Introduction
Materials and methods
Patient material selection
Diagnosis | Average age (years) | M/F | Mean OLIG2 score | SD |
---|---|---|---|---|
Glioblastoma, WHO grade IV | 8.7 | 2M/4F | 3 | 0 |
Anaplastic astrocytoma, WHO III | 7.6 | 8M/5F | 2.2 | 0.9 |
Recurrent anaplastic astrocytoma, WHO III | 17 | 2M/0F | 3 | 0 |
Astrocytoma, WHO II | 10.6 | 9M/4F | 2.6 | 0.8 |
Recurrent astrocytoma, WHO II | 11.8 | 2M/2F | 1.4 | 1.1 |
Pilocytic astrocytoma, WHO I | 6.9 | 13M/19F | 3 | 0.18 |
Recurrent pilocytic astrocytoma, WHO I | 10.5 | 2M/0F | 2.5 | 0.7 |
Ependymoma, WHO II | 10.1 | 4M/4F | 0 | 0 |
Recurrent ependymoma, WHO II | 7.5 | 1M/1F | 0.4 | 0.5 |
Anaplastic ependymoma, WHO III | 7.8 | 3M/1F | 0.25 | 0.5 |
Myxopapillary ependymoma | 15 | 1M/1F | 1.5 | 2.1 |
SEGA | 6 | 1M/1F | 0 | 0 |
Tumor 1 | Tumor 2 | P value |
---|---|---|
Anaplastic ependymoma, WHO III | Anaplastic astrocytoma, WHO III | <1 × 10−5
|
Ependymoma, WHO II | Anaplastic astrocytoma, WHO III | <1 × 10−5
|
Pilocytic astrocytoma, WHO I | Anaplastic astrocytoma, WHO III | 0.017 |
Recurrent ependymoma, WHO II | Anaplastic astrocytoma, WHO III | 0.0058 |
SEGA, WHO I | Anaplastic astrocytoma, WHO III | 3 × 10−4
|
Astrocytoma, WHO II | Anaplastic ependymoma, WHO III | <1 × 10−5
|
Glioblastoma, WHO IV | Anaplastic ependymoma, WHO III | <1 × 10−5
|
Pilocytic astrocytoma, WHO I | Anaplastic ependymoma, WHO III | <1 × 10−5
|
Recurrent anaplastic astrocytoma, WHO III | Anaplastic ependymoma, WHO III | 6.6 × 10−5
|
Recurrent pilocytic astrocytoma, WHO I | Anaplastic ependymoma, WHO III | 0.0024 |
Ependymoma, WHO II | Astrocytoma, WHO II | <1 × 10−5
|
SEGA, WHO I | Astrocytoma, WHO II | <1 × 10−5
|
Glioblastoma, WHO IV | Ependymoma, WHO II | <1 × 10−5
|
Pilocytic astrocytoma, WHO I | Ependymoma, WHO II | <1 × 10−5
|
Recurrent anaplastic astrocytoma, WHO III | Ependymoma, WHO II | <1 × 10−5
|
Recurrent astrocytoma, WHO II | Ependymoma, WHO II | 0.017 |
Recurrent pilocytic astrocytoma, WHO I | Ependymoma, WHO II | 7.2 × 10−5
|
Recurrent astrocytoma, WHO II | Glioblastoma, WHO IV | 0.0039 |
Recurrent ependymoma, WHO II | Glioblastoma, WHO IV | 5.1 × 10−5
|
SEGA, WHO I | Glioblastoma, WHO IV | <1 × 10−5
|
Recurrent astrocytoma, WHO II | Pilocytic astrocytoma, WHO I | <1 × 10−5
|
Recurrent ependymoma, WHO II | Pilocytic astrocytoma, WHO I | <1 × 10−5
|
SEGA, WHO I | Pilocytic astrocytoma, WHO I | <1 × 10−5
|
Recurrent ependymoma, WHO II | Pilocytic astrocytoma, WHO I | <1 × 10−5
|
SEGA, WHO I | Recurrent anaplastic astrocytoma, WHO III | 2.1 × 10−3
|
Recurrent pilocytic astrocytoma, WHO I | Recurrent ependymoma, WHO II | 0.031 |
Immunohistochemistry techniques
Immunohistochemistry scoring
Tumor site |
n
