Forward genetic screening in engineered colorectal cancer organoids identifies regulators of metastasis
- Xin Wang
- Zvi Cramer
- Nicolae Adrian Leu
- Keara Monaghan
- Kayla Durning
- Stephanie Adams-Tzivelekidis
- Joshua H. Rhoades
- Jonathan Heintz
- Yuhua Tian
- Joshua Rico
- Diego Mendez
- Ricardo Petroni
- Austin C. King
- Melissa S. Kim
- Rina Matsuda
- Olivia Hanselman
- Alice E. Shin
- María F. Carrera Rodríguez
- Igor E. Brodsky
- Anil Rustgi
- Ning Li
- Christopher J. Lengner
- M. Andres Blanco
2025-11-11
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.