Abstract 3627: Inhibition of tumor cell-autonomous p38 MAPK suppresses IL1α-mediated inflammatory tumor-stromal crosstalk in pancreatic adenocarcinoma
Authors: Singh Samara , Dosch Austin R. , Mehra Siddharth , Silva Iago de Castro , Bianchi Anna , Garrido Vanessa Tonin , Zhou Zhiqun , Amirian Haleh , Box Edmond W. , Datta Jashodeep , Nagathihalli Nagaraj , Merchant Nipun B. 
Published: April 2023

Abstract
Introduction: Pancreatic ductal adenocarcinoma (PDAC) continues to face major therapeutic challenges due to its innate and acquired chemoresistance. Oncogenic, activating KRAS mutations facilitate extensive cytokine-mediated crosstalk between tumor cells and the fibrotic stroma, producing a pro-inflammatory and immunosuppressive tumor microenvironment (TME). We have identified interleukin-1α (IL1α), downstream of KRAS, as a critical mediator of the inflammatory response due to its pleiotropic effects on cancer-associated fibroblast (CAF) activation and immune evasion. However, the regulatory mechanisms of IL1α expression remain incompletely understood. We identified p38 stress-associated MAPK α (p38α) as a central, KRAS-driven regulatory pathway involved in IL1α production within PDAC tumor cells.
Methods: Differential gene expression analyses were performed on patient data from The Cancer Genome Atlas (TCGA) was queried for overactive pathways downstream KRAS in “high-IL1A” PDAC cases. Inhibition of p38α was achieved pharmacologically with pexmetinib and genetically with an shRNA lentiviral system in human and murine PDAC cell lines. ChIP-qPCR and co-immunoprecipitation were performed on a human PDAC cell line with and without p38α inhibition. Single-cell RNA sequencing (scRNA-seq) and flow cytometry were performed on tumors from a PDAC murine model, Ptf1aCre/+; LSL-KrasG12D/+; Tgfbr2flox/flox (PKT), treated with or without pexmetinib. PKT mice were treated daily with pexmetinib, gemcitabine and paclitaxel chemotherapy, or combination therapy for downstream analysis and survival studies.
Results: Both pharmacologic and genetic inhibition of p38α significantly reduced IL1A transcription and protein levels in human and murine PDAC tumor cell lines. Furthermore, p38α inhibition reduced binding of Sp1 and NF-κB to the IL1A promoter and prevented IL1α-mediated polarization of inflammatory CAFs in vitro. In PKT mice, p38α inhibition reduced tumor cell-specific Il1a and inflammatory CAF gene signatures by scRNA-seq, and favorably altered the infiltrating immune populations by flow cytometry. Lastly, pexmetinib with chemotherapy significantly reduced tumor burden and improved overall survival in a PDAC murine model.
Conclusions: These findings provide a new therapeutic opportunity to target the p38α MAPK pathway for suppression of IL1α-mediated stromal activation and combination with chemotherapy to overcome therapeutic resistance by modulating the stromal and immune microenvironment in PDAC.
Citation Format: Samara Singh, Austin R. Dosch, Siddharth Mehra, Iago de Castro Silva, Anna Bianchi, Vanessa Tonin Garrido, Zhiqun Zhou, Haleh Amirian, Edmond W. Box, Jashodeep Datta, Nagaraj Nagathihalli, Nipun B. Merchant. Inhibition of tumor cell-autonomous p38 MAPK suppresses IL1α-mediated inflammatory tumor-stromal crosstalk in pancreatic adenocarcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3627.