Abstract B023: Temporally resolved proteomics identifies nidogen-2 as a co-target in pancreatic cancer that modulates fibrosis and therapy response

Abstract Pancreatic ductal adenocarcinoma (PDAC) is highly lethal, with a five-year survival rate of only ∼13%. PDAC is characterised by progressive cancer-associated fibrosis. We and others have shown that targeting cancer fibrosis can improve chemotherapy efficacy and impair metastasis in pre-clinical models1-4. As such, we aimed to use proteomics to dissect the matrix signatures of pancreatic tumours from the highly-metastatic KPC (Pdx1-Cre; LSL-Kras G12D/+ ; LSL-Trp53 R172H/+ ) and poorly-metastatic KPflC (Pdx1-Cre; LSL-Kras G12D/+ ; Trp53 fl/+ ) mouse models, with an aim to identify new fibrotic drivers in tumour evolution and metastatic cascade of this deadly disease5. We collected pancreatic tissue from KPflC, KPC and wildtype (WT) controls at early (∼50 days), mid (∼90 days) and late-stage disease (∼200 days), enriched them for matrix proteins using ISDoT de-cellularisation and analysed them using data independent acquisition liquid chromatography-tandem mass spectrometry (DIA LC-MS/MS)5. DIA LC-MS/MS revealed an increased abundance of nidogen-2 (NID2) in KPC tumours compared to KPflC, specifically during mid-stage disease5. 3D organotypic matrices generated with NID2 CRISPR interference (CRISPRi) CAFs had reduced fibrosis, shown via second harmonic generation (SHG) imaging and Picrosirius Red/birefringence analysis5. 3D organotypic invasion assays revealed that reduction of CAF NID2 significantly impeded the invasion of cancer cells5. Subcutaneous and orthotopic co-seeding experiments using NID2 CRISPRi CAFs with cancer cells showed that NID2 inhibition impeded tumour growth and fibrosis5. Intravital (in vivo) imaging with quantum dots revealed improved vascular patency in NID2 knockdown tumours, with improved response to gemcitabine/nab-paclitaxel chemotherapy5. Strikingly, in orthotopic models, mice bearing NID2 CRISPRi tumours had reduced liver metastasis and increased median survival, revealing NID2 as a potential new stromal co-target in this aggressive disease5. References 1. Vennin et al. Science Translational Medicine 9 10.1126/scitranslmed.aai8504 (2017). 2. Vennin et al. Nature Communications 10, 3637 10.1038/s41467-019-10968-6 (2019). 3. Murphy et al. Science Advances 7, eabh0363 10.1126/sciadv.abh0363 (2021). 4. Chitty et al. Nature Cancer 10.1038/s43018-023-00614-y (2023). 5. Pereira et al. Science Advances 10.1126/sciadv.adl1197 (2024). Citation Format: Brooke A. Pereira, Shona Ritchie, Cecilia R. Chambers, Katie A. Gordon, Daniel A. Reed, Morghan C. Lucas, Astrid Magenau, Kendelle J. Murphy, Max Nobis, Victoria Tyma, Jeff Evans, Lorraine Chantrill, Benjamin L. Parker, Takako Sasaki, Alexander Swarbrick1, Jennifer P. Morton6, Marina Pajic1, Sean M. Grimmond7, Anthony J. Gill8, David Herrmann1, Thomas R. Cox1, Paul Timpson. Temporally resolved proteomics identifies nidogen-2 as a co-target in pancreatic cancer that modulates fibrosis and therapy response [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Cancer Evolution: The Dynamics of Progression and Persistence; 2025 Dec 4-6; Albuquerque, NM. Philadelphia (PA): AACR; Cancer Res 2025;85(23_Suppl):Abstract nr B023.