| Ave. age | M/F | Mean OLIG2 score | SD |
---|---|---|---|---|---|
Optic nerve | 1 | 3 | 1M/0F | 3 | N/A |
Suprasellar zone | 1 | 3 | 0M/1F | 3 | N/A |
Deep cerebral gray matter | 14 | 6.1 | 8M/6F | 2.7 | 0.6 |
Posterior fossa/cerebellum | 24 | 8.3 | 11M/13F | 2.4 | 1 |
Brainstem | 11 | 7.4 | 6M/5F | 2.5 | 0.82 |
Spinal cord | 8 | 13.1 | 4M/4F | 2 | 1.4 |
Frontal-temporal-parietal lobes | 21 | 9.9 | 13M/8F | 1.8 | 1.2 |
Occipital lobes | 2 | 6 | 2M/0F | 1 | 0 |
Ventricular system | 8 | 9.9 | 4M/4F | 0.6 | 1.1 |
Dual-label OLIG2-Ki67 immunohistochemistry
Genetic analysis of BRAF in pediatric gliomas
In silico analysis of ependymoma and juvenile pilocytic astrocytoma Affymetrix microarray data
Results
Distribution of pediatric brain tumor cases
Distribution of OLIG2 expression in pediatric tumors
Tumor type and anatomic site affecting OLIG2 expression
Cellular proliferation and OLIG2+ expression
Case number | Diagnosis | Percentage of OLIG2+ cells that are also Ki67+ | Percentage of Ki67+ cells that are also OLIG2 positive |
---|---|---|---|
1 | Pilocytic astrocytoma, WHO grade I | 1.6 | 100 |
2 | Pilocytic astrocytoma, WHO grade I | 0 | * |
3 | Pilocytic astrocytoma, WHO grade I | 0 | * |
4 | Pilocytic astrocytoma, WHO grade I | 0.6 | 88.9 |
5 | Pilocytic astrocytoma, WHO grade I | 0.1 | 100 |
6 | Pilocytic astrocytoma, WHO grade I | 0 | * |
7 | Pilocytic astrocytoma, WHO grade I | 5 | 80.3 |
8 | Pilocytic astrocytoma, WHO grade I | 0 | * |
9 | Pilocytic astrocytoma, WHO grade I | 0.5 | 54.5 |
Mean of pilocytic astrocytoma | 0.9 (SEM = 0.5) | 85.0 (SEM = 6.2) | |
1 | Astrocytoma, WHO grade II | 0.1 | 100 |
2 | Astrocytoma, WHO grade II | 0.5 | 100 |
3 | Astrocytoma, WHO grade II | 1.7 | 100 |
4 | Astrocytoma, WHO grade II | 2 | 71.4 |
5 | Astrocytoma, WHO grade II | 0.9 | 100 |
Mean of astrocytoma, grade II | 1.0 (SEM = 0.3) | 94.3 (SEM = 5.7) | |
1 | Anaplastic astro, WHO grade III | 0.2 | 100 |
2 | Anaplastic astro, WHO grade III | 12.4 | 92.9 |
3 | Anaplastic astro, WHO grade III | 2.4 | 86.2 |
4 | Anaplastic astro, WHO grade III | 1 | 94.1 |
Mean anaplastic astrocytoma | 4.9 (SEM = 2.8) | 93.3 (SEM = 2.8) | |
1 | Glioblastoma Multiforme WHO grade IV | 9.2 | 80.4 |
2 | Glioblastoma Multiforme WHO grade IV | 23.6 | 88.7 |
Mean of glioblastoma | 16.3 (SEM = 7.2) | 84.5 (SEM = 4.1) |
BRAF mutation shows no correlation with OLIG2 expressing tumors
Case # | Site | Diagnosis | OLIG2 score | BRAF | Other |
---|---|---|---|---|---|
4035 | Spinal cord | Pilocytic astrocytoma (WHO I) | 3 | K16-B9 | None |
2974 | Cerebellum | Pilocytic astrocytoma (WHO I) | 3 | K16-B11 | None |
2085 | Cerebellum | Pilocytic astrocytoma (WHO I) | 3 | K16-B9 | None |
