title: Isolation and characterization of renal cancer stem cells from patient-derived xenografts.
abstract: As rapidly developing patient-derived xenografts (PDX) could represent potential sources of cancer stem cells (CSC), we selected and characterized non-cultured PDX cell suspensions from four different renal carcinomas (RCC). Only the cell suspensions from the serial xenografts (PDX-1 and PDX-2) of an undifferentiated RCC (RCC-41) adapted to the selective CSC medium. The cell suspension derived from the original tumor specimen (RCC-41-P-0) did not adapt to the selective medium and strongly expressed CSC-like markers (CD133 and CD105) together with the non-CSC tumor marker E-cadherin. In comparison, PDX-1 and PDX-2 cells exhibited evolution in their phenotype since PDX-1 cells were CD133high/CD105-/Ecadlow and PDX-2 cells were CD133low/CD105-/Ecad-. Both PDX subsets expressed additional stem cell markers (CD146/CD29/OCT4/NANOG/Nestin) but still contained non-CSC tumor cells. Therefore, using different cell sorting strategies, we characterized 3 different putative CSC subsets (RCC-41-PDX-1/CD132+, RCC-41-PDX-2/CD133-/EpCAMlow and RCC-41-PDX-2/CD133+/EpCAMbright). In addition, transcriptomic analysis showed that RCC-41-PDX-2/CD133- over-expressed the pluripotency gene ERBB4, while RCC-41-PDX-2/CD133+ over-expressed several tumor suppressor genes. These three CSC subsets displayed ALDH activity, formed serial spheroids and developed serial tumors in SCID mice, although RCC-41-PDX-1/CD132+ and RCC-41-PDX-2/CD133+ displayed less efficiently the above CSC properties. RCC-41-PDX-1/CD132+ tumors showed vessels of human origin with CSC displaying peri-vascular distribution. By contrast, RCC-41-PDX-2 originated tumors exhibiting only vessels of mouse origin without CSC peri-vascular distribution.Altogether, our results indicate that PDX murine microenvironment promotes a continuous redesign of CSC phenotype, unmasking CSC subsets potentially present in a single RCC or generating ex novo different CSC-like subsets.
authors.affiliation: INSERM U 1186, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy Campus, Villejuif, France.|INSERM UMR 1014, Lavoisier Building, Paul Brousse Hospital, Villejuif, France.
authors.forename: Meriem
authors.initials: M
authors.lastname: Hasmim
authors.affiliation: Department of Molecular Biotechnology and Healthy Science, Molecular Biotechnology Center, University of Torino, Turin, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Bruno
authors.affiliation: INSERM UMR 1014, Lavoisier Building, Paul Brousse Hospital, Villejuif, France.
authors.forename: Sandy
authors.initials: S
authors.lastname: Azzi
authors.affiliation: INSERM UMR 1014, Lavoisier Building, Paul Brousse Hospital, Villejuif, France.
authors.forename: Cindy
authors.initials: C
authors.lastname: Gallerne
authors.affiliation: INSERM UMR 1014, Lavoisier Building, Paul Brousse Hospital, Villejuif, France.
authors.forename: Julien Giron
authors.initials: JG
authors.lastname: Michel
authors.affiliation: Department of Medical Science, University of Torino, Medical School, Torino, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Chiabotto
authors.affiliation: INSERM UMR 1014, Lavoisier Building, Paul Brousse Hospital, Villejuif, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Lecoz
authors.affiliation: DIMES, UNIGE, Genova, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Romei
authors.affiliation: DIMES, UNIGE, Genova, Italy.
authors.forename: Grazia Maria
authors.initials: GM
authors.lastname: Spaggiari
authors.affiliation: Laboratory of Oncology Giannina Gaslini Institute, Genoa, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Pezzolo
authors.affiliation: Laboratory of Oncology Giannina Gaslini Institute, Genoa, Italy.
authors.forename: Vito
authors.initials: V
authors.lastname: Pistoia
authors.affiliation: INSERM U 1186, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy Campus, Villejuif, France.|Medical Oncology Department, Gustave Roussy Campus, Villejuif, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Angevin
authors.affiliation: INSERM U 1186, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy Campus, Villejuif, France.|Laboratoire de Génétique Oncologique EPHE, Ecole Pratique des Hautes Etudes, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Gad
authors.affiliation: INSERM U 1186, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy Campus, Villejuif, 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: INSERM U 1186, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy Campus, Villejuif, France.
authors.forename: Yosra
authors.initials: Y
authors.lastname: Messai
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR 4301, Orléans, France.
authors.forename: Claudine
authors.initials: C
authors.lastname: Kieda
authors.affiliation: INSERM UMR 972, Paul Brousse Hospital, Villejuif, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Clay
authors.affiliation: Stem Cell and Cell Therapy Laboratory, Istituto G. Gaslini, Genoa, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Sabatini
authors.affiliation: INSERM U 1186, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy Campus, Villejuif, France.|Medical Oncology Department, Gustave Roussy Campus, Villejuif, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Escudier
authors.affiliation: Department of Medical Science, University of Torino, Medical School, Torino, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Camussi
authors.affiliation: INSERM UMR 1014, Lavoisier Building, Paul Brousse Hospital, Villejuif, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Eid
authors.affiliation: DIMES, UNIGE, Genova, Italy.
authors.forename: Bruno
authors.initials: B
authors.lastname: Azzarone
authors.affiliation: INSERM U 1186, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy Campus, Villejuif, France.
authors.forename: Salem
authors.initials: S
authors.lastname: Chouaib
mesh_terms: D000818:Animals; D002292:Carcinoma, Renal Cell / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D064593:Heterografts; D006801:Humans; D007680:Kidney Neoplasms / Q000473:pathology*; D051379:Mice; D016513:Mice, SCID; D014411:Neoplastic Stem Cells / Q000473:pathology*; D014407:Tumor Cells, Cultured
keywords: CD133; EpCAM; cancer stem cells; clear cell renal cell carcinoma; patient-derived xenografts