Effect of Ivermectin and Atorvastatin on Nuclear Localization of Importin Alpha and Drug Target Expression Profiling in Host Cells from Nasopharyngeal Swabs of SARS-CoV-2- Positive Patients.

Valeria Inés Segatori,Pablo Sanchis,Alejandro Krolewiecki,Rosario Lavignolle,Lorena Grisel Caligiuri,Juan Bizzotto,Juan Garona,Eduardo Spitzer,Ayelén Toro,Geraldine Gueron,Daniel Fernando Alonso

doi:10.3390/v13102084

Abstract

Nuclear transport and vesicle trafficking are key cellular functions involved in the pathogenesis of RNA viruses. Among other pleiotropic effects on virus-infected host cells, ivermectin (IVM) inhibits nuclear transport mechanisms mediated by importins and atorvastatin (ATV) affects actin cytoskeleton-dependent trafficking controlled by Rho GTPases signaling. In this work, we first analyzed the response to infection in nasopharyngeal swabs from SARS-CoV-2-positive and -negative patients by assessing the gene expression of the respective host cell drug targets importins and Rho GTPases. COVID-19 patients showed alterations in KPNA3, KPNA5, KPNA7, KPNB1, RHOA, and CDC42 expression compared with non-COVID-19 patients. An in vitro model of infection with Poly(I:C), a synthetic analog of viral double-stranded RNA, triggered NF-κB activation, an effect that was halted by IVM and ATV treatment. Importin and Rho GTPases gene expression was also impaired by these drugs. Furthermore, through confocal microscopy, we analyzed the effects of IVM and ATV on nuclear to cytoplasmic importin α distribution, alone or in combination. Results showed a significant inhibition of importin α nuclear accumulation under IVM and ATV treatments. These findings confirm transcriptional alterations in importins and Rho GTPases upon SARS-CoV-2 infection and point to IVM and ATV as valid drugs to impair nuclear localization of importin α when used at clinically-relevant concentrations.

Highlights

Since the early days of the coronavirus disease 2019 (COVID-19) pandemic, drug repurposing has aroused great interest in the development of novel antiviral therapies.This concept, called rediscovering or therapeutic indication expansion, implies establishing new medical uses for already known drugs [1]
We used the publicly available RNA sequencing dataset GSE152075 to evaluate the expression of the importin family genes (Importin α5 (KPNA1), Importin α1 (KPNA2), Importin α4 (KPNA3), Importin α3 (KPNA4), Importin α6 (KPNA5), Importin α7 (KPNA6), Importin α8 (KPNA7), and Importin β1 (KPNB1)) in COVID-19 positive and negative patients (Figure 1A)
When assessing gene expression in patients categorized by age, we found that age increase was significantly associated with lower KPNA2, KPNA3, KPNA4, KPNA6, and KPNB1 expression (p-trend decreasing = 0.01; p-trend decreasing = 0.004; p-trend decreasing = 0.04, p-trend decreasing = 0.002; p-trend decreasing = 0.004 respectively) and higher KPNA7 expression in COVID-19 patients (p-trend increasing = 0.016), but not in non-COVID-19 patients (Figure 1B(I,II))

Summary

Introduction

Since the early days of the coronavirus disease 2019 (COVID-19) pandemic, drug repurposing has aroused great interest in the development of novel antiviral therapies. This concept, called rediscovering or therapeutic indication expansion, implies establishing new medical uses for already known drugs [1]. Approved drugs could be used to target intracellular mechanisms in host cells that are essential for viral replication. In this regard, nuclear transport and vesicle trafficking are key cellular functions involved in the pathogenesis of RNA viruses and during cancer development

Methods

Results

Conclusion