期刊
GENES & DEVELOPMENT
卷 30, 期 12, 页码 1470-1480出版社
COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1101/gad.279190.116
关键词
somatic gene editing; breast cancer; invasive lobular carcinoma; CRISPR/Cas9; intraductal injection; mouse models
资金
- Netherlands Organization for Scientific Research (NWO)
- Netherlands Organization for Scientific Research (Cancer Genomics Netherlands [CGCNL])
- Netherlands Organization for Scientific Research (Cancer Systems Biology Center [CSBC] VENI) [016156012]
- Netherlands Organization for Scientific Research (Netherlands Genomics Initiative Zenith) [93512009, VICI 91814643]
- Worldwide Cancer Research [14-0288]
- European Union [260791, 312325]
- European Research Council (ERC Synergy project Combat-Cancer)
- National Roadmap grant for Large-Scale Research Facilities from the NWO
- Worldwide Cancer Research [14-0288] Funding Source: researchfish
Large-scale sequencing studies are rapidly identifying putative oncogenic mutations in human tumors. However, discrimination between passenger and driver events in tumorigenesis remains challenging and requires in vivo validation studies in reliable animal models of human cancer. In this study, we describe a novel strategy for in vivo validation of candidate tumor suppressors implicated in invasive lobular breast carcinoma (ILC), which is hallmarked by loss of the cell-cell adhesion molecule E-cadherin. We describe an approach to model ILC by intraductal injection of lentiviral vectors encoding Cre recombinase, the CRISPR/Cas9 system, or both in female mice carrying conditional alleles of the Cdh1 gene, encoding for E-cadherin. Using this approach, we were able to target ILC-initiating cells and induce specific gene disruption of Pten by CRISPR/Cas9-mediated somatic gene editing. Whereas intraductal injection of Cas9-encoding lentiviruses induced Cas9-specific immune responses and development of tumors that did not resemble ILC, lentiviral delivery of a Pten targeting single-guide RNA (sgRNA) in mice with mammary gland-specific loss of E-cadherin and expression of Cas9 efficiently induced ILC development. This versatile platform can be used for rapid in vivo testing of putative tumor suppressor genes implicated in ILC, providing new opportunities for modeling invasive lobular breast carcinoma in mice.
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