The JAK-Inhibitor Tofacitinib Rescues Human Intestinal Epithelial Cells and Colonoids from Cytokine-Induced Barrier Dysfunction

Abstract

Background Alterations to epithelial tight junctions can compromise the ability of the epithelium to act as a barrier between luminal contents and the underlying tissues, thereby increasing intestinal permeability, an early critical event in inflammatory bowel disease (IBD). Tofacitinib (Xeljanz), an orally administered pan-Janus kinase (JAK) inhibitor, was recently approved for the treatment of moderate to severe ulcerative colitis. Nevertheless, the effects of tofacitinib on intestinal epithelial cell functions are largely unknown. The aim of this study was to determine if JAK inhibition by tofacitinib can rescue cytokine-induced barrier dysfunction in intestinal epithelial cells (IECs). Methods T84 IECs were used to evaluate the effects of tofacitinib on JAK-signal transducer and activator of transcription (STAT) activation, barrier permeability, and expression and localization of tight junction proteins. The impact of tofacitinib on claudin-2 promoter activity was assessed in HT-29 IECs. Tofacitinib rescue of barrier function was also tested in human colonic stem cell-derived organoids. Results Pretreatment with tofacitinib prevented IFN-γ-induced decreases in transepithelial electrical resistance (TER) and increases in 4 kDa FITC-dextran permeability (FD4), partly due to claudin-2 transcriptional regulation and restriction of ZO-1 rearrangement at tight junctions. Although tofacitinib administered after IFN-γ challenge only partially normalized TER and claudin-2 levels, FD4 permeability and ZO-1 localization were fully recovered. The IFN-γ-induced FD4 permeability in primary human colonoids was fully rescued by tofacitinib. Conclusions These data suggest differential therapeutic efficacy of tofacitinib in the rescue of pore vs leak-tight junction barrier defects and indicate a potential contribution of improved epithelial barrier function to the beneficial effects of tofacitinib in IBD patients.

Patient Derived Colonoids as Drug Testing Platforms–Critical Importance of Oxygen Concentration

“…We have previously confirmed that organoids developed from human colonic tissue (colonoids) contain stem cells and all differentiated cell lineages present in the colonic epithelium, and develop a 3D structure with budding crypts and surface areas encircling the lumen ( Ostvik et al, 2020 ). Thus, from our results and those from others ( Dotti et al, 2017 ; Arnauts et al, 2020 ; Sayoc-Becerra et al, 2020 ) colonoids emerge as a putative tool to study effects of existing or emerging IBD treatments targeting epithelial cells even in a specific patient.…”

HO‐1/CO Maintains Intestinal Barrier Integrity through NF‐κB/MLCK Pathway in Intestinal HO‐1-/- Mice

“…Our study further verified the clinical significance of HO-1/CO repairing intestinal barrier injury by alleviating hepatic fibrogenesis and serum ALT levels. Intestinal epithelial barrier disruption can enhance or directly trigger IBD or dysbiosis [ 40 , 41 ]; therefore, it is of great significance to study the effects of HO-1/CO maintaining intestinal barrier integrity on targeted treatment of IBD or intestinal microecological diseases.…”

Identification of a Novel Susceptibility Marker for SARS-CoV-2 Infection in Human Subjects and Risk Mitigation with a Clinically Approved JAK Inhibitor in Human/Mouse Cells

“…For PTPN2 knockdown, the cells were infected with lentiviral particles containing non-targeting control shRNA (Ctr) or PTPN2-specfic shRNA as described previously 49 and stable clones selected using puromycin. For STAT1 silencing, the cells were transfected with previously validated, STAT1-specific or non-targeting control siRNA constructs (Dharmacon) using DharmaFECT transfectionre agents as described previously 50 . In experiments with STAT1 siRNA and IFN-γ treatment, the culture medium was replaced with serum-free medium 8 h prior to addition of IFN-γ (1000 IU; Roche).…”