Remdesivir inhibits renal fibrosis in obstructed kidneys

Abstract

Aim: Kidney impairment is observed in patients with . We aimed to demonstrate the effect of anti-COVID-19 agent remdesivir on renal fibrosis.Methods: Remdesivir and its active nucleoside metabolite GS-441524 were used to treat TGF-β stimulated renal fibroblasts (NRK-49F) and human renal epithelial cells (HK2). Cell viability was determined by CCK8 assay, and fibrotic markers were measured by Western blotting. Vehicle or remdesivir were given by intraperitoneal injection or renal injection through the left ureter in unilateral ureteral obstruction (UUO) mice. Serum and kidneys were harvested. The concentrations of remdesivir and GS-441524 were measured using LC-MS/MS. Renal and liver function were assessed. Renal fibrosis was evaluated by Masson's trichrome staining and Western blotting. Results: Remdesivir and GS-441524 inhibited cell proliferation and the expression of fibrotic markers (fibronectin, pSmad3, and aSMA) in NRK-49F and HK2 cells. Intraperitoneal injection or renal injection of remdesivir attenuated renal fibrosis of UUO kidneys. Renal and liver function were not changed in remdesivir treated UUO mice. Remdesivir can not be detected, but two remdesivir metabolites were detected after injection. Conclusion: Remdesivir inhibits renal fibrosis in obstructed kidneys.was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.Remdesivir (Product name GS-5734; Cat. No. CSN19703) was purchased from CSNpharm (Chicago, Illinois, USA) and dissolved in DMSO as a 50mg/ml stock, which was further diluted into normal saline by sonication as a working solution.0.04% typan blue dye (A601140, Sangon, Shanghai, China) was added into vehicle or remdisivir working solution to monitor the injection process. 50 μ L of vehicle or remdesivir (1mg/mL) was injected retrogradely once into the left kidney via the ureter.Right after the injection, unilateral ureteral obstruction was performed. Cell cultureHK2 renal proximal tubular epithelial cells were obtained from the Cell Bank of Shanghai Institute of Biological Sciences (Chinese Academy of Science). NRK-49F rat kidney interstitial fibroblast cells were purchased from National Infrastructure of Cell Line Resource, Chinese Academy of Medical Sciences. HK2 and NRK-49F cells were cultured in DMEM/F12 medium containing 10%FBS and 0.5% penicillin and streptomycin in an atmosphere of 5% CO2 and 95% air at 37°C. For Western blotting, HK2 and NRK-49F cells were seeded in 6-well plate to 40-50% confluence, which were starved with DMEM/F12 medium 0.5% fetal bovine serum overnight before the experiment. In the next day, fresh medium containing 0.5% fetal bovine serum was changed, and then cells were exposed to 2.5 ng/ml TGF-β (Peprotech, Rocky Hill, NJ,

Clinically relevant cell culture models and their significance in isolation, pathogenesis, vaccine development, repurposing and screening of new drugs for SARS-CoV-2: a systematic review

“…293 T cell line is derived from HEK293 cell line. It is very useful for various studies such as antiviral CRISPR ( Wang et al, 2020b ), virus isolation, ( Dai et al, 2020 ), MTHFD1 as a target of anti-viral therapy ( Xing et al, 2020 ), virus susceptibility ( Thevarajan et al, 2020 ), viral pathogenesis/transmission ( Gao et al, 2020 ), vaccine production ( Mantlo et al, 2020 ), immune response ( Ye et al, 2020 ; Ju et al, 2020 ), mAb production ( Li et al, 2020 ; Sun et al, 2020 ), mAbs screening ( Fintelman-Rodrigues et al, 2020 ), vaccine production and antibodies response ( Mossel et al, 2005 ), ACE-2 gene expression in target cells ( Zhang et al, 2020a ),for drug screening like Teicoplanin ( Wu et al, 2020 ). Similarly, 293 FT cell line is used for screening of DHODH Inhibitors ( Wang et al, 2020a ), MAbs production ( Li et al, 2020 ), and 293 F used for monoclonal antibodies production ( Li et al, 2020 ; Sun et al, 2020 ); whereas, 293 T-ACE-2 cell line for vaccine production and antibodies response ( Mossel et al, 2005 ) and viral pathogenesis ( Gao et al, 2020 ).…”

“…Other respiratory systems based In Vitro Model systems are A549 cell line for ACE2 gene expression ( Zhao et al, 2020b ), and for drug screening of ATAZANAVIR ( Ge et al, 2020 ), Teicoplanin ( Wu et al, 2020 ), DHODH inhibitors ( Wang et al, 2020a ); NCI-H1299 and BEAS-2B cell lines for viral pathogenesis and transmission cell line for vaccine production ( Mantlo et al, 2020 ); Primary culture of type II alveolar (AT2) cells for ACE2 gene expression ( Poschet et al, 2020 ); IB3-1 and its derivative S9 cells for the screening of Azithromycin and Ciprofloxacin ( Wang et al, 2020c ); and KMB17 cells for vaccine production ( Mantlo et al, 2020 ). Hep-2 cell line shown significant CoVs viral titer and have ACE2 receptor ( Hoffmann et al, 2020 ; Wang et al, 2020a ), and susceptible to SARS-CoV2; however, in another study, A549 cells did not show TMPRSS2 expression, required for S protein processing and viral spread ( Matsuyama et al, 2020 ).…”

“…Based on viral load, SARS-CoVs replication in Huh-7 is comparatively higher ( Kaye, 2006 ; Fang et al, 2020 ), and suggested to be an ideal in-vitro model system. In other studies, Huh-7 has been used for Virus Isolation ( Xiao et al, 2020 ), virus susceptibility ( Thevarajan et al, 2020 ; Sungnak et al, 2020 ; Fang et al, 2020 ), Viral Pathogenesis and Transmission ( Hoffmann et al, 2020 ; Gao et al, 2020 ; Zhang et al, 2020a ; Kaye, 2006 ; Zhao et al, 2020c ), Vaccine Production ( Mantlo et al, 2020 ), gene expression ( Chan et al, 2013 ), and for drugs screening such as Teicoplanin ( Wu et al, 2020 ), and DHODH Inhibitors ( Wang et al, 2020a ).…”

Reviews on Biological Activity, Clinical Trial and Synthesis Progress of Small Molecules for the Treatment of COVID-19

“…Pruijssers et al [142] reported that Remdesivir potently inhibits SARS-CoV-2 replication in human lung cells and primary human airway epithelial cultures (EC 50 = 0.01 µM), in Vero E6 cells (EC 50 = 1.65 µM), respectively. Wu et al [143] found that Remdesivir and GS-441524 can inhibit cell proliferation and the expression of fibrotic markers (fibronectin, pSmad3, and aSMA) in NRK-49F and HK2 cells. Brandi et al [144] used rhesus macaque model of COVID-19, and showed that Remdesivir can be used in transient lower respiratory tract disease.…”