title: Renal Cell Carcinoma Programmed Death-ligand 1, a New Direct Target of Hypoxia-inducible Factor-2 Alpha, is Regulated by von Hippel-Lindau Gene Mutation Status.
abstract: Clear cell renal cell carcinomas (ccRCC) frequently display a loss of function of the von Hippel-Lindau (VHL) gene.
  
  
  
  To elucidate the putative relationship between VHL mutation status and immune checkpoint ligand programmed death-ligand 1 (PD-L1) expression.
  
  
  
  A series of 32 renal tumors composed of 11 VHL tumor-associated and 21 sporadic RCCs were used to evaluate PD-L1 expression levels after sequencing of the three exons and exon-intron junctions of the VHL gene. The 786-O, A498, and RCC4 cell lines were used to investigate the mechanisms of PD-L1 regulation.
  
  
  
  Fisher's exact test was used for VHL mutation and Kruskal-Wallis test for PD-L1 expression. If no covariate accounted for the association of VHL and PD-L1, then a Kruskal-Wallis test was used; otherwise Cochran-Mantel-Haenzsel test was used. We also used the Fligner-Policello test to compare two medians when the distributions had different dispersions.
  
  
  
  We demonstrated that tumors from ccRCC patients with VHL biallelic inactivation (ie, loss of function) display a significant increase in PD-L1 expression compared with ccRCC tumors carrying one VHL wild-type allele. Using the inducible VHL 786-O-derived cell lines with varying hypoxia-inducible factor-2 alpha (HIF-2α) stabilization levels, we showed that PD-L1 expression levels positively correlate with VHL mutation and HIF-2α expression. Targeting HIF-2α decreased PD-L1, while HIF-2α overexpression increased PD-L1 mRNA and protein levels in ccRCC cells. Interestingly, chromatin immunoprecipitation and luciferase assays revealed a direct binding of HIF-2α to a transcriptionally active hypoxia-response element in the human PD-L1 proximal promoter in 786-O cells.
  
  
  
  Our work provides the first evidence that VHL mutations positively correlate with PD-L1 expression in ccRCC and may influence the response to ccRCC anti-PD-L1/PD-1 immunotherapy.
  
  
  
  We investigated the relationship between von Hippel-Lindau mutations and programmed death-ligand 1 expression. We demonstrated that von Hippel-Lindau mutation status significantly correlated with programmed death-ligand 1 expression in clear cell renal cell carcinomas.
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France.
authors.forename: Yosra
authors.initials: Y
authors.lastname: Messai
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France; Laboratoire de Génétique Oncologique de l'Ecole Pratique des Hautes Etudes, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Gad
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France.
authors.forename: Muhammad Zaeem
authors.initials: MZ
authors.lastname: Noman
authors.affiliation: Department of Biostatistics and Epidemiology, Gustave Roussy, Villejuif, France; Institut National de la Santé et de la Recherche Médicale, CESP, Université Paris-Sud, Villejuif, France.
authors.forename: Gwenael
authors.initials: G
authors.lastname: Le Teuff
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France; Laboratoire de Génétique Oncologique de l'Ecole Pratique des Hautes Etudes, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Couve
authors.affiliation: Laboratory of Experimental Cancer Research, Department of Oncology, Luxembourg Institute of Health, Luxembourg City, Luxembourg.
authors.forename: Bassam
authors.initials: B
authors.lastname: Janji
authors.affiliation: Department of Pathology, University Hospital of Rennes, Rennes, France; The French National Centre for Scientific Research, IGDR Biosit, Rennes 1 University, Rennes, France.
authors.forename: Solenne Florence
authors.initials: SF
authors.lastname: Kammerer
authors.affiliation: Department of Pathology, University Hospital of Rennes, Rennes, France; The French National Centre for Scientific Research, IGDR Biosit, Rennes 1 University, Rennes, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Rioux-Leclerc
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France.
authors.forename: Meriem
authors.initials: M
authors.lastname: Hasmim
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France; Université Paris-Sud, Assistance Publique-Hôpitaux de Paris, Service d'Anatomo-Pathologie, Hôpital Bicêtre, Le Kremlin-Bicêtre, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Ferlicot
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, Institut Cochin, Paris, France; The French National Center for Scientific Research, Paris, France; Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Baud
authors.affiliation: Service d'Urologie, AP-HP Hôpital Européen Georges Pompidou, Paris, France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Mejean
authors.affiliation: The Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
authors.forename: David Robert
authors.initials: DR
authors.lastname: Mole
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France; Laboratoire de Génétique Oncologique de l'Ecole Pratique des Hautes Etudes, Paris, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Richard
authors.affiliation: Cancer Institute, Gustave Roussy Cancer Campus, Grand Paris, Villejuif, France.
authors.forename: Alexander M M
authors.initials: AM
authors.lastname: Eggermont
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France; Medical Oncology Department, Gustave Roussy Cancer Campus, Villejuif, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Albiges
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France.
authors.forename: Fathia
authors.initials: F
authors.lastname: Mami-Chouaib
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France; Medical Oncology Department, Gustave Roussy Cancer Campus, Villejuif, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Escudier
authors.affiliation: INSERM (Institut National de la Santé et de la Recherche Médicale) UMR1186, Laboratory Integrative Tumor Immunology and Genetic Oncology, Villejuif, France; INSERM, Gustave Roussy, Univ. Paris-Sud, Université Paris-Saclay, Villejuif, F-94805, France. Electronic address: salem.chouaib@gustaveroussy.fr.
authors.forename: Salem
authors.initials: S
authors.lastname: Chouaib
mesh_terms: D000483:Alleles; D060890:B7-H1 Antigen / Q000235:genetics* / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics*; D002292:Carcinoma, Renal Cell / Q000235:genetics*; D045744:Cell Line, Tumor; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics*; D000073658:Loss of Function Mutation; D019656:Loss of Heterozygosity; D008297:Male; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics*; D000099162:Endothelial PAS Domain-Containing Protein 1
chemical_list: D060890:B7-H1 Antigen; D051792:Basic Helix-Loop-Helix Proteins; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D000099162:Endothelial PAS Domain-Containing Protein 1; C423236:CD274 protein, human
keywords: HIF-2α; PD-L1; VHL mutations; ccRCC