CD271 Expression on Patient Melanoma Cells Is Unstable and Unlinked to Tumorigenicity.
The stability of markers that identify cancer cells that propagate disease is important to the outcomes of targeted therapy strategies. In human melanoma, conflicting data exist as to whether hierarchical expression of CD271/p75/NGFR (nerve growth factor receptor) marks cells with enriched tumorigenicity, which would compel their specific targeting in therapy. To test whether these discrepancies relate to differences among groups in assay approaches, we undertook side-by-side testing of published methods of patient-derived melanoma xenografting (PDX), including comparisons of tissue digestion procedures or coinjected Matrigel formulations. We found that CD271(-) and CD271(+) melanoma cells from each of seven patients were similarly tumorigenic, regardless of assay variations. Surprisingly variable CD271 expression patterns were observed in the analyses of sibling PDX tumors (n = 68) grown in the same experiments from either CD271(-) or CD271(+) cells obtained from patients. This indicates unstable intratumoral lineage relationships between CD271(-) and CD271(+) melanoma cells that are inconsistent with classical, epigenetically based theories of disease progression, such as the cancer stem cell and plasticity models. SNP genotyping of pairs of sibling PDX tumors grown from phenotypically identical CD271(-) or CD271(+) cells showed large pairwise differences in copy number (28%-48%). Differences were also apparent in the copy number profiles of CD271(-) and CD271(+) cells purified directly from each of the four melanomas (1.4%-23%). Thus, CD271 expression in patient melanomas is unstable, not consistently linked to increased tumorigenicity and associated with genetic heterogeneity, undermining its use as a marker in clinical studies. Cancer Res; 76(13); 3965-77. ©2016 AACR.
Cancer Development and Treatment Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia. Sir Peter MacCallum Department of Pathology, University of Melbourne, Parkville, Australia.
Samantha E
SE
Boyle
Cancer Development and Treatment Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia. Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, Australia.
Clare G
CG
Fedele
Bioinformatics and Cancer Genomics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia. Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.
Vincent
V
Corbin
Cancer Development and Treatment Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia. Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, Australia.
Elisha
E
Wybacz
Cancer Development and Treatment Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia. Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, Australia.
Pacman
P
Szeto
Department of Cancer Medicine, Peter MacCallum Cancer Centre, East Melbourne, Australia.
Jeremy
J
Lewin
Translational Research Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia.
Richard J
RJ
Young
Department of Cancer Medicine, Peter MacCallum Cancer Centre, East Melbourne, Australia.
Annie
A
Wong
Bioinformatics and Cancer Genomics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia. Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.
Robert
R
Fuller
Department of Surgery, Peter MacCallum Cancer Centre, East Melbourne, Australia.
John
J
Spillane
Department of Surgery, Peter MacCallum Cancer Centre, East Melbourne, Australia.
David
D
Speakman
Department of Surgery, Peter MacCallum Cancer Centre, East Melbourne, Australia.
Simon
S
Donahoe
Department of Surgery, Peter MacCallum Cancer Centre, East Melbourne, Australia.
Miklos
M
Pohl
Department of Surgery, Peter MacCallum Cancer Centre, East Melbourne, Australia.
David
D
Gyorki
Department of Surgery, Peter MacCallum Cancer Centre, East Melbourne, Australia.
Michael A
MA
Henderson
Sir Peter MacCallum Department of Pathology, University of Melbourne, Parkville, Australia. Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, Australia. Gene Regulation Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia.
Ricky W
RW
Johnstone
Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, Australia. Bioinformatics and Cancer Genomics Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia. Bioinformatics Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.
Anthony T
AT
Papenfuss
Cancer Development and Treatment Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Australia. Sir Peter MacCallum Department of Pathology, University of Melbourne, Parkville, Australia. Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, Australia. Department of Cancer Medicine, Peter MacCallum Cancer Centre, East Melbourne, Australia. mark.shackleton@petermac.org.
Mark
M
Shackleton
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D008545:Melanoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D017475:Receptors, Nerve Growth Factor / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
C484065:NGFR protein, human; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D017475:Receptors, Nerve Growth Factor