Abstract
Abstract
Background: Pancreatic ductal adenocarcinoma (PDAC) remains a major therapeutic challenge with hallmarks including oncogenic mutations, desmoplastic architecture, and an immunosuppressive tumor microenvironment (TME). Previously, we have identified Cyclic AMP Response Element Binding protein 1 (CREB) as an oncogenic transcriptional factor downstream of KRAS that promotes disease aggressiveness, poor survival, and immune exclusion. Based on these, we sought to determine the impact of tumor intrinsic CREB deletion in shaping the tumor immune microenvironment.
Methods: We have generated a novel genetically engineered mouse model (GEMM) of pancreas-specific CREB deletion (CREBfl/fl) in LSL-KrasG12D/+; Trp53 R172H/+; Pdx1Cre/+ (KPC) mice that phenocopy human PDAC disease. CRISPR/CAS9-based genomic editing was utilized to ablate CREB (CREBKO) in KPC tumor cells. RNA-sequencing analysis was performed in KPC CREB wild type (CREBEV) vs. CREBKO tumor cells to identify CREB-mediated transcriptomic changes. Syngeneic orthotopic tumor implantation of these cells was performed into the pancreata of mice. High dimensional immunophenotyping was accomplished to assess changes in the immune subsets with CREB deletion in murine tumors. Additionally, these tissues were processed for immunohistochemical and qPCR-based analysis to assess changes in fibroinflammatory and immune mediators.
Results: Pancreas-specific CREB deletion in the KPC GEMM led to a significant reduction in the primary tumor burden, liver metastases and improved overall survival compared to wild-type KPC. Additionally, CREB deletion significantly remodeled the tumor stroma, as evidenced by the reduction in the expression of fibroinflammatory and immunosuppressive markers. In assessing the immune repercussions of CREB deletion in pancreatic tumors, we observed a decreased infiltration of CD11b+ myeloid-derived suppressor cells (MDSCs) and granulocytic-PMN MDSCs (CD11b+ Ly6Ghigh Ly6Clow F4/80-), with a concomitant increase in the antigen-presenting M1-like macrophages (F4/80+ MHC-II high CD86 high). Also, CREB ablation in these tumors further facilitated increased infiltration of activated effector CD8+ T cells resulted in enhanced anti-tumor immune response within the PDAC TME. Mechanistically, RNA transcriptomic and secretome analysis in CREBKO Vs. the wild type KPC tumor cells identified several differentially expressed immunomodulatory soluble mediators responsible for shaping CREB dependent immunogenic landscape in PDAC.
Conclusion: Overall, depleting CREB reshapes the tumor immune landscape to reduce innate immunosuppressive myeloid infiltration and reinvigorate the antitumor T cell immune responses to improve overall survival in PDAC.
Citation Format: Siddharth Mehra, Vanessa Garrido, Samara Singh, Iago D Silva, Luis Alberto Nivelo, Anna Bianchi, Shrey Modi, Zhiqun Zhou, Austin Dosch, Nilesh Deshpande, Supriya Srinivasan, Christine Rafie, Ifeanyichukwu Ogobuiro, Xi Chen, Alejandro Villarino, Jashodeep Datta, Nipun Merchant, Nagaraj Nagathihalli. Deciphering the role of CREB1 in shaping the tumor immune landscape of pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1328.