Isolation and characterization of renal cancer stem cells from patient-derived xenografts

Meriem Hasmim, Stefania Bruno, Sandy Azzi, Cindy Gallerne, Julien Giron Michel, Giulia Chiabotto, Vincent Lecoz, Cristina Romei, Grazia Maria Spaggiari, Annalisa Pezzolo, Vito Pistoia, Eric Angevin, Sophie Gad, Sophie Ferlicot, Yosra Messai, Claudine Kieda, Denis Clay, Federica Sabatini, Bernard Escudier, Giovanni CamussiPierre Eid, Bruno Azzarone, Salem Chouaib

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)15507-15524
Number of pages18
JournalOncotarget
Volume7
Issue number13
DOIs
Publication statusPublished - Mar 29 2016

Keywords

  • Cancer stem cells
  • CD133
  • Clear cell renal cell carcinoma
  • EpCAM
  • Patient-derived xenografts

ASJC Scopus subject areas

  • Oncology

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  • Cite this

    Hasmim, M., Bruno, S., Azzi, S., Gallerne, C., Michel, J. G., Chiabotto, G., Lecoz, V., Romei, C., Spaggiari, G. M., Pezzolo, A., Pistoia, V., Angevin, E., Gad, S., Ferlicot, S., Messai, Y., Kieda, C., Clay, D., Sabatini, F., Escudier, B., ... Chouaib, S. (2016). Isolation and characterization of renal cancer stem cells from patient-derived xenografts. Oncotarget, 7(13), 15507-15524. https://doi.org/10.18632/oncotarget.6266