MEK inhibitors reduce cellular expression of ACE2, pERK, pRb while stimulating NK-mediated cytotoxicity and attenuating inflammatory cytokines relevant to SARS-CoV-2 infection

Abstract

COVID-19 affects vulnerable populations including elderly individuals and patients with cancer. Natural Killer (NK) cells and innate-immune TRAIL suppress transformed and virally-infected cells. ACE2, and TMPRSS2 protease promote SARS-CoV-2 infectivity, while inflammatory cytokines IL-6, or G-CSF worsen COVID-19 severity. We show MEK inhibitors (MEKi) VS-6766, trametinib and selumetinib reduce ACE2 expression in human cells. In some human cells, remdesivir increases ACE2-promoter luciferase-reporter expression, ACE2 mRNA and protein, and ACE2 expression is attenuated by MEKi. In serum-deprived and stimulated cells treated with remdesivir and MEKi we observed correlations between pRB, pERK, and ACE2 expression further supporting role of proliferative state and MAPK pathway in ACE2 regulation. We show elevated cytokines in COVID-19-(+) patient plasma ( N = 9) versus control ( N = 11). TMPRSS2, inflammatory cytokines G-CSF, M-CSF, IL-1α, IL-6 and MCP-1 are suppressed by MEKi alone or with remdesivir. We observed MEKi stimulation of NK-cell killing of target-cells, without suppressing TRAIL-mediated cytotoxicity. Pseudotyped SARS-CoV-2 virus with a lentiviral core and SARS-CoV-2 D614 or G614 SPIKE (S) protein on its envelope infected human bronchial epithelial cells, small airway epithelial cells, or lung cancer cells and MEKi suppressed infectivity of the pseudovirus. We show a drug class-effect with MEKi to stimulate NK cells, inhibit inflammatory cytokines and block host-factors for SARS-CoV-2 infection leading also to suppression of SARS-CoV-2-S pseudovirus infection of human cells. MEKi may attenuate SARS-CoV-2 infection to allow immune responses and antiviral agents to control disease progression.

Identification of candidate repurposable drugs to combat COVID-19 using a signature-based approach

“…MEK inhibitors have known antiviral efficacy against coronaviruses, inhibiting the Raf/MEK/ERK signaling pathway and impairing viral production but not viral entry into the cell in a murine coronavirus hepatitis virus model 66 . It was recently shown that selumetinib, like trametinib, a MEK inhibitor also identified in this study, can stimulate natural killer cells, reduce ACE2 expression in human cells, and reduce cytokine expression in COVID-19 patient plasma, suggesting that this class of drugs may both suppress infection by SARS-CoV-2 and support the body’s immune response to infection 39 . Overall, our approach acts a useful screen for identifying common candidate drugs to reverse gene signatures for SARS-CoV-2 infected tissues despite the differences in disease transcriptome profiles from different data sources.…”

“…** Also identified in 2 different COVID-19 patient datasets. Drug Drug class Antiviral properties Trametinib Kinase inhibitor MERS-CoV 38 , SARS-CoV-2 39 Withaferin A Steroidal lactone SARS-CoV-2 40 – 43 Parthenolide Sesquiterpene lactone SARS 44 Lapatinib Kinase inhibitor SARS-CoV-2 45 Sorafenib Kinase inhibitor SARS-CoV-2 46 Auranofin Gold salt SARS-CoV-2 47 Selumetinib** Kinase inhibitor SARS-CoV-2 39 Erlotinib Antineoplastic, tyrosine kinase inhibitor HCV, RNA viruses, dengue, Ebola 48 – 50 Alvocidib CDK Inhibitor HSV, HIV, Flu 51 – 56 Quinacrine Antimalarial EMCV, poliovirus 57 Vandetanib Kinase inhibitor Andes virus 57 Dasatinib SRC tyrosine kinase inhibitor HIV 58 , 59 Thioridazine Phenothiazine …”

“…The MEK inhibitor trametibinib displayed strong inhibitory activity against MERS-CoV infection in Huh7 human hepatocytes when administered prior to and post-infection, suggesting that ERK/MAPK pathway signaling may be important for viral entry and viral replication stages of the MERS-CoV life cycle 38 . As with the candidate drug selumetinib which was one of the only drugs common to both of the patient COVID-19 sample analyses and the SARS-CoV-2 analysis, trametinib also shows efficacy against SARS-CoV-2 in vitro 39 , highlighting growing interest in the antiviral potential of this class of drug. Withaferin A has anti-inflammatory and anti-tumor properties 43 , 88 , 89 and may be useful in targeting the pathological immune component associated with COVID-19 infection.…”

Cytokine ranking via mutual information algorithm correlates cytokine profiles with presenting disease severity in patients infected with SARS-CoV-2

“…Further studies should be conducted to clarify the mechanistic role that these cytokines and macrophages play in the various stages of COVID-19 and correlate them with other hematologic parameters that were not collected in this database. The results of these future studies could identify more targeted immunomodulatory strategies beyond steroid administration such as treatment with MEK inhibitors ( Zhou et al, 2020 ), as well as the ideal timing of these interventions to maximize therapeutic efficacy. Future studies could also address the size limitations of this study, which was not powered to explore race- or ethnicity-related differences in COVID-19 severity.…”

Cytokine ranking via mutual information algorithm correlates cytokine profiles with presenting disease severity in patients with COVID-19

“…Further studies should be conducted to clarify the mechanistic role that these cytokines and macrophages play in the various stages of COVID-19. The results of these future studies could identify more targeted immunomodulatory strategies beyond steroid administration such as treatment with MEK inhibitors 13 , as well as the ideal timing of these interventions to maximize therapeutic efficacy. Finally, we present the application of this mutual information algorithm as a way to evaluate the dataset as a whole and elucidate the most important cytokines in predicting presenting severity of COVID-19.…”