In vivo genome editing and organoid transplantation models of colorectal cancer and metastasis
Cette étude présente une méthode utilisant la technologie CRISPR-Cas9 d'ingénierie du génome pour analyser rapidement, à l'aide de greffes d'organoïdes sur des modèles murins, les mécanismes par lesquels des gènes sont impliqués dans la progression d'une tumeur colorectale ou dans le processus métastatique
In vivo interrogation of the function of genes implicated in tumorigenesis is limited by the need to generate and cross germline mutant mice. Here we describe approaches to model colorectal cancer (CRC) and metastasis, which rely on in situ gene editing and orthotopic organoid transplantation in mice without cancer-predisposing mutations. Autochthonous tumor formation is induced by CRISPR-Cas9-based editing of the Apc and Trp53 tumor suppressor genes in colon epithelial cells and by orthotopic transplantation of Apc-edited colon organoids. Apc
Δ/Δ;KrasG12D/+;Trp53Δ/Δ (AKP) mouse colon organoids and human CRC organoids engraft in the distal colon and metastasize to the liver. Finally, we apply the orthotopic transplantation model to characterize the clonal dynamics of Lgr5+ stem cells and demonstrate sequential activation of an oncogene in established colon adenomas. These experimental systems enable rapid in vivo characterization of cancer-associated genes and reproduce the entire spectrum of tumor progression and metastasis.
Nature Biotechnology , résumé, 2016