SARS-CoV-2 attenuates corticosteroid sensitivity by suppressing DUSP1 expression and activating p38 MAPK pathway

Abstract

The efficacy of corticosteroids and its use for the treatment of SARS-CoV-2 infections is controversial. In this study, using data sets of SARS-CoV-2 infected lung tissues and nasopharyngeal swabs, as well as in vitro experiments, we show that SARS-CoV-2 infection significantly downregulates DUSP1 expression. This downregulation of DUSP1 could be the mechanism regulating the enhanced activation of MAPK pathway as well as the reported steroid resistance in SARS-CoV-2 infection. Moreover, chloroquine, an off labeled COVID-19 drug is able to induce DUSP1 and attenuate MAPK pathway; and is expected to improve sensitivity to steroid treatment. However, further mechanistic studies are required to confirm this effect.

Keywords: COVID-19; Chloroquine; Corticosteroid; DUSP1; SARS-CoV-2; p38 MAPK pathway.

Fig. 1

SARS-CoV-2 attenuated corticosteroid sensitivity by suppressing DUSP1 Expression. (A) Log fold change (LogFC) expression of glucocorticoid-dependent genes in SARS-CoV-2 (n = 3 infected human airway epithelial cells [HAECs] vs. n = 3 mock-treated HAECs; GSE147507) or SARS-CoV-1 (n = 11 infected HAECs vs. n = 9 mock-treated HAECs; GSE47960, GSE47961, and GSE47962) infected HAECs as well as lung autopsies of COVID-19 patients (n = 17 SARS-CoV-2 infected lung vs. n = 5 healthy lung biopsies; GSE150316). The data presented shows significantly lower expression levels of DUSP1 and TSC22D3 (GILZ) in SARS-CoV-2 compared to SARS-CoV-1 infections (2 LogFC decrease with DUSP1, P = 0.0003; and 0.88 LogFC decrease with TSC22D3, P = 0.032, with SARS-CoV-2 vs SARS-CoV-1 infection, respectively). (B and C) Gene expression levels of DUSP1 and TSC22D3 (GILZ) in COVID-19 (n = 430) and in healthy (n = 54) nasopharyngeal swabs (GSE152075). The data shows expression levels of these genes are significantly lower in nasopharyngeal swabs from COVID-19 patients as compared to healthy controls. (D) Confirmation of gene expression levels of DUSP1 and TSC22D3 (GILZ), as measured by RT-qPCR, in severe COVID-19 nasopharyngeal swabs (n = 36) and in healthy nasopharyngeal swabs (n = 6). The data shows expression levels of these genes are lower in COVID-19 nasopharyngeal swabs compared to healthy nasopharyngeal swabs. (E) Protein expression of DUSP1 in nasopharyngeal swabs from severe COVID-19 patients (n = 5) and in healthy nasopharyngeal swabs (n = 4) that were randomly chosen from the 36 patient cohorts. The data presented shows reduced protein expression of DUSP1 in COVID-19 nasopharyngeal swabs compared to healthy nasopharyngeal swabs. Statistical test: LogFC was determined using adjusted LIMMA and two-way comparison was done using unpaired t-test or Mann-Whitney U test, depending on the skewness of the data. ns = non-significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Fig. 2

Increased MAPK signaling and proinflammatory genes production following SARS-CoV-2 infection. (A) Enrichment of MAPK pathway in SARS-CoV-2 infected HAECs. (Enrichment Score (ES): 0.437, Normalized enrichment Score (NES): 1.57). (B) Enrichment of MAPK pathway in SARS-CoV-1 infected HAECs. (C) LogFC expression of proinflammatory genes in SARS-CoV-2 (n = 3 infected HAECs vs. n = 3 mock-treated HAECs; GSE147507) or SARS-CoV-1 (n = 11 infected HAECs vs. n = 9 mock-treated HAECs; GSE47960, GSE47961, and GSE47962) infected HAECs as well as lung autopsies of COVID-19 patients (n = 17 SARS-CoV-2 infected lung vs. n = 5 healthy lung biopsies; GSE150316). The data presented shows higher level of proinflammatory genes expression in SARS-CoV-2. (E) Protein expression of p-ERK 1/2 in nasopharyngeal swabs from severe COVID-19 patients (n = 5) and in healthy nasopharyngeal swabs (n = 4) that were randomly chosen from the 36 patient cohorts (Fig. 1D). The data presented shows increased expression of p-ERK 1/2 in COVID-19 nasopharyngeal swabs as compared to healthy nasopharyngeal swabs; indicative of increased MAPK signaling. Statistical test: LogFC was determined using adjusted LIMMA and two-way comparison was done using unpaired t-test or Mann-Whitney U test, depending on the skewness of the data. ns = non-significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Fig. 3

Chloroquine attenuated MAPK signaling modulating steroid resistance. (A) The level of DUSP1 expression in chloroquine treated hepatocytes dataset (n = 6 treated hepatocytes vs. n = 3 untreated hepatocytes; GSE30351). The data presented shows elevated level of DUSP1 expression in chloroquine treated-compared to untreated hepatocytes. (B) Enrichment of MAPK pathway in chloroquine-treated hepatocytes. (C) Expression level of p-ERK 1/2 in chloroquine or/and dexamethasone treated bronchial fibroblasts of COPD. The data presented shows reduced level of p-ERK 1/2 expression in chloroquine treated as well as in chloroquine and dexamethasone treated compared to untreated bronchial fibroblasts of COPD; indicative of less MAPK activation. (D) Expression level of IL-1β in chloroquine or/and dexamethasone treated bronchial fibroblasts of COPD. The data presented shows reduced level of this cytokine in chloroquine treated as well as in chloroquine and dexamethasone treated fibroblasts compared to untreated fibroblasts. Two-way comparison was done using unpaired t-test or Mann-Whitney U test, depending on the skewness of the data. *P < 0.05, **P < 0.01.