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

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.

Safety and Efficacy of a MEURI Program for the Use of High Dose Ivermectin in COVID-19 Patients

“…The role of IVM against SARS-CoV-2 was supported by the biologic plausibility based on in vitro and in vivo studies of mechanistic analyses and antiviral effects. Its proposed antiviral mechanism is thought to be mediated by its ability to inhibit the nuclear import of viral proteins mediated by IMPa/b1 heterodimer, and the promotion of defense mechanisms such as pyroptosis in infected epithelial cells, suggesting its possible role as a broad spectrum antiviral agent ( 30 , 31 ). Although an immunomodulatory effect of IVM has been proposed by other authors ( 32 ), these effects might explain the antiviral activity against SARS-CoV-2 reported by our group and Biber et al, in two small randomized controlled trials ( 17 , 19 ).…”

Antiviral Activity of Repurposing Ivermectin against a Panel of 30 Clinical SARS-CoV-2 Strains Belonging to 14 Variants

“…The in vitro combination of ivermectin (2 µM) and remdesivir (6 µM) shows highly synergistic effects against the murine hepatitis virus (MHV), which belongs to the betacoronavirus genus like the SARS-CoV-2 [25]. Ivermectin exerts higher in vitro inhibition of importin α nuclear accumulation in combination with atorvastatin than when used alone [84]. A randomised, blind trial in patients with mild-to-moderate COVID-19 symptoms showed that patients treated with ivermectin 12 mg and doxycycline 100 mg, twice a day for five days, recovered earlier than those receiving standard care alone (paracetamol, antihistaminics, vitamins, low molecular weight heparin and oxygen therapy if necessary), and were more significantly asymptomatic after 12 days and were less likely to be diagnosed with SARS-CoV-2 after 14 days [85].…”

Pin-Pointing the Key Hubs in the IFN-γ Pathway Responding to SARS-CoV-2 Infection

“…Briefly, A549 cells were plated in 6-well flat bottom plates at a density of 2 × 10 5 cells in complete DMEM, allowed to attach overnight, and then transfected with Poly (I:C) or distilled water as mock in DMEM without FBS and antibiotics. After 6 h, the media were replaced, and 24 h later, total cell lysates and RNA isolation was carried out [ 36 ].…”