Tmprss2 specific miRNAs as promising regulators for SARS-CoV-2 entry checkpoint

Area of Science:

• Molecular Biology
• Virology
• Computational Biology

Background:

• TMPRSS2 (transmembrane serine protease 2) is crucial for SARS-CoV-2 cellular entry and viral pathogenesis.
• Targeting TMPRSS2 is a promising strategy for COVID-19 intervention.
• MicroRNAs (miRNAs) are regulators of gene expression with potential therapeutic applications.

Purpose of the Study:

• To computationally screen for miRNAs that target and suppress TMPRSS2.
• To validate the efficacy of selected miRNAs in reducing TMPRSS2 expression in vitro.
• To explore the potential of miRNA-mediated TMPRSS2 suppression for COVID-19 prevention.

Main Methods:

• Computational screening of 163 miRNAs against TMPRSS2 using three prediction algorithms.
• Analysis of miRNA-TMPRSS2 binding affinity using computational negative energy calculations.
• Selection of top-scoring miRNAs (hsa-miR-214, hsa-miR-98, hsa-miR-32) using the Sfold tool.
• In vitro transfection of selected miRNAs into Caco-2 cells to assess TMPRSS2 gene silencing.

Main Results:

• Eleven common miRNAs predicted to bind TMPRSS2 were identified.
• Computational analysis confirmed miRNA-TMPRSS2 interactions.
• Three miRNAs (hsa-miR-214, hsa-miR-98, hsa-miR-32) were selected based on high probability scores and accessibility.
• Transfection experiments confirmed TMPRSS2 gene silencing, with hsa-miR-32 showing maximum suppression.
• Exalted binding and suppression of TMPRSS2 by miRNAs were observed.

Conclusions:

• Specific miRNAs, particularly hsa-miR-32, effectively suppress TMPRSS2 expression.
• These miRNAs demonstrate potential as molecular tools for preventing SARS-CoV-2 transmission and replication.
• miRNA-mediated TMPRSS2 inhibition offers a novel strategy for early COVID-19 prevention.