Supplementary Figures 1 through 8 from Autophagy Inhibition Dysregulates TBK1 Signaling and Promotes Pancreatic Inflammation

Yang, Shenghong; Imamura, Yu; Jenkins, Russell W.; Cañadas, Israel; Kitajima, Shunsuke; Aref, Amir; Brannon, Arthur; Oki, Eiji; Castoreno, Adam; Zhu, Zehua; Thai, Tran; Reibel, Jacob; Qian, Zhirong; Ogino, Shuji; Wong, Kwok K.; Baba, Hideo; Kimmelman, Alec C.; Pasca Di Magliano, Marina; Barbie, David A.

doi:10.1158/2326-6066.22537418.v1

Supplementary Figures 1 through 8 from Autophagy Inhibition Dysregulates TBK1 Signaling and Promotes Pancreatic Inflammation

journal contribution

posted on 2023-04-03, 23:02 authored by Shenghong Yang, Yu Imamura, Russell W. Jenkins, Israel Cañadas, Shunsuke Kitajima, Amir Aref, Arthur Brannon, Eiji Oki, Adam Castoreno, Zehua Zhu, Tran Thai, Jacob Reibel, Zhirong Qian, Shuji Ogino, Kwok K. Wong, Hideo Baba, Alec C. Kimmelman, Marina Pasca Di Magliano, David A. Barbie

Supplementary Figure 1. Characterization of inflammatory cell infiltrates and their recovery with time. Supplementary Figure 2. Negative feedback control of pTBK1 by autophagy. Supplementary Figure 3. pTBK1 levels correlate with underlying levels of basal autophagy. Supplementary Figure 4. Lack of total TBK1 enrichment in autophagosomes. Supplementary Figure 5. Screening of autophagy receptors that regulate selective autophagy of pTBK1. Supplementary Figure 6. MRT67307 or CYT387 treatment inhibits pathogen xenophagy. Supplementary Figure 7. CYT387 treatment inhibits IFNγ induced PD-L1 expression in 8988T cells. Supplementary Figure 8. Jurkat T cell recruitment by exogenous CCL5 or 8988TsgATG5 cells.

Funding

NCI

ACS Research Scholar

Lustarten Foundation

Uniting Against Lung Cancer

GTM Fund for Lung Cancer Research

History

ARTICLE ABSTRACT

Autophagy promotes tumor progression downstream of oncogenic KRAS, yet also restrains inflammation and dysplasia through mechanisms that remain incompletely characterized. Understanding the basis of this paradox has important implications for the optimal targeting of autophagy in cancer. Using a mouse model of cerulein-induced pancreatitis, we found that loss of autophagy by deletion of Atg5 enhanced activation of the IκB kinase (IKK)-related kinase TBK1 in vivo, associated with increased neutrophil and T-cell infiltration and PD-L1 upregulation. Consistent with this observation, pharmacologic or genetic inhibition of autophagy in pancreatic ductal adenocarcinoma cells, including suppression of the autophagy receptors NDP52 or p62, prolonged TBK1 activation and increased expression of CCL5, IL6, and several other T-cell and neutrophil chemotactic cytokines in vitro. Defective autophagy also promoted PD-L1 upregulation, which is particularly pronounced downstream of IFNγ signaling and involves JAK pathway activation. Treatment with the TBK1/IKKϵ/JAK inhibitor CYT387 (also known as momelotinib) not only inhibits autophagy, but also suppresses this feedback inflammation and reduces PD-L1 expression, limiting KRAS-driven pancreatic dysplasia. These findings could contribute to the dual role of autophagy in oncogenesis and have important consequences for its therapeutic targeting. Cancer Immunol Res; 4(6); 520–30. ©2016 AACR.