A CRISPR/Cas9-Engineered ARID1A -Deficient Human Gastric Cancer Organoid Model Reveals Essential and Nonessential Modes of Oncogenic Transformation

• Published in: Cancer discovery Vol. 11; no. 6; p. 1562
• Main Authors: Lo, Yuan-Hung, Kolahi, Kevin S, Du, Yuhong, Chang, Chiung-Ying, Krokhotin, Andrey, Nair, Ajay, Sobba, Walter D, Karlsson, Kasper, Jones, Sunny J, Longacre, Teri A, Mah, Amanda T, Tercan, Bahar, Sockell, Alexandra, Xu, Hang, Seoane, Jose A, Chen, Jin, Shmulevich, Ilya, Weissman, Jonathan S, Curtis, Christina, Califano, Andrea, Fu, Haian, Crabtree, Gerald R, Kuo, Calvin J
• Format: Journal Article
• Language: English
• Published: United States 01-06-2021
• Subjects: Cell Transformation, Neoplastic; CRISPR-Cas Systems; DNA-Binding Proteins - genetics; Humans; Models, Biological; Mutation; Stomach Neoplasms - genetics; Transcription Factors - genetics
• ISSN: 2159-8290
• DOI: 10.1158/2159-8290.cd-20-1109

Mutations in
rank among the most common molecular aberrations in human cancer. However, oncogenic consequences of
mutation in human cells remain poorly defined due to lack of forward genetic models. Here, CRISPR/Cas9-mediated
knockout (KO) in primary
human gastric organoids induced morphologic dysplasia, tumorigenicity, and mucinous differentiation. Genetic WNT/β-catenin activation rescued mucinous differentiation, but not hyperproliferation, suggesting alternative pathways of
KO-mediated transformation.
mutation induced transcriptional regulatory modules characteristic of microsatellite instability and Epstein-Barr virus-associated subtype human gastric cancer, including
-associated mitotic genes and
survivin. Convergently, high-throughput compound screening indicated selective vulnerability of
-deficient organoids to inhibition of BIRC5/survivin, functionally implicating this pathway as an essential mediator of
KO-dependent early-stage gastric tumorigenesis. Overall, we define distinct pathways downstream of oncogenic
mutation, with nonessential WNT-inhibited mucinous differentiation in parallel with essential transcriptional
-stimulated proliferation, illustrating the general utility of organoid-based forward genetic cancer analysis in human cells. SIGNIFICANCE: We establish the first human forward genetic modeling of a commonly mutated tumor suppressor gene,
. Our study integrates diverse modalities including CRISPR/Cas9 genome editing, organoid culture, systems biology, and small-molecule screening to derive novel insights into early transformation mechanisms of
-deficient gastric cancers.
.
.