Novel glioblastoma markers with diagnostic and prognostic value identified through transcriptome analysis.
OBJECTIVE
Current methods of classification of astrocytoma based on histopathologic methods are often subjective and less accurate. Although patients with glioblastoma have grave prognosis, significant variability in patient outcome is observed. Therefore, the aim of this study was to identify glioblastoma diagnostic and prognostic markers through microarray analysis.


METHODS
We carried out transcriptome analysis of 25 diffusely infiltrating astrocytoma samples [WHO grade II--diffuse astrocytoma, grade III--anaplastic astrocytoma, and grade IV--glioblastoma (GBM)] using cDNA microarrays containing 18,981 genes. Several of the markers identified were also validated by real-time reverse transcription quantitative PCR and immunohistochemical analysis on an independent set of tumor samples (n = 100). Survival analysis was carried out for two markers on another independent set of retrospective cases (n = 51).


RESULTS
We identified several differentially regulated grade-specific genes. Independent validation by real-time reverse transcription quantitative PCR analysis found growth arrest and DNA-damage-inducible alpha (GADD45alpha) and follistatin-like 1 (FSTL1) to be up-regulated in most GBMs (both primary and secondary), whereas superoxide dismutase 2 and adipocyte enhancer binding protein 1 were up-regulated in the majority of primary GBM. Further, identification of the grade-specific expression of GADD45alpha and FSTL1 by immunohistochemical staining reinforced our findings. Analysis of retrospective GBM cases with known survival data revealed that cytoplasmic overexpression of GADD45alpha conferred better survival while the coexpression of FSTL1 with p53 was associated with poor survival.


CONCLUSIONS
Our study reveals that GADD45alpha and FSTLI are GBM-specific whereas superoxide dismutase 2 and adipocyte enhancer binding protein 1 are primary GBM-specific diagnostic markers. Whereas GADD45alpha overexpression confers a favorable prognosis, FSTL1 overexpression is a hallmark of poor prognosis in GBM patients.
Reddy
Sreekanth P
SP
Department of Molecular Reproduction, Indian Institute of Science, Bangalore, Karnataka, India.
Britto
Ramona
R
Vinnakota
Katyayni
K
Aparna
Hebbar
H
Sreepathi
Hari Kishore
HK
Thota
Balaram
B
Kumari
Arpana
A
Shilpa
B M
BM
Vrinda
M
M
Umesh
Srikantha
S
Samuel
Cini
C
Shetty
Mitesh
M
Tandon
Ashwani
A
Pandey
Paritosh
P
Hegde
Sridevi
S
Hegde
A S
AS
Balasubramaniam
Anandh
A
Chandramouli
B A
BA
Santosh
Vani
V
Kondaiah
Paturu
P
Somasundaram
Kumaravel
K
Rao
M R Satyanarayana
MR
D014408:Biomarkers, Tumor / Q000235:genetics*; D001932:Brain Neoplasms / Q000175:diagnosis* / Q000235:genetics* / Q000401:mortality; D020869:Gene Expression Profiling; D005909:Glioblastoma / Q000175:diagnosis* / Q000235:genetics* / Q000401:mortality; D006801:Humans; D007150:Immunohistochemistry; D053208:Kaplan-Meier Estimate; D020411:Oligonucleotide Array Sequence Analysis; D011379:Prognosis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D014408:Biomarkers, Tumor