miR-98 Regulates TMPRSS2 Expression in Human Endothelial Cells: Key Implications for COVID-19

Abstract

The two main co-factors needed by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to enter human cells are angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2). Here, we focused on the study of microRNAs that specifically target TMPRSS2. Through a bioinformatic approach, we identified miR-98-5p as a suitable candidate. Since we and others have shown that endothelial cells play a pivotal role in the pathogenesis of the coronavirus disease 2019 (COVID-19), we mechanistically validated miR-98-5p as a regulator of TMPRSS2 transcription in two different human endothelial cell types, derived from the lung and from the umbilical vein. Taken together, our findings indicate that TMPRSS2 represents a valid target in COVID-19 treatment, which may be achieved by specific non-coding-RNA approaches.

miR-24 Targets the Transmembrane Glycoprotein Neuropilin-1 in Human Brain Microvascular Endothelial Cells

“…To identify miRNAs targeting the 3′-UTR of Neuropilin-1, we used the online target prediction tool Targetscan 7.2, as previously described by our research group [ 28 , 30 , 31 , 32 , 33 ].…”

“…To evaluate the effects of miR-24 on Neuropilin-1 gene transcription, we used a luciferase reporter containing the 3’-UTR of the predicted miRNA interaction site, both wild-type and mutated, in hBMECs cells. The mutant construct of Neuropilin-1 3′-UTR (Neuropilin-1 MUT, as shown in Figure 1 ), harboring a substitution of two nucleotides within the predicted miR-24 binding sites of Neuropilin-1 3′-UTR was obtained through means of the NEBaseChanger and Q5 site-directed mutagenesis kit (New England Biolabs, Ipswich, MA, USA) as we described [ 30 , 32 ].…”

“…We transfected hBMECs with the 3′-UTR reporter plasmid (0.05 μg) and miR-24 mirVana TM mimics (ThermoFisher Scientific, Waltham MA, USA) or miR-24 miRIDIAN hairpin inhibitors (PerkinElmer, Waltham MA, USA), as well as a non-targeting negative control (scramble), all used at a final concentration of 50 nMol/L, using Lipofectamine RNAiMAX (ThermoFisher Scientific) [ 32 ]. Firefly and Renilla luciferase activities were measured 48 h after transfection, using a Luciferase Reporter Assay System (Promega, Madison, WI, USA), normalizing Firefly luciferase to Renilla luciferase activity [ 32 ].…”

Interactions between hypertension and inflammatory tone and the effect on blood pressure and outcomes in patients with COVID‐19

“…Hence, it has been shown that endothelial involvement is associated with microvascular thrombi and prothrombotic state in patients with COVID-19. [28][29][30] Also, in a series of infected patients with SARS-COV2, evidence has been reported of direct viral infection of the endothelial cell and diffuse endothelial inflammation. 31 The current study revealed that a high level of infection-induced inflammation at the time of admission predicts a poor prognosis in patients with hypertension.…”

MicroRNAs and SARS-CoV-2 life cycle, pathogenesis, and mutations: biomarkers or therapeutic agents?

“…In addition, host miRNAs might block attachment and entry of SARS-CoV-2 because several miRNA families were found to target ACE2 and TMPRSS2 genes in different tissues. In an in vitro study, hsa-miR-98-5p was demonstrated to directly target the 3' UTR of TMPRSS2 gene transcription, both in human lung microvascular endothelial cells (HMVEC-L) and human umbilical vein endothelial cells (HUVEC) [29]. The hsa-let-7e/hsa-mir-125a and hsa-mir-141/hsa-miR-200 miRNA families inhibit ACE2/TMPRSS2 gene transcription.…”