL, CEACAM1, their infection. Validation these findings was performed qRT-PCR assays targeting individual genes. We confirmed enzymes some (+/−) lung tissues. Results revealed high binding affinities 9-O-acetylated sialic acid NRP1 spike (S) hemagglutinin-esterase (HE) proteins compared CEACAM1. Additionally, Furin TMPRSS2 predicted interact BCoV-S polybasic cleavage site (RRSRR|A), suggesting S glycoprotein activation. is first explore interactions multiple proteases. Functional studies are recommended confirm infection replication.

Language: Английский

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The dual actions of host miRNA-16a in restricting bovine coronavirus (BCoV) replication through targeting the host cell Furin and enhancing the host immune response

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: June 14, 2024

The roles of host cell miRNAs have not been well studied in the context BCoV replication and immune regulation. main aim this study was to identify miRNA candidates that regulate essential genes involved replication, tissue tropism, To achieve these goals, we used two isolates (enteric respiratory) infect bovine endothelial cells (BECs) Madine Darby Bovine Kidney (MDBK) cells. This is addition ex vivo model using peripheral blood mononuclear (PBMCs). We determined expression profiles after infection. miR-16a differentially altered during Our data show miRNA-16a a significantly downregulated both vitro ex vivo models. confirmed profile by qRT‒PCR. Overexpression pre-miRNA-16a BEC MDBK lines markedly inhibited infection, as viral genome copy numbers measured qRT‒PCR, protein (S N) Western blot, virus infectivity plaque assay. bioinformatic prediction showed Furin potential target miRNA-16a. compared level pre-miRNA-16a-transfected/BCoV-infected pre-miRNA-scrambled-transfected qRT‒PCR blot revealed marked inhibition at mRNA level, respectively. BCoV-S levels. further confirm impact downregulation enzyme on BCoV, transfected with specific Furin-siRNAs parallel scrambled siRNA. Marked observed Furin-siRNA-treated group. validate novel for miRNA-16a, cloned 3'UTR carrying seed region dual luciferase vector. activity pre-miRNA-16a-transfected decreased more than 50% construct mutated region. inhibits targeting line glycoprotein. It also enhances response, which contributes replication. our knowledge, first valid findings highlight clinical applications miRNA-based vaccine/antiviral therapy.

Language: Английский

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Identification of Some Potential Cellular Receptors and Host Enzymes That Could Potentially Refine the Bovine Coronavirus (BCoV) Replication and Tissue Tropism. A Molecular Docking Study.

Published: July 2, 2024

Bovine coronavirus (BCoV) is one of the most common pathogens affecting cattle all ages. BCoV possesses multiple tissue tropism in cattle, particularly respiratory and enteric tracts affected animals. Viral entry a crucial step replication cycle viruses, including BCoV. The process viral requires orchestration between several proteins from virus host side. spike glycoproteins contributed substantially to other infections molecular pathogenesis coronaviruses, Meanwhile, among corona-viruses, hemagglutinin esterase enzyme found only human corona-virus-OC43 (HCoV-OC43). receptors are key players could explain tropism, at least part. presence some cell proteases also en-hances until release cells. Previous studies have shown that 9- O-acetylated sialic acids potentially act as for However, little known about if co/receptors might be needed ensure In addition that, roles proteases, such Furin TMPRSS2, infection not yet been well studied. main objectives current study identify novel enhance replication. We used silico docking tools predict new out (ACE-2, NRP1, DPP4, APN, CEACAM1). Our results postulated potential actions TMPRRS2 enzymes activation BCoV-S glycoprotein, which enhances modeling confirms high binding affinities both NRP1 with BCoV-HE compared ACE-2, CEACAM-1, suggesting their prediction models show bindings TMPRSS2 polybasic cleavage site (RRSRR|A). To our knowledge, this first investigate bovine interaction receptors, APN CEACAM1, protease.

Language: Английский

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The dual actions of miRNA16a in restricting Bovine Coronavirus replication through downregulation of Furin and enhancing the host immune response

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: Nov. 26, 2024

The roles of host cell miRNAs have not been well studied in the context BCoV replication and immune regulation. This study aimed to identify miRNA candidates that regulate essential genes involved replication, tissue tropism, To achieve these goals, we used two isolates (enteric respiratory) infect bovine endothelial cells (BECs) Madine Darby Bovine Kidney (MDBK) cells. We determined expression profiles after infection. miRNA16a is differentially altered during Our data show a significantly downregulated both vitro ex vivo models. confirmed miRNA16aexpression profile by qRT–PCR. Overexpression pre-miRNA16ain BEC MDBK lines markedly inhibited infection, as viral genome copy numbers measured qRT‒PCR, protein (S N) Western blot, virus infectivity using plaque assay. bioinformatic prediction showed Furin potential target miRNA16a. compared level pre-miRNA16a-transfected/BCoV-infected pre-miRNA-scrambled-transfected qRT-PCR blot revealed marked inhibition at mRNA levels, respectively. BCoV-S was levels. further confirm impact downregulation enzyme on BCoV, transfected with specific Furin-siRNAs parallel scrambled siRNA. Marked observed Furin-siRNA-treated group. validate novel for miRNA16a, cloned 3'UTR carrying seed region dual luciferase vector. activity pre-miRNA16a-transfected decreased more than 50% construct mutated region. miRNA16ainhibits targeting line glycoprotein. It also enhances response, which contributes replication. first valid findings highlight clinical applications miRNA-based vaccine/antiviral therapy.

Language: Английский

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The dual actions of the host miRNA-16a in restricting Bovine coronavirus (BCoV) replication through targeting host cell Furin and in enhancing the host immune response.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 29, 2024

1. Abstract The roles of host cell miRNAs have not been studied well in the context BCoV replication and immune regulation. main aim this study was to identify some miRNA candidates that regulate essential genes involved replication, tissue tropism, To achieve these goals, we used two isolates (enteric respiratory) infect bovine endothelial cells (BEC) Madine Darby Bovine Kidney (MDBK) cells. This is addition ex vivo model using peripheral blood mononuclear (PBMC). We determined expression profiles after infection. miRA-16a one differentially altered during Our data shows miRNA-16a a significantly downregulated both vitro models. confirmed profile by qRT-PCR. Overexpression pre-miRNA-16a BEC MDBK lines resulted marked inhibition infection based on viral genome copy numbers measured qRT-PCR, protein (S N) Western blot, virus infectivity plaque assay. bioinformatic prediction showed Furin potential target for miRNA-16a. checked level transfected/BCoV infected compared pre-miRNA scrambled validate that. levels mRNA qRT-PCR blot. BCoV-S markedly inhibited levels. further confirm impacts downregulation enzyme BCoV, transfected with specific Furin-siRNA parallel siRNA. A observed Furin-siRNA-treated group. as novel miRNA-16a, cloned 3’UTR carrying seed region dual luciferase vector. activity decreased more than 50% construct mutated region. confirms inhibits targeting glycoprotein. It will also enhance response, which contributes replication. our knowledge, first valid findings highlight clinical applications miRNA-based vaccine/antiviral therapy.

Language: Английский

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