Temporally resolved proteomics identifies nidogen-2 as a cotarget in pancreatic cancer that modulates fibrosis and therapy response
Brooke Pereira, Shona Ritchie, Cecilia R. Chambers, Katie A. Gordon, Astrid Magenau, Kendelle J. Murphy, Max Nobis, Victoria M. Tyma, Ying Fei Liew, Morghan C. Lucas, Marjan Mojtabavi Naeini, Deborah S. Barkauskas, Diego Chacon Fajardo, Anna E. Howell, Amelia L. Parker, Sean Warren, Daniel A. Reed, Victoria Lee, Xanthe L. Metcalf, Young-Kyung Lee, Luke P. O’Regan, Jessie Zhu, Michael Trpceski, Angela R. M. Fontaine, Janett Stoehr, Romain Rouet, Xufeng Lin, Jessica L. Chitty, Sean Porazinski, Sunny Z. Wu, Elysse C. Filipe, Antonia L. Cadell, Holly Holliday, Jessica Yang, Michael Papanicolaou, Ruth J. Lyons, Anaiis Zaratzian, Michael Tayao, Andrew Da Silva, Claire Vennin, Julia Yin, Alysha Dew, Paul J. McMillan, Leonard D. Goldstein, Ira W. Deveson, David R. Croucher, Michael S. Samuel, Hao‐Wen Sim, Marcel Batten, Lorraine A. Chantrill, Sean M. Grimmond, Anthony J. Gill, Jaswinder S. Samra, T.R. Jeffry Evans, Takako Sasaki, Tri Giang Phan, Alexander Swarbrick, Owen J. Sansom, Jennifer P. Morton, Australian Pancreatic Cancer Matrix Atlas (APMA), Australian Pancreatic Cancer Genome Initiative (APGI), Marina Pajic, Benjamin L. Parker, David Herrmann, Thomas R. Cox, Paul Timpson
Abstract
Pancreatic ductal adenocarcinoma (PDAC) is characterized by increasing fibrosis, which can enhance tumor progression and spread. Here, we undertook an unbiased temporal assessment of the matrisome of the highly metastatic KPC ( Pdx1-Cre , LSL-Kras G12D/+ , LSL-Trp53 R172H/+ ) and poorly metastatic KP fl C ( Pdx1-Cre , LSL-Kras G12D/+ , Trp53 fl/+ ) genetically engineered mouse models of pancreatic cancer using mass spectrometry proteomics. Our assessment at early-, mid-, and late-stage disease reveals an increased abundance of nidogen-2 (NID2) in the KPC model compared to KP fl C, with further validation showing that NID2 is primarily expressed by cancer-associated fibroblasts (CAFs). Using biomechanical assessments, second harmonic generation imaging, and birefringence analysis, we show that NID2 reduction by CRISPR interference (CRISPRi) in CAFs reduces stiffness and matrix remodeling in three-dimensional models, leading to impaired cancer cell invasion. Intravital imaging revealed improved vascular patency in live NID2-depleted tumors, with enhanced response to gemcitabine/Abraxane. In orthotopic models, NID2 CRISPRi tumors had less liver metastasis and increased survival, highlighting NID2 as a potential PDAC cotarget.