4035 | Spinal cord | Pilocytic astrocytoma (WHO I) | 3 | K16-B9 | None |
2652 | Deep grey | Astrocytoma (WHO II) | 1 | None | None |
2995 | FTP | Astrocytoma (WHO II) | 3 | None | None |
4282 | Cerebellum | Astrocytoma (WHO II) | 3 | K16-B9 | None |
4825 | FTP | Astrocytoma (WHO II) | 3 | None | None |
7269 | Deep grey | Astrocytoma, anaplastic (WHO III) | 3 | None | None |
1734 | FTP | Astrocytoma, anaplastic (WHO III) | 3 | V600E | NONE |
1762 | IV | Astrocytoma, anaplastic (WHO III) | 1 | None | TP53, 273:R>H |
7124 | FTP | Glioblastoma multiforme | 3 | None | TP53, 172; V>F, PIK3C2B/MDM4 (A), MYC/PVT1(A) |
In silico analysis of ependymoma and astrocytoma tissue microarray datasets
Gene symbol | Ensembl # | logFC |
p
| Probe type |
---|---|---|---|---|
CAM 2
| ENSG00000175161 | 4.90 | 0.04 | _at |
FGF12
| ENSG00000114279 | 4.35 | 0.04 | _at |
BAI-3 | ENSG00000135298 | 4.18 | 0.03 | _at |
SH2
| ENSG00000145147 | −3.83 | 0.009 | _s_at |
Stonin2
| ENSG00000140022 | −4.24 | 0.02 | _at |
RMST
| n/a | −6.54 | 0.02 | _at |
Gene name | Genbank # | Diff exp in OLIG2 null (Y/N) | Mean ExpM-EP (st. err.) | Mean ExpM-PA (st. err.) | ExpM-EP/ExpM-PA |
---|---|---|---|---|---|
Stem cell associated | |||||
SOX2 | NM_003106 | N | 380.23 (13.57) | 150.19(34.43) | 2.53 |
Nestin | NM_006617.1 | N | 254.07 (67.15) | 558.29 (96.20) | 0.46 |
BMI1 | NMJW5180 | Y | 957.30 (62.73) | 497.97 (90.02) | 1.92 |
Anax6 | NM_001155 | Y | 272.54 (34.65) | 486.29 (57.96) | 0.56 |
GFAP | NM_002055 | Y | 11562.63(160.55) | 12983.618(561.08) | 0.89 |
Vimentin | NM_003380 | Y | 5977.26 (262 44) | 6479.32 (2993.85) | 0.92 |
Numb | NM_001005743.1 | Y | 61.27 (14.7) | 46.47 (7.4) | 1.3 |
CCND1 | NM_053056.2 | N | 157.1 (55.68) | 165.18(62.7) | 0.95 |
EGFR-1 | NM_005228 | N | 37.9 (3.37) | 36.99 (4.37) | 1.02 |
Astrocyte associated | |||||
Aqp4 (probe 1) | NM_001650.4 | N | 339 93 (33.06) | 64.16(110.18) | 5.3 |
Aqp4 (probe 2) | NMJW1650.4 | N | 3694(1736.48) | 310.78 (47.43) | 11.89 |
FGF3 | NM_000t42.4 | N | 11.46(1.24) | 10.59(102.42) | 11.46 |
S100b | NM_006272.2 | N | 2362.04 (858.537) | 1827.56(1.72) | 1.29 |
Oligodendrocyte associated | |||||
OLIG1 | NM_138983.2 | Y | 134.82 (14.65) | 2271.04 (328) | 0.06 |
OLIG2 | NM_Q00533.3 | Y | 66.88(19.87) | 663.45(18.77) | 0.1 |
OMB | NM_138983.2 | Y | 56.92 (12.4) | 545.61 (453.16) | 0.1 |
PDGFRa (probe 1) | NM 006206.4 | Y | 384.35 (264.4) | 34.50(133.47) | 1.16 |
PDGFRa (probe 2) | NM 006206.4 | Y | 41.2 (16.984 | 35.4 (3.73) | 1.17 |
MBP (probe 1) | NM_001025081.1 | Y | 50.87(16.49) | 150.01 (9.60) | 0.3 |
MBP (probe 2) | NM_OO1025O81.l | Y | 90.57 (34.76) | 1431.30 (1332.39) | 0.06 |
PLPI | NM_000533.3 | Y | 355(125.54) | 1455.24 (770.10) | 0.24 |