In vivo genome-editing screen identifies tumor suppressor genes that cooperate with Trp53 loss during mammary tumorigenesis.

Autor: Heitink L; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Medical Biology, The University of Melbourne, Parkville, Australia., Whittle JR; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Medical Biology, The University of Melbourne, Parkville, Australia.; Department of Medical Oncology, Peter MacCallum Cancer Centre, Melbourne, Australia., Vaillant F; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Medical Biology, The University of Melbourne, Parkville, Australia., Capaldo BD; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Medical Biology, The University of Melbourne, Parkville, Australia., Dekkers JF; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Princess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands., Dawson CA; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Medical Biology, The University of Melbourne, Parkville, Australia.; Immunology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia., Milevskiy MJG; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Medical Biology, The University of Melbourne, Parkville, Australia., Surgenor E; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia., Tsai M; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia., Chen HR; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia., Christie M; Personalised Oncology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Pathology, The Royal Melbourne Hospital, Parkville, Australia., Chen Y; Department of Medical Biology, The University of Melbourne, Parkville, Australia.; Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia., Smyth GK; Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; School of Mathematics and Statistics, The University of Melbourne, Parkville, Australia., Herold MJ; Department of Medical Biology, The University of Melbourne, Parkville, Australia.; Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia., Strasser A; Department of Medical Biology, The University of Melbourne, Parkville, Australia.; Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia., Lindeman GJ; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Medical Biology, The University of Melbourne, Parkville, Australia.; Department of Medical Oncology, Peter MacCallum Cancer Centre, Melbourne, Australia., Visvader JE; ACRF Cancer Biology and Stem Cells Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.; Department of Medical Biology, The University of Melbourne, Parkville, Australia.
Jazyk: angličtina
Zdroj: Molecular oncology [Mol Oncol] 2022 Mar; Vol. 16 (5), pp. 1119-1131. Date of Electronic Publication: 2022 Jan 26.
DOI: 10.1002/1878-0261.13179
Abstrakt: Breast cancer is a heterogeneous disease that comprises multiple histological and molecular subtypes. To gain insight into mutations that drive breast tumorigenesis, we describe a pipeline for the identification and validation of tumor suppressor genes. Based on an in vivo genome-wide CRISPR/Cas9 screen in Trp53 +/- heterozygous mice, we identified tumor suppressor genes that included the scaffold protein Axin1, the protein kinase A regulatory subunit gene Prkar1a, as well as the proof-of-concept genes Pten, Nf1, and Trp53 itself. Ex vivo editing of primary mammary epithelial organoids was performed to further interrogate the roles of Axin1 and Prkar1a. Increased proliferation and profound changes in mammary organoid morphology were observed for Axin1/Trp53 and Prkar1a/Trp53 double mutants compared to Pten/Trp53 double mutants. Furthermore, direct in vivo genome editing via intraductal injection of lentiviruses engineered to express dual short-guide RNAs revealed that mutagenesis of Trp53 and either Prkar1a, Axin1, or Pten markedly accelerated tumor development compared to Trp53-only mutants. This proof-of-principle study highlights the application of in vivo CRISPR/Cas9 editing for uncovering cooperativity between defects in tumor suppressor genes that elicit mammary tumorigenesis.
(© 2022 The Authors. Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)
Databáze: MEDLINE
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