RIPK1 regulates beta-cell fate and autoimmunity in type 1 diabetes
Type 1 diabetes (T1D) results from autoimmune destruction of pancreatic beta-cells. RIPK1, a key regulator of cell death and inflammation, has been implicated in beta-cell responses to cytokines and immune attack. To investigate RIPK1's role in beta-cell fate during autoimmune stress, we generated CRISPR/Cas9-mediated Ripk1-deleted NIT-1 beta-cells and exposed them to TNFa+IFN? or self-reactive diabetic NOD splenocytes. Using RNA-seq, kinome profiling, flow cytometry, and small molecule inhibitors, we demonstrate that RIPK1 deletion or pharmacological inhibition protects beta-cells from cytokine-induced and autoimmune cell death. This study provides insights into RIPK1-dependent pathways in beta-cell vulnerability and suggests potential therapeutic strategies for T1D. Overall design: 2x2 factorial design: Genotype (Control NIT-1 vs Ripk1-deleted NIT-1) x Treatment (Vehicle vs TNFa+IFN?). NIT-1 beta-cells from NOD mice; CRISPR-engineered Ripk1? vs control cells. Cells treated with TNFa (40 ng/mL) + IFN? (100 ng/mL) or vehicle. Six biological replicates per condition (24 samples total). Multi-modal analyses: RNA-seq, kinome profiling, flow cytometry, small molecule inhibitor testing.