Forward genetic screening in engineered colorectal cancer organoids identifies regulators of metastasis
Abstract
Metastatic outgrowth requires that cancer cells delaminate from the primary tumor, intravasate, survive in circulation, extravasate, migrate to, and proliferate at a distal site. Recurrent genetic drivers of metastasis remain elusive, suggesting that unlike the early steps of oncogenesis, metastasis drivers may be variable. We develop a framework for identifying metastasis regulators using CRISPR/Cas9-based screening in a genetically defined organoid model of colorectal adenocarcinoma. We conduct in vitro screens for invasion and migration alongside orthotopic, in vivo screens for gain of metastasis in a syngeneic mouse model. We identify CTNNA1 and BCL2L13 as bona fide metastasis-specific suppressors which do not confer any selective advantage in primary tumors. CTNNA1 loss promotes cell invasion and migration, and BCL2L13 loss promotes anchorage-independent survival and non-cell-autonomous changes to macrophage polarization. This study demonstrates proof of principle that large-scale genetic screening can be performed in tumor-organoid models in vivo and identifies novel regulators of metastasis.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (23)
Xin Wang
Zvi Cramer
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Nicolae Adrian Leu
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Keara Monaghan
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Kayla Durning
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Stephanie Adams-Tzivelekidis
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Joshua H. Rhoades
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Jonathan Heintz
Biostatistics Analysis Center, Center for Clinical Epidemiology and Biostatistics, Perelman School of Medicine, University of Pennsylvania
Yuhua Tian
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Joshua Rico
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Diego Mendez
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Ricardo Petroni
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Austin C. King
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Melissa S. Kim
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Rina Matsuda
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania
Olivia Hanselman
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Alice E. Shin
Herbert Irving Comprehensive Cancer Center, Division of Digestive and Liver, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center
María F. Carrera Rodríguez
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
Igor E. Brodsky
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania
Anil Rustgi
Herbert Irving Comprehensive Cancer Center, Division of Digestive and Liver, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center
Ning Li
Christopher J. Lengner
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania
M. Andrés Blanco