Modulation of PTPN2/22 Function by Spermidine in CRISPR-Cas9-Edited T-Cells Associated with Crohn’s Disease and Rheumatoid Arthritis

Abstract

Crohn’s Disease (CD) and Rheumatoid Arthritis (RA) share some single nucleotide polymorphisms (SNPs) in protein tyrosine phosphatase non-receptor types 2 and 22 (PTPN2/22). Recently, we reported that clinical samples from CD and RA patients associated with PTPN2:rs478582 or PTPN22:rs2476601 genotypes were linked to overactive immune response and exacerbation of inflammation. Here, we investigated in vitro the effects of these SNPs in Jurkat T-cells using CRISPR-Cas9. All cells were evaluated for PTPN22/22 loss of function and effects on cell response. We measured gene expression via RT-qPCR and cytokines by ELISA. We also measured cell proliferation using a BrdU labeling proliferation ELISA, and T-cell activation using CD-25 fluorescent immunostaining. In PTPN2 SNP-edited cells, PTPN2 expression decreased by 3.2-fold, and proliferation increased by 10.2-fold compared to control. Likewise, expression of PTPN22 decreased by 2.4-fold and proliferation increased by 8.4-fold in PTPN22 SNP-edited cells. IFN-γ and TNF-α secretions increased in both edited cell lines. CD25 expression (cell activation) was 80.32% in PTPN2 SNP-edited cells and 85.82% in PTPN22 SNP-edited cells compared to 70.48% in unedited Jurkat T-cells. Treatment of PTPN2 and PTPN22-edited cells with a maximum 20 μM spermidine restored PTPN2/22 expression and cell response including cell proliferation, activation, and cytokines secretion. Most importantly, the effect of spermidine on edited cells restored normal expression and secretion of IFN-γ and TNF-α. The data clearly demonstrated that edited SNPs in PTPN2 or PTPN22 were associated with reduced gene expression, which resulted in an increase in cell proliferation and activation and overactive immune response. The data validated our earlier observations in CD and RA clinical samples. Surprisingly, spermidine restored PTPN2/22 expression in edited Jurkat T-cells and the consequent beneficial effect on cell response and inflammation. The study supports the use of polyamines dietary supplements for management of CD and in RA patients.

Graves Disease and Inflammatory Bowel Disease: A Bidirectional Mendelian Randomization

“…This correlation implies that STAT1 may be a mediator among CD, UC, and GD. The protein tyrosine phosphatase nonreceptor type 22 (PTPN22) is one of the typical genes that share multiple variants found both in CD and GD as well as in rheumatoid arthritis; these findings are consistent with our previous MR analysis results ( 42 , 43 ).…”

Genetic Markers and Predictive Factors Influencing the Aggressive Behavior of Cerebral Cavernous Malformation

“…To date, there have been few published studies on SNPs in the PTPN2 gene, and among them, a weak association with certain diseases or different outcomes was found. However, it is important to highlight that in combination with other variants they have been shown to possibly increase the susceptibility of chronic inflammatory disorders, including rheumatoid arthritis, type 1 diabetes and celiac disease [24][25][26][27][28] . Furthermore, there have been research findings indicating the presence of epistasis between PTPN2 and VDR gene 29,30 .…”

Stem-Cell-Derived β-Like Cells with a Functional PTPN2 Knockout Display Increased Immunogenicity

“…RNAseq data suggest that the expressions of factors associated with MHC Class I regulation were upregulated in PTPN2KO sBCs, and the expressions in the NFκB signaling pathway were both up- and downregulated. It is possible that IL1 receptor levels are still intact via MAPK signaling pathways and much work has been conducted by others to understand the relationship between PTPN2 and inflammatory pathways [ 27 , 41 , 42 , 43 , 44 ]. Given that this gene expression data is not at the functional protein level, future analyses could focus on investigating the relationship between PTPN2 and these pathways in this model.…”