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

• Main Authors: Ameera M. Shaw, Ahmad Qasem, Saleh A. Naser
• Format: Article
• Language: English
• Published: MDPI AG 2021-08-01
• Series: International Journal of Molecular Sciences
• Subjects: Crohn’s Disease; Rheumatoid Arthritis; PTPN2; PTPN22; spermidine; polyamines
• Online Access: https://www.mdpi.com/1422-0067/22/16/8883

Crohn’s Disease (CD) and Rheumatoid Arthritis (RA) share some single nucleotide polymorphisms (SNPs) in protein tyrosine phosphatase non-receptor types 2 and 22 (<i>PTPN2/22</i>). Recently, we reported that clinical samples from CD and RA patients associated with <i>PTPN2:rs478582</i> or <i>PTPN22:rs2476601</i> 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 <i>PTPN22/22</i> 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 <i>PTPN2</i> SNP-edited cells, <i>PTPN2</i> expression decreased by 3.2-fold, and proliferation increased by 10.2-fold compared to control. Likewise, expression of <i>PTPN22</i> decreased by 2.4-fold and proliferation increased by 8.4-fold in <i>PTPN22</i> SNP-edited cells. IFN-γ and TNF-α secretions increased in both edited cell lines. CD25 expression (cell activation) was 80.32% in <i>PTPN2</i> SNP-edited cells and 85.82% in <i>PTPN22</i> SNP-edited cells compared to 70.48% in unedited Jurkat T-cells. Treatment of <i>PTPN2</i> and <i>PTPN22</i>-edited cells with a maximum 20 μM spermidine restored <i>PTPN2/22</i> 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 <i>PTPN2</i> or <i>PTPN22</i> 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 <i>PTPN2/22</i> 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.