title: Pancreatic cancer modeling using retrograde viral vector delivery and in vivo CRISPR/Cas9-mediated somatic genome editing.
abstract: Pancreatic ductal adenocarcinoma (PDAC) is a genomically diverse, prevalent, and almost invariably fatal malignancy. Although conventional genetically engineered mouse models of human PDAC have been instrumental in understanding pancreatic cancer development, these models are much too labor-intensive, expensive, and slow to perform the extensive molecular analyses needed to adequately understand this disease. Here we demonstrate that retrograde pancreatic ductal injection of either adenoviral-Cre or lentiviral-Cre vectors allows titratable initiation of pancreatic neoplasias that progress into invasive and metastatic PDAC. To enable in vivo CRISPR/Cas9-mediated gene inactivation in the pancreas, we generated a Cre-regulated Cas9 allele and lentiviral vectors that express Cre and a single-guide RNA. CRISPR-mediated targeting of Lkb1 in combination with oncogenic Kras expression led to selection for inactivating genomic alterations, absence of Lkb1 protein, and rapid tumor growth that phenocopied Cre-mediated genetic deletion of Lkb1. This method will transform our ability to rapidly interrogate gene function during the development of this recalcitrant cancer.
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Shin-Heng
authors.initials: SH
authors.lastname: Chiou
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Ian P
authors.initials: IP
authors.lastname: Winters
authors.affiliation: Department of Developmental Biology, Stanford University School of Medicine, Stanford, California 94305, USA; Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Santiago
authors.initials: S
authors.lastname: Naranjo
authors.affiliation: Rutgers Cancer Institute of New Jersey, New Brunswick, New Jersey 08903, USA; Department of Surgery, Rutgers Robert Wood Johnson University Medical School, New Brunswick, New Jersey 08903, USA; Department of Pharmacology, Rutgers Robert Wood Johnson University Medical School, New Brunswick, New Jersey 08903, USA;
authors.forename: Crissy
authors.initials: C
authors.lastname: Dudgeon
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Fiona B
authors.initials: FB
authors.lastname: Tamburini
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Brady
authors.affiliation: Cancer Biology Program, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Dian
authors.initials: D
authors.lastname: Yang
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Barbara M
authors.initials: BM
authors.lastname: Grüner
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Chen-Hua
authors.initials: CH
authors.lastname: Chuang
authors.affiliation: Cancer Biology Program, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Deborah R
authors.initials: DR
authors.lastname: Caswell
authors.affiliation: Stanford Cancer Institute, Stanford University School of Medicine, Stanford, California 94305, USA; Transgenic, Knockout, and Tumor Model Center, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Hong
authors.initials: H
authors.lastname: Zeng
authors.affiliation: Department of Comparative Medicine, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Pauline
authors.initials: P
authors.lastname: Chu
authors.affiliation: Department of Pathology, University of California at San Francisco, San Francisco, California 94143, USA;
authors.forename: Grace E
authors.initials: GE
authors.lastname: Kim
authors.affiliation: Rutgers Cancer Institute of New Jersey, New Brunswick, New Jersey 08903, USA; Department of Surgery, Rutgers Robert Wood Johnson University Medical School, New Brunswick, New Jersey 08903, USA; Department of Pharmacology, Rutgers Robert Wood Johnson University Medical School, New Brunswick, New Jersey 08903, USA;
authors.forename: Darren R
authors.initials: DR
authors.lastname: Carpizo
authors.affiliation: Department of Developmental Biology, Stanford University School of Medicine, Stanford, California 94305, USA; Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California 94305, USA;
authors.forename: Seung K
authors.initials: SK
authors.lastname: Kim
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA; Cancer Biology Program, Stanford University School of Medicine, Stanford, California 94305, USA; Stanford Cancer Institute, Stanford University School of Medicine, Stanford, California 94305, USA; Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, USA.
authors.forename: Monte M
authors.initials: MM
authors.lastname: Winslow
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000503:physiopathology*; D000818:Animals; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics / Q000503:physiopathology*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004195:Disease Models, Animal; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000235:genetics; D016678:Genome / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic
keywords: CRISPR; genome editing; mouse model; pancreatic cancer