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

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.

European urology. 2015 Dec 17 [Epub ahead of print]

Yosra Messai, Sophie Gad, Muhammad Zaeem Noman, Gwenael Le Teuff, Sophie Couve, Bassam Janji, Solenne Florence Kammerer, Nathalie Rioux-Leclerc, Meriem Hasmim, Sophie Ferlicot, Véronique Baud, Arnaud Mejean, David Robert Mole, Stéphane Richard, Alexander M M Eggermont, Laurence Albiges, Fathia Mami-Chouaib, Bernard Escudier, Salem Chouaib

PubMed

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. , 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. , 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. , 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. , 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. , Laboratory of Experimental Cancer Research, Department of Oncology, Luxembourg Institute of Health, Luxembourg City, Luxembourg. , Department of Pathology, University Hospital of Rennes, Rennes, France; The French National Centre for Scientific Research, IGDR Biosit, Rennes 1 University, Rennes, France. , Department of Pathology, University Hospital of Rennes, Rennes, France; The French National Centre for Scientific Research, IGDR Biosit, Rennes 1 University, Rennes, France. , 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. , 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. , 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. , Service d'Urologie, AP-HP Hôpital Européen Georges Pompidou, Paris, France. , The Henry Wellcome Building for Molecular Physiology, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom. , 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. , Cancer Institute, Gustave Roussy Cancer Campus, Grand Paris, Villejuif, France. , 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. , 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. , 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. , 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.