Abstract 3643: PTEN/STAT3 pathway in cancer-associated fibroblasts in pancreatic cancer

Abstract

Abstract This study aims to test the hypothesis that direct interactions and auto-regulatory loop between PTEN and STAT3 in cancer-associated fibroblasts (CAFs) contribute to an immunosuppressive tumor microenvironment in pancreatic ductal carcinoma (PDAC). Pancreatic cancer accounts for almost as many deaths as cases because of its poor prognosis. Many studies have led to the paradigm that stromal elements contribute to pancreatic tumor growth. Cancer-associated fibroblasts (CAFs) are a primary cell type in the desmoplastic stroma in PDAC, and targeting them to improve PDAC treatment is thought to be a promising strategy, however; studies have shown both tumor-promoting and tumor-limiting activities associated with PDAC CAFs. Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is one of the most well-described tumor suppressor genes and can regulate essential transcription factors in stromal fibroblasts. On the other hand, STAT3 signaling in pancreatic CAFs contributes to the immunosuppressive and fibrotic stroma observed with disease progression. In this study, we used novel, dual recombinase mouse models of PDAC, with Kras and p53 mutations engineered in the pancreatic epithelium (Flp-based) and Stat3 deleted in a subpopulation of CAFS (Cre). CAFs isolated from these mice were used to probe PTEN-STAT3 interaction in vitro using the Proximity Ligation Assay (PLA) and by ChIP-seq using an RNA seq. Mouse and Human pancreatic tumor samples were analyzed and quantified using multiplex-immunofluorescence and multispectral imaging. In the mouse genetic model, we found PTEN downregulated in ~80% of CAFs, consistent with previous work from our group that demonstrated PTEN loss in 25% of PDAC patient samples. At the same time, pY705-STAT3 was activated in CAFs, and deletion of STAT3 in CAFs correlated with increased survival, decreased M2-like macrophages, and increased T-cell infiltration. STAT3 ablation in CAFS resulted in the re-expression of PTEN in CAFS, implying an autoregulatory loop. PLA on CAFs isolated revealed that IL6 treatment resulted in the transfer of a STAT3-PTEN protein complex into the nucleus ChIP-seq on CAFs with PTEN STAT3 antibodies revealed a significant overlap in PTEN and STAT3 occupation of gene promoters. This result is consistent with in silico analysis on the PTEN and STAT3 ChIP-Seq data in human cell lines. ChIP-Seq data is being interacted with RNA data to identify essential target genes. Human PDAC tissue microarrays are stained with pSTAT3, PTEN, CD3, and CD8.Our study suggests an autoregulatory look between PTEN and STAT3 in PDAC CAFs. PTEN may suppress specific STAT3 target genes through direct interactions with chromatin. Alternatively, PTEN/STAT3 complexes may target CAF genes that suppress tumor growth or activate anti-tumor immune responses. These investigations could identify potential targets in CAFs for restoring the anti-tumor immune responses in PDAC. Citation Format: Samaneh Saberi, Sudarshana Sharma, Cameron Bumbleburg, Julia E. Lefler, Michael C. Ostrowski. PTEN/STAT3 pathway in cancer-associated fibroblasts in pancreatic cancer. [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 3643.