Effects of immunophilin inhibitors and non-immunosuppressive analogs on coronavirus replication in human infection models

Abstract

RationaleHuman coronaviruses (HCoVs) seriously affect human health by causing respiratory diseases ranging from common colds to severe acute respiratory diseases. Immunophilins, including peptidyl-prolyl isomerases of the FK506-binding protein (FKBP) and the cyclophilin family, are promising targets for pharmaceutical inhibition of coronavirus replication, but cell-type specific effects have not been elucidated. FKBPs and cyclophilins bind the immunosuppressive drugs FK506 and cyclosporine A (CsA), respectively.MethodsPrimary human bronchial epithelial cells (phBECs) were treated with CsA, Alisporivir (ALV), FK506, and FK506-derived non-immunosuppressive analogs and infected with HCoV-229E. RNA and protein were assessed by RT-qPCR and immunoblot analysis. Treatment with the same compounds was performed in hepatoma cells (Huh-7.5) infected with HCoV-229E expressing Renilla luciferase (HCoV-229E-RLuc) and the kidney cell line HEK293 transfected with a SARS-CoV-1 replicon expressing Renilla luciferase (SARS-CoV-1-RLuc), followed by quantification of luminescence as a measure of viral replication.ResultsBoth CsA and ALV robustly inhibited viral replication in all models; both compounds decreased HCoV-229E RNA in phBECs and reduced luminescence in HCoV-229E-RLuc-infected Huh7.5 and SARS-CoV-1-RLuc replicon-transfected HEK293. In contrast, FK506 showed inconsistent and less pronounced effects in phBECs while strongly affecting coronavirus replication in Huh-7.5 and HEK293. Two non-immunosuppressive FK506 analogs had no antiviral effect in any infection model.ConclusionThe immunophilin inhibitors CsA and ALV display robust anti-coronaviral properties in multiple infection models, including phBECs, reflecting a primary site of HCoV infection. In contrast, FK506 displayed cell-type specific effects, strongly affecting CoV replication in Huh7.5 and HEK293, but inconsistently and less pronounced in phBECs.

Quantitative proteomics of differentiated primary bronchial epithelial cells from chronic obstructive pulmonary disease and control identifies potential novel host factors post-influenza A virus infection

“…The primary human bronchial epithelial cells were expanded and cultured as described previously ( Berthold et al, 2022 ; Mastalerz et al, 2022 ). For more details, refer to the online Supplementary material .…”

“…Primary human bronchial epithelial cells (phBECs) differentiated at the air–liquid interface (ALI) represent a powerful tool to assess the airway-specific first-line defense against external stimuli. The resulting organotypic bronchoepithelium closely resembles the in vivo airway and is frequently used in studies of toxicology, infection, and drug delivery ( Pezzulo et al, 2011 ; Lenz et al, 2014 ; Singanayagam et al, 2019 ; Berthold et al, 2022 ; Mastalerz et al, 2022 ). The fully differentiated epithelium, obtained 4 weeks after the airlift, contains all major bronchial epithelial cell types and displays physiologically important functional properties like mucus secretion and ciliary beating ( Lodes et al, 2020 ).…”

SARS‐CoV‐2, periodontal pathogens, and host factors: The trinity of oral post‐acute sequelae of COVID‐19

“…273 Inhibition of calcineurin blocks NF-AT transduction, subsequently inhibiting IL-2 production and the activation of T-cells. 256 Blocking NF-AT impairs adept activation of pro-inflammatory cytokines (IL-2, IL-4, IL-6, and IL-8), resulting in a weaker inflammatory response. 274 Other immunosuppressive functions of FKBPs include indirectly regulating AHR transcription and complexing with immunosuppressive drugs FKBP12 and the mTOR inhibitor rapamycin.…”

The impact of COVID-19 on liver transplantation: challenges and perspectives

“…It is also hypothesized that drugs such as cyclosporine and tacrolimus could inhibit viral replication by suppression of the immunophilin pathway. 9 Mycophenolate mofetil, a key component in the first 6–12 months of post-transplant immunosuppression, has also been shown to inhibit SARS-CoV-2 replication in vitro . 10 This probably leads to a diminished response and a less severe clinical course in these patients.…”