Inflammatory autoimmune diseases like Crohn's Disease (CD) and Rheumatoid Arthritis (RA) share some of the same single nucleotide polymorphisms (SNPs) in protein tyrosine phosphatase non-receptor types 2 and 22 (PTPN2/22), which contribute to their pathogenesis. In clinical CD and RA samples, PTPN2:rs478582 and PTPN22:rs2476601 were found to exacerbate a number of inflammatory processes associated with CD and RA. To confirm the role of these SNPs in CD and RA pathogenesis, CRISPR-Cas9 was used to induce the SNPs in T-cells. Cells were also treated with the naturally occurring polyamine, spermidine, to restore PTPN2/22 function and reverse the inflammatory effects of the SNPs. The PTPN2 SNP decreased PTPN2 expression by 3.2-fold and the PTPN22 SNP decreased PTPN22 expression by 2.4-fold compared to WT T-cells. Proliferation increased by 10.2-fold in PTPN2 SNP cells and 8.4-fold in the PTPN22 SNP cells compared to WT cells. Both SNPs increased secretion levels IFN-γ and TNF-α. Additionally, 80.32% of PTPN2 SNP cells and 85.82% of PTPN22 SNP cells were activated compared to 70.48% of WT T-cells. Spermidine treatment increased PTPN2/22 expression in all cell types and was dose-dependent in cells with either SNP. Proliferation levels decreased by approximately 5-fold in spermidine treated PTPN2 SNP cells and 3.1-fold in PTPN22 SNP cells treated with 10 µM of spermidine and 12.0-fold when treated with 20 uM. IFN-γ and TNF-α secretion levels decreased with spermidine treatment in all cell groups. Lastly, T-cell activation decreased to 51.39% of PTPN2 SNP cells and 46.36% of PTPN22 SNP cells when treated with 10 µM of spermidine. These findings may explain the therapeutic response seen in CD and RA patients prescribed polyamines. This study confirms the inflammatory role of the PTPN2/22 SNPs and displays the anti-inflammatory and restorative property of spermidine for its potential therapeutic use in CD and RA patients with genetic polymorphisms.
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Master of Science (M.S.)
College of Medicine
Burnett School of Biomedical Sciences
Length of Campus-only Access
Masters Thesis (Open Access)
Shaw, Ameera, "Spermidine Rescued PTPN2/22 Function in CRISPR-Cas9-edited T-cells with PTPN2/22 SNPs Linked to Crohn's Disease and Rheumatoid Arthritis" (2021). Electronic Theses and Dissertations, 2020-. 1158.