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Supplementary Figures from Small-Molecule Inhibition of Axl Targets Tumor Immune Suppression and Enhances Chemotherapy in Pancreatic Cancer

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posted on 2023-03-31, 01:21 authored by Kathleen F. Ludwig, Wenting Du, Noah B. Sorrelle, Katarzyna Wnuk-Lipinska, Mary Topalovski, Jason E. Toombs, Victoria H. Cruz, Shinichi Yabuuchi, N.V. Rajeshkumar, Anirban Maitra, James B. Lorens, Rolf A. Brekken

Supplementary Figures 1-7 and Supplementary Table 1. Supplementary Figure 1 H&E stain and trichrome stain of representative 7-week-old KIC tumor tissue. Supplementary Figure 2 Axl gene expression in pancreatic cancer patient-derived xenografts. Supplementary Figure 3 Immunofluorescence of cleaved-caspase 3, p-histon-3, endomucin and EMT markers in Pan02 tumor tissue. Supplementary Figure 4 Western blot of TBK1/NF-κB signaling in tumor lysates from KIC mice treated with/without BGB324. Supplementary Figure 5 F4/80 stain in tumor tissue from KIC mice treated with BGB324 +/- gemcitabine. Supplementary Figure 6 Arginase 1 mRNA expression level in bone marrow-derived macrophages treated with IL-4 +/- BGB324. Supplementary Figure 7 Flow cytometry of KPC-M09 subcutaneous tumors treated with/without BGB324. Supplementary Table 1 Cytokine/Chemokine Concentration in KIC tumor lysates from different treatment groups.

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ARTICLE ABSTRACT

Activation of the receptor tyrosine kinase Axl is associated with poor outcomes in pancreatic cancer (PDAC), where it coordinately mediates immune evasion and drug resistance. Here, we demonstrate that the selective Axl kinase inhibitor BGB324 targets the tumor–immune interface to blunt the aggressive traits of PDAC cells in vitro and enhance gemcitibine efficacy in vivo. Axl signaling stimulates the TBK1–NFκB pathway and innate immune suppression in the tumor microenvironment. In tumor cells, BGB324 treatment drove epithelial differentiation, expression of nucleoside transporters affecting gemcitabine response, and an immune stimulatory microenvironment. Our results establish a preclinical mechanistic rationale for the clinical development of Axl inhibitors to improve the treatment of PDAC patients.Significance: These results establish a preclinical mechanistic rationale for the clinical development of AXL inhibitors to improve the treatment of PDAC patients. Cancer Res; 78(1); 246–55. ©2017 AACR.