title: Modeling invasive lobular breast carcinoma by CRISPR/Cas9-mediated somatic genome editing of the mammary gland.
abstract: 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.
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Stefano
authors.initials: S
authors.lastname: Annunziato
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Sjors M
authors.initials: SM
authors.lastname: Kas
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Micha
authors.initials: M
authors.lastname: Nethe
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Hatice
authors.initials: H
authors.lastname: Yücel
authors.affiliation: Transgenic Core Facility, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Jessica
authors.initials: J
authors.lastname: Del Bravo
authors.affiliation: Transgenic Core Facility, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Colin
authors.initials: C
authors.lastname: Pritchard
authors.affiliation: Transgenic Core Facility, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Rahmen
authors.initials: R
authors.lastname: Bin Ali
authors.affiliation: Preclinical Intervention Unit, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Bas
authors.initials: B
authors.lastname: van Gerwen
authors.affiliation: Preclinical Intervention Unit, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Bjørn
authors.initials: B
authors.lastname: Siteur
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Anne Paulien
authors.initials: AP
authors.lastname: Drenth
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Eva
authors.initials: E
authors.lastname: Schut
authors.affiliation: Preclinical Intervention Unit, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Marieke
authors.initials: M
authors.lastname: van de Ven
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Mirjam C
authors.initials: MC
authors.lastname: Boelens
authors.affiliation: Experimental Animal Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Sjoerd
authors.initials: S
authors.lastname: Klarenbeek
authors.affiliation: Transgenic Core Facility, Mouse Clinic for Cancer and Aging (MCCA), The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Ivo J
authors.initials: IJ
authors.lastname: Huijbers
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands;
authors.forename: Martine H
authors.initials: MH
authors.lastname: van Miltenburg
authors.affiliation: Division of Molecular Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands; Cancer Genomics Netherlands, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands.
authors.forename: Jos
authors.initials: J
authors.lastname: Jonkers
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics* / Q000503:physiopathology*; D064113:CRISPR-Cas Systems; D015820:Cadherins / Q000235:genetics; D018275:Carcinoma, Lobular / Q000235:genetics* / Q000503:physiopathology*; D004195:Disease Models, Animal; D005260:Female; D000072669:Gene Editing; D020868:Gene Silencing; D016147:Genes, Tumor Suppressor; D006801:Humans; D042361:Mammary Glands, Human / Q000503:physiopathology*; D051379:Mice
chemical_list: D015820:Cadherins
keywords: CRISPR/Cas9; breast cancer; intraductal injection; invasive lobular carcinoma; mouse models; somatic gene editing