Fig. 2

IL1RAP knockdown and IRAK4 inhibition reduce pancreatic cancer cell growth. A, B: siRNA mediated knockdown of IL1RAP leads to Go/G1 cell cycle arrest in A6L PDAC cells. Representative flow plots are shown. Means ± stdev, N = 10 (B). Ttest P Val < 0.01. C: RNAseq done on A6L PDAC cells with and without siRNA mediated knockdown of IL1RAP show reduction in numerous genes. D, E: Immunoblotting shows reduced levels of proliferative phosphor/activated erk MAPK kinase (D) and CDK1 and Topoisomerase 2a (E) in cells with IL1RAP knockdown. F: IRAK4 is a targetable kinase downstream of IL1RAP activation. CA-4948 and PF06650833 are clinically active small molecule specific inhibitors of IRAK4. G: Pharmacologic inhibition of IRAK4 with CA4948 and PF06650833 leads to reduced viability in Panc1 and A6L PDAC cells. Means ± stdev, N = 3, Ttest *P Val < 0.05. H, I: Immunblotting shows reduced levels of proliferative phospho/activated erk MAPK kinase after IRAK4 inhibition in A6L PDAC cells. J, K: PDAC xenografts were established with subcutaneous A6L implantation in NSG mice. Treatment with 50 mg/kg/d of CA4948 (J) and PF06650833 (K) with oral gavage was initiated and tumor sizes were measured. Significant reduction in tumor volumes was observed with inhibition of IRAK4 in vivo. (Means ± s.e.m, N = 28 for CA4948; N = 10 for PF06650833). *P Val < 0.05)