Limits...
Glial progenitor-like phenotype in low-grade glioma and enhanced CD133-expression and neuronal lineage differentiation potential in high-grade glioma.

Rebetz J, Tian D, Persson A, Widegren B, Salford LG, Englund E, Gisselsson D, Fan X - PLoS ONE (2008)

Bottom Line: In parallel, the expression of glial fibrillary acidic protein (GFAP), synaptophysin and neuron-specific enolase (NSE) was immunohistochemically analyzed in fixed tissue specimens.The rare CD133 expressing cells in low-grade glioma specimens typically co-expressed vessel endothelial marker CD31.The CD133 expression correlates inversely with length of patient survival.

View Article: PubMed Central - PubMed

Affiliation: The Rausing Laboratory, Division of Neurosurgery, Lund University Hospital, Lund, Sweden.

ABSTRACT

Background: While neurosphere- as well as xenograft tumor-initiating cells have been identified in gliomas, the resemblance between glioma cells and neural stem/progenitor cells as well as the prognostic value of stem/progenitor cell marker expression in glioma are poorly clarified.

Methodology/principal findings: Viable glioma cells were characterized for surface marker expression along the glial genesis hierarchy. Six low-grade and 17 high-grade glioma specimens were flow-cytometrically analyzed for markers characteristics of stem cells (CD133); glial progenitors (PDGFRalpha, A2B5, O4, and CD44); and late oligodendrocyte progenitors (O1). In parallel, the expression of glial fibrillary acidic protein (GFAP), synaptophysin and neuron-specific enolase (NSE) was immunohistochemically analyzed in fixed tissue specimens. Irrespective of the grade and morphological diagnosis of gliomas, glioma cells concomitantly expressed PDGFRalpha, A2B5, O4, CD44 and GFAP. In contrast, O1 was weakly expressed in all low-grade and the majority of high-grade glioma specimens analyzed. Co-expression of neuronal markers was observed in all high-grade, but not low-grade, glioma specimens analyzed. The rare CD133 expressing cells in low-grade glioma specimens typically co-expressed vessel endothelial marker CD31. In contrast, distinct CD133 expression profiles in up to 90% of CD45-negative glioma cells were observed in 12 of the 17 high-grade glioma specimens and the majority of these CD133 expressing cells were CD31 negative. The CD133 expression correlates inversely with length of patient survival. Surprisingly, cytogenetic analysis showed that gliomas contained normal and abnormal cell karyotypes with hitherto indistinguishable phenotype.

Conclusions/significance: This study constitutes an important step towards clarification of lineage commitment and differentiation blockage of glioma cells. Our data suggest that glioma cells may resemble expansion of glial lineage progenitor cells with compromised differentiation capacity downstream of A2B5 and O4 expression. The concurrent expression of neuronal markers demonstrates that high-grade glioma cells are endowed with multi-lineage differentiation potential in vivo. Importantly, enhanced CD133 expression marks a poor prognosis in gliomas.

Show MeSH

Related in: MedlinePlus

Neuronal marker expression was not detected in low-grade gliomas, but in high-grade gliomas.Staining patterns of GFAP and NSE expression from representative low-glioma (patient ID: #2) and high-grade glioma ((patient ID: #12 and #19) are shown. Original magnification: ×100
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2277459&req=5

pone-0001936-g002: Neuronal marker expression was not detected in low-grade gliomas, but in high-grade gliomas.Staining patterns of GFAP and NSE expression from representative low-glioma (patient ID: #2) and high-grade glioma ((patient ID: #12 and #19) are shown. Original magnification: ×100

Mentions: We have analyzed the expression of PDGFRα, A2B5, O4 and CD44 as surface markers for glial precursor cells [17], [20]–[24], [27]–[30], and CD133 for stem cells [7], [17] on freshly prepared glioma cells using flow cytometry. In addition, we analyzed the expression of EGFR, which is expressed in immature astrocytes and is critically important for astrocyte development [33]. EGFR is also frequently overexpressed in high-grade glioma cells [34]. Six low-grade and 17 high-grade glioma specimens were analyzed. As shown by representative dot-plot profiles for low-grade glioma specimens (Figure 1), intermingled cells of hematopoietic origin can be distinguished via their CD45 expression. PDGFRα, A2B5, O4, and CD44 were found to be expressed in nearly all CD45 negative cells. The concomitant expression of these progenitor cell markers was observed in nearly all low-grade glioma specimens analyzed (Table 1). These data, combined with the homogenous GFAP staining and the absence of NSE/synaptophysin staining (Table 2 and Figure 2), suggest that low-grade glioma cells are reminiscent of glial-progenitor cells. In 4 out of 6 low-grade glioma specimens, CD133 expression was observed at lower frequencies, but these CD133+ cells are predominantly of blood vessel endothelial origin (see below). The expression of EGFR was found in specimens from 2 of the 6 low-grade gliomas. We also analyzed the expression of CD24. Absence of CD24 expression in combination with positive staining on CD133, or PNA or ABCG2/Bcrp1 was used to enrich primitive neural cells with “stem cell-like” properties [17], [35]–[38]. CD24 expression was detected in 4 out of 6 low-grade gliomas.


