Abstract 1444: Modeling Twist1 overexpression in a pancreatic ductal adenocarcinoma mouse model

Abstract

Abstract Pancreatic ductal adenocarcinoma (PDAC) is an extremely lethal cancer characterized by very poor survival outcomes. Genomic sequencing of human PDAC tissues and cell lines have identified four molecular subtypes: quasimesenchymal (QM), pancreatic progenitor, immunogenic, and aberrantly differentiated endocrine exocrine (ADEX). QM-PDAC has the worse prognosis, but no in vivo models exist for this molecular subtype. Our preliminary analysis using publicly available data demonstrated that the epithelial-to-mesenchymal transition (EMT) transcription factor TWIST1 is upregulated in QM-PDAC. We hypothesized that the TWIST1-dependent plasticity program was a key regulator of QM-PDAC subtype development. We created a tetracycline-inducible tissue specific Twist1 PDAC genetically engineered mouse model (GEMM). Tumor development in the mouse pancreatic ductal epithelium was directed using the Pdx1 promoter-Cre (P) allele leading to stable native expression of the LSL-KrasG12D oncogene (R) and LSL-rtTA-IRES-GFP (G) alleles. The non-Twist1 GEMM (PGR) was crossed with the Twist1-tetO-Luc (T) mouse to create the PGRT model which allows inducible expression of Twist1 and luciferase in pancreatic epithelium under doxycycline treatment (Tet-ON system). Tumor progression, lineage tracing and metastatic invasion are followed by fluorescence (GFP) and bioluminescence (BLI) imaging. The characterization of this novel PGRT mouse model showed decreased overall survival compared to control non-Twist1 containing mice (~35 weeks PGRT (n=17) vs ~60 weeks PGR (n=10)). Histological and immunohistochemistry analyses showed the PGRT pancreatic tumors resemble the QM-PDAC subtype and vimentin overexpression correlates with Twist1-dependent induction of EMT. We attempted to confirm the QM-PDAC profile via cellular markers: GATA6-, HNF1A-, KRT81+, KRT17+, KRT20+, P63+ and ∆P63+. The PGR and standard KPC (Kras+/LSL-G12D; Trp53+LSL-R172H; Pdx1-Cre) PDAC model were also profiled. We observed that 75% of the PGRT tumors displayed a QM-PDAC profile, contrasting with the absence of such profile in both PGR and KPC tumors. Examining the tumors from PGR and KPC, 100% of the PGR and 72.27% KPC tumors exhibited a non-squamous PDAC profile while an intermediate squamous profile was observed in 25% PGRT tumors and 27.27% KPC tumors. Furthermore, Twist1 induction resulted in intra-abdominal and lung metastases. We observed that 87.5% of the PGRT mice were positive for metastases while we observed no positivity for PGR mice and 63.64% positivity in KPC mice. In conclusion, these data suggest that Twist1 overexpression cooperates with KrasG12D for PDAC tumor development mimicking the human QM-PDAC subtype to promote progression towards metastasis and early lethality. Potential uses of this novel model include testing treatment strategies to ultimately improve patient outcomes with QM-PDAC. Citation Format: Muhammad Ajmal Khan, Jinhee Chang, Triet Nguyen, Danielle N. Council, Dipanwita Dutta Chowdhury, Amol Shetty, Yang Song, Aaron Chan, Christine Lam, Priyanshi Patel, Lucas Tran, Jiayu Chen, Katriana Nugent, Matthew Ballew, Ghali Lemtiri-Chlieh, Francesca A. Carrieri, Reem Malek, Hailun Wang, Noelle R. Thielman, Keyu Li, Lei Zheng, Kathleen Gabrielson, Phuoc T. Tran, Audrey Lafargue. Modeling Twist1 overexpression in a pancreatic ductal adenocarcinoma mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1444.