title: CRISPR-Mediated VHL Knockout Generates an Improved Model for Metastatic Renal Cell Carcinoma.
abstract: Metastatic renal cell carcinoma (mRCC) is nearly incurable and accounts for most of the mortality associated with RCC. Von Hippel Lindau (VHL) is a tumour suppressor that is lost in the majority of clear cell RCC (ccRCC) cases. Its role in regulating hypoxia-inducible factors-1α (HIF-1α) and -2α (HIF-2α) is well-studied. Recent work has demonstrated that VHL knock down induces an epithelial-mesenchymal transition (EMT) phenotype. In this study we showed that a CRISPR/Cas9-mediated knock out of VHL in the RENCA model leads to morphologic and molecular changes indicative of EMT, which in turn drives increased metastasis to the lungs. RENCA cells deficient in HIF-1α failed to undergo EMT changes upon VHL knockout. RNA-seq revealed several HIF-1α-regulated genes that are upregulated in our VHL knockout cells and whose overexpression signifies an aggressive form of ccRCC in the cancer genome atlas (TCGA) database. Independent validation in a new clinical dataset confirms the upregulation of these genes in ccRCC samples compared to adjacent normal tissue. Our findings indicate that loss of VHL could be driving tumour cell dissemination through stabilization of HIF-1α in RCC. A better understanding of the mechanisms involved in this phenomenon can guide the search for more effective treatments to combat mRCC.
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles CA 90095, USA.
authors.forename: Shiruyeh
authors.initials: S
authors.lastname: Schokrpur
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles CA 90095, USA.|Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.|Department of Paediatric Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Junhui
authors.initials: J
authors.lastname: Hu
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles CA 90095, USA.
authors.forename: Diana L
authors.initials: DL
authors.lastname: Moughon
authors.affiliation: Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Peijun
authors.initials: P
authors.lastname: Liu
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles CA 90095, USA.
authors.forename: Lucia C
authors.initials: LC
authors.lastname: Lin
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles CA 90095, USA.
authors.forename: Kip
authors.initials: K
authors.lastname: Hermann
authors.affiliation: Department of Computer Science and Human Genetics, University of California at Los Angeles CA 90095, USA.
authors.forename: Serghei
authors.initials: S
authors.lastname: Mangul
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles CA 90095, USA.|Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Guan
authors.affiliation: Department of Computer Science and Human Genetics, University of California at Los Angeles CA 90095, USA.|Department of Molecular, Cell, and Developmental Biology, University of California at Los Angeles CA 90095, USA.
authors.forename: Matteo
authors.initials: M
authors.lastname: Pellegrini
authors.affiliation: Department of Urology and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles CA 90095, USA.|Department of Urology, David Geffen School of Medicine, University of California at Los Angeles CA 90095, USA.
authors.forename: Lily
authors.initials: L
authors.lastname: Wu
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D000076987:CRISPR-Associated Protein 9; D064113:CRISPR-Cas Systems; D002292:Carcinoma, Renal Cell / Q000235:genetics / Q000556:secondary*; D045744:Cell Line, Tumor; D002465:Cell Movement; D066264:Datasets as Topic; D004195:Disease Models, Animal; D004720:Endonucleases; D058750:Epithelial-Mesenchymal Transition; D005260:Female; D000072669:Gene Editing; D015972:Gene Expression Regulation, Neoplastic; D064593:Heterografts; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000502:physiology; D007680:Kidney Neoplasms / Q000473:pathology*; D008175:Lung Neoplasms / Q000556:secondary; D051379:Mice; D018345:Mice, Knockout; D008819:Mice, Nude; D009363:Neoplasm Proteins / Q000235:genetics / Q000502:physiology; D000094704:RNA, Guide, CRISPR-Cas Systems; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D009363:Neoplasm Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases; C497446:VHL protein, human; C494469:VHL protein, mouse