Glial progenitor-like phenotype in low-grade glioma and enhanced CD133-expression and neuronal lineage differentiation potential in high-grade glioma.

Rebetz J, Tian D, Persson A, Widegren B, Salford LG, Englund E, Gisselsson D, Fan X - PLoS ONE (2008)

Neuronal marker expression was not detected in low-grade gliomas, but in high-grade gliomas.Staining patterns of GFAP and NSE expression from representative low-glioma (patient ID: #2) and high-grade glioma ((patient ID: #12 and #19) are shown. Original magnification: ×100
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2277459&req=5

pone-0001936-g002: Neuronal marker expression was not detected in low-grade gliomas, but in high-grade gliomas.Staining patterns of GFAP and NSE expression from representative low-glioma (patient ID: #2) and high-grade glioma ((patient ID: #12 and #19) are shown. Original magnification: ×100
Mentions: We have analyzed the expression of PDGFRα, A2B5, O4 and CD44 as surface markers for glial precursor cells [17], [20]–[24], [27]–[30], and CD133 for stem cells [7], [17] on freshly prepared glioma cells using flow cytometry. In addition, we analyzed the expression of EGFR, which is expressed in immature astrocytes and is critically important for astrocyte development [33]. EGFR is also frequently overexpressed in high-grade glioma cells [34]. Six low-grade and 17 high-grade glioma specimens were analyzed. As shown by representative dot-plot profiles for low-grade glioma specimens (Figure 1), intermingled cells of hematopoietic origin can be distinguished via their CD45 expression. PDGFRα, A2B5, O4, and CD44 were found to be expressed in nearly all CD45 negative cells. The concomitant expression of these progenitor cell markers was observed in nearly all low-grade glioma specimens analyzed (Table 1). These data, combined with the homogenous GFAP staining and the absence of NSE/synaptophysin staining (Table 2 and Figure 2), suggest that low-grade glioma cells are reminiscent of glial-progenitor cells. In 4 out of 6 low-grade glioma specimens, CD133 expression was observed at lower frequencies, but these CD133+ cells are predominantly of blood vessel endothelial origin (see below). The expression of EGFR was found in specimens from 2 of the 6 low-grade gliomas. We also analyzed the expression of CD24. Absence of CD24 expression in combination with positive staining on CD133, or PNA or ABCG2/Bcrp1 was used to enrich primitive neural cells with “stem cell-like” properties [17], [35]–[38]. CD24 expression was detected in 4 out of 6 low-grade gliomas.

Bottom Line: In parallel, the expression of glial fibrillary acidic protein (GFAP), synaptophysin and neuron-specific enolase (NSE) was immunohistochemically analyzed in fixed tissue specimens.The rare CD133 expressing cells in low-grade glioma specimens typically co-expressed vessel endothelial marker CD31.The CD133 expression correlates inversely with length of patient survival.

View Article: PubMed Central - PubMed

Affiliation: The Rausing Laboratory, Division of Neurosurgery, Lund University Hospital, Lund, Sweden.

ABSTRACT

Background: While neurosphere- as well as xenograft tumor-initiating cells have been identified in gliomas, the resemblance between glioma cells and neural stem/progenitor cells as well as the prognostic value of stem/progenitor cell marker expression in glioma are poorly clarified.

Methodology/principal findings: Viable glioma cells were characterized for surface marker expression along the glial genesis hierarchy. Six low-grade and 17 high-grade glioma specimens were flow-cytometrically analyzed for markers characteristics of stem cells (CD133); glial progenitors (PDGFRalpha, A2B5, O4, and CD44); and late oligodendrocyte progenitors (O1). In parallel, the expression of glial fibrillary acidic protein (GFAP), synaptophysin and neuron-specific enolase (NSE) was immunohistochemically analyzed in fixed tissue specimens. Irrespective of the grade and morphological diagnosis of gliomas, glioma cells concomitantly expressed PDGFRalpha, A2B5, O4, CD44 and GFAP. In contrast, O1 was weakly expressed in all low-grade and the majority of high-grade glioma specimens analyzed. Co-expression of neuronal markers was observed in all high-grade, but not low-grade, glioma specimens analyzed. The rare CD133 expressing cells in low-grade glioma specimens typically co-expressed vessel endothelial marker CD31. In contrast, distinct CD133 expression profiles in up to 90% of CD45-negative glioma cells were observed in 12 of the 17 high-grade glioma specimens and the majority of these CD133 expressing cells were CD31 negative. The CD133 expression correlates inversely with length of patient survival. Surprisingly, cytogenetic analysis showed that gliomas contained normal and abnormal cell karyotypes with hitherto indistinguishable phenotype.

Conclusions/significance: This study constitutes an important step towards clarification of lineage commitment and differentiation blockage of glioma cells. Our data suggest that glioma cells may resemble expansion of glial lineage progenitor cells with compromised differentiation capacity downstream of A2B5 and O4 expression. The concurrent expression of neuronal markers demonstrates that high-grade glioma cells are endowed with multi-lineage differentiation potential in vivo. Importantly, enhanced CD133 expression marks a poor prognosis in gliomas.

Show MeSH
Related in: MedlinePlus