Isolation and characterization of renal cancer stem cells from patient-derived xenografts
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Meriem Hasmim1,2,*, Stefania Bruno3,*, Sandy Azzi2, Cindy Gallerne2, Julien Giron Michel2, Giulia Chiabotto4, Vincent Lecoz2, Cristina Romei5, Grazia Maria Spaggiari5, Annalisa Pezzolo6, Vito Pistoia6, Eric Angevin1,7, Sophie Gad1,8, Sophie Ferlicot1,9, Yosra Messai1, Claudine Kieda10, Denis Clay11, Federica Sabatini12, Bernard Escudier1,7, Giovanni Camussi4, Pierre Eid2, Bruno Azzarone5, Salem Chouaib1
1INSERM U 1186, Equipe labellisée Ligue Contre le Cancer, Gustave Roussy Campus, Villejuif, France
2INSERM UMR 1014, Lavoisier Building, Paul Brousse Hospital, Villejuif, France
3Department of Molecular Biotechnology and Healthy Science, Molecular Biotechnology Center, University of Torino, Turin, Italy
4Department of Medical Science, University of Torino, Medical School, Torino, Italy
5DIMES, UNIGE, Genova, Italy
6Laboratory of Oncology Giannina Gaslini Institute, Genoa, Italy
7Medical Oncology Department, Gustave Roussy Campus, Villejuif, France
8Laboratoire de Génétique Oncologique EPHE, Ecole Pratique des Hautes Etudes, Paris, France
9Université Paris-Sud, Assistance Publique-Hôpitaux de Paris, Service d'Anatomo-Pathologie, Hôpital Bicêtre, Le Kremlin-Bicêtre, France
10Centre de Biophysique Moléculaire, CNRS UPR 4301, Orléans, France
11INSERM UMR 972, Paul Brousse Hospital, Villejuif, France
12Stem Cell and Cell Therapy Laboratory, Istituto G. Gaslini, Genoa, Italy
*These authors have contributed equally to this work
Salem Chouaib, e-mail: email@example.com
Bruno Azzarone, e-mail: firstname.lastname@example.org
Keywords: clear cell renal cell carcinoma, cancer stem cells, patient-derived xenografts, CD133, EpCAM
Received: July 24, 2015 Accepted: October 08, 2015 Published: November 02, 2015
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.
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