Abstract 4305: Refinement of the glioma cancer stem cell marker profile
Authors: Lindsey D. Goodman, Feng Gao, Joy Gumin, Diana Villareal, Ravesanker Ezhilarasan, Patrice Love, Howard Colman, Frederick F. Lang, Kenneth D. Aldape, Erik P. Sulman
Journal: Cancer Research (2010)

Abstract Infiltrating gliomas are the most frequently occurring adult brain tumor and, among high-grade forms, typically result in poor patient survival. These tumors tend to be refractory to most treatments, including aggressive surgical resection and chemoradiation. This treatment resistance is hypothesized to be the result of a small population of cells within the tumor which are responsible for tumor initiation, called glioma cancer stem cells (GCSCs). As is true in other stem cell systems, such as hematopoiesis, multiple cell-surface protein markers are needed to accurately define the true stem cell population. We and others have previously identified the glycoprotein podoplanin (pdpn) as a marker of GCSC, which improves upon the established marker, CD133. To refine the GCSC marker profile beyond these two proteins, we examined a number of candidate markers in combination with pdpn that have been proposed as stem cell markers in gliomas or other tumors or tissues. Candidate markers were analyzed by flow cytometry using primary glioma cell lines maintained as neurosphere cultures and fresh tumor specimens. Markers identifying a unique cell population either alone or in combination with pdpn were further analyzed by in vitro neurosphere formation assay and clonogenic survival assay following 2 Gy ionizing radiation. Candidates showing enrichment for stem cells based on these assays were examine for in vivo tumor initiation using flow sorted subpopulations of cells with and without the expression of the candidate protein. As expression of putative stem cell markers will likely predict for poor patient survival, candidate markers were also examined in human tumors by immunohistochemsitry. Among the most promosing candidates were CD97, CXCR4, CD58 and CD44. Flow cytometry revealed CD97 expression in 75%, CD58 in 80%, CXCR4 in 100%, and CD44 in 100% of tumors examined to date. Examination of primary neurosphere cultures, found CD97 in 80%, CXCR4 in 83%, CD58 in 100%, and CD44 in 100% of lines examined. Both CD44+ and CD58+ cells exhibited increased neurophere formation rates and in vivo tumor initiation. These stem-like characteristics were enhanced in those subpopulations also expressing pdpn. RNAs collected from these subpopulations were used for expression profiling to determine if these subpopulation express a unique gene expression. Results thus far indicate that pdpn expression is needed for the stem-like phenotype and that this is enhanced in populations co-expressing CD44, CD15, and/or CD58. Further refinement of this profile will aide in the isolation of the “true” GCSC and allow for improved therapeutic targeting of these resistant cells. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4305.