Clinical significance of genetic analysis in glioblastoma treatment
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1 Clinical significance of genetic analysis in glioblastoma treatment Department of Neurosurgery, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan Koji Yoshimoto
2 Can we get prognostic information of GBM patients by genetic analysis in the clinical setting?
3 Outline of this presentation 1. Overview of genetic alteration in GBM 2. Clinical implications of key genetic alterations 3. Genetic analysis bench to bedside 4. Clinical significance of genetic analysis
4 GBM has fewer somatic mutation than other tumors Cancer Genome Landscapes B Vogelstein et al. Science 2013;339:
5 Significance of driver gene mutation What is driver gene mutation? Genetic change that promote cancer progression About 125 genes are listed as Driver genes TP53, PTEN,PIK3CA, VHL, NF1, NF2 etc Cancer Genome Landscapes B Vogelstein et al. Science 2013;339:
6 Key driver gene mutation in GBM Cancer Genome Landscapes B Vogelstein et al. Science 2013;339:
7 Key driver genes in this talk IDH ½ (Isocitrate dehydrogenase) H3F3A (H3 histone, family 3A) BRAF (B-Raf proto-oncogene)
8 IDH1/2 mutations in glioma Hot spot mutation at codon 132 (R132H, R132C, etc) Frequent mutation in low grade glioma Rare mutation in primary GBM TET2 hydroxylases Histone demethylase
9 IDH1 mutation is rare in pgbm 100 % Propor on of muta on Yan (2009) Ichimura (2009) 0 Diffuse astrocytoma (II) Oligodendroglioma (II) Oligoastrocytoma (II) Anaplas c astrocytoma (III) Anaplas c oligoastrocytoma (III) Anaplas c oligodendroglioma (III) Secondary glioblastoma (IV) Primary glioblastoma (IV) Yan H et al. NEJM 2009, Ichimura et al. Neuro-Oncol 2009
10 IDH-mutated GBM show longer survival Yan H et al. NEJM 2009
11 Histone H3.3 (H3F3A) mutation was identified in pediatric GBM Driver mutations in histone H3.3 and chromatin remodelling genes in paediatric glioblastoma Nature 2012 Schwartzentruber et al. Nature 2012 Somatic histone H3 alterations in pediatric diffuse intrinsic pontine gliomas and nonbrainstem glioblastomas Wu et al. Nature Genetics 2012
12 Two critical mutations within the histone tail of H3.3 (K27M, G34R/G34V) Nucleosome Histone variant of H3 H1 H2A H4 H3 H2B H3.1 H3.2 H3.3: active transcription ATRX H3.3 DAXX H3F3A K27 M mutations in thalamic gliomas from young adult patients Aihara et al. Neuro-Oncol 2014
13 Sturmet al. Cancer Cell 2012 Cancer Cell 2012
14 Receptor protein Oncogenic BRAF mutation BRAF mutation is common in melanoma and thyroid cancer V600E mutation is a predominant mutation and therapeutic target V600E BRAF V600E MEK ERK Nature review cancer 2014 Cell proliferation
15 BRAF mutation in glioma Frequent BRAF rearrangement/duplication in pilocytic astrocytoma (KIAA1549:BRAF fusion ) BRAF V600E mutations are frequently detected in PXA, DNT, ganglioglioma BRAF V600E mutations are rare in primary GBM (5%), but V600E mutations might have a survival advantage Dahiya et al, Neuro-Oncol 2014
16 Genetic analysis of GBM from bench to clinic IDH1: R132* (H, C, L, S, G) IDH2: R172* (C, M, K) H3F3A: K27M, G34R/G34V BRAF: V600E Hot spot mutation detection by HRM
17 High Resolution Melting (HRM) PCR-based analysis to identify variations in nucleic acid Nucleic difference is detected in melting curve Clinically relevant technique Wt Mut
18 Representative image of IDH1 analysis Wild Mutant Mutant type needs to be verified by sequencing
19 Procedure of HRM Tumor DNA Real-time PCR system Primer Pair Premix reagent All procedure can be performed within two hours Fast, easy and accurate technique for the detection of hot-spot mutation in driver gene
20 Consecutive 333 glioma cases (Kyushu University Hospital ) Diagnosis Case IDH1 IDH2 BRAF H3F3A Diffuse astrocytoma Oligodendroglioma Oligoastrocytoma Anaplastic astrocytoma Anaplastic oligodendroglioma Anaplastic oligoastrocytoma Glioblastoma Gliosarcoma Astroblastoma Pilocytic astrocytoma PXA Ganglioglioma DNT Others Total
21 IDH1/2 mutation
22 IDH1 mutation is detected in 5/123 pgbm Diagnosis Case IDH1 IDH2 Diffuse astrocytoma Oligodendroglioma Oligoastrocytoma Anaplastic astrocytoma Anaplastic oligodendroglioma Anaplastic oligoastrocytoma Secondary glioblastoma Primary glioblastoma Gliosarcoma Astroblastoma Pilocytic astrocytoma PXA Ganglioglioma DNT Others Total
23 IDH-mutant pgbm-1 Gd Flair Gd Flair 56y F 38y M OS > 610 days OS > 497days (alive) 49y F Middle-aged Less enhanced tumor OS > 850 days
24 IDH-mutant pgbm-2 Gd Flair Young adult Less enhanced tumor Gd 27y F OS: 1086 days 38y F Survival > 964 days (alive)
25 Clinical significance of IDH mutation IDH1/2 mutation in primary GBM is rare IDH1/2 mutation in primary GBM can be a molecular marker of longer survival
26 H3F3A (Histone H3.3) mutation
27 Summary of H3F3A mutation Diagnosis Case H3F3A Diffuse astrocytoma 31 0 Oligodendroglioma 20 0 Oligoastrocytoma 2 0 Anaplastic astrocytoma 30 0 Anaplastic oligodendroglioma 31 0 Anaplastic oligoastrocytoma 7 0 Glioblastoma Gliosarcoma 5 0 Astroblastoma 2 1 Pilocytic astrocytoma 28 0 PXA 4 0 Ganglioglioma 4 0 DNT 6 0 Others 15 0 Total Pediatric: 12 Adult: 3
28 Pediatric supratentorial GBM and Astroblastoma Age Sex Diagnosis H3F3A mutation Surgery Treatment OS(days) 5 F GBM K27M biopsy TMZ+RT M GBM K27M partial TMZ+RT 195(alive) 8 M GBM K27M partial PV+RT M GBM K27M partial TMZ+RT F GBM K27M partial PV+RT M GBM G34R partial TMZ+RT F GBM G34R subtotal PV+RT M Astroblastoma G34R partial TMZ+RT M GBM WT biopsy TMZ+RT F GBM WT subtotal TMZ+RT 2680(alive) PV: Cisplatin and Vincristine
29 GBM cases with K27M mutation Gd Gd Gd FLAIR Gd FLAIR Gd
30 Cases with G34R mutation GBM Gd Gd enhancement(-) T2WI Astroblastoma FLAIR Gd
31 No significant survival difference between K27M and G34R P=0.9
32 8y M OS 240 days H3F3A mutation (-) FLAIR Gd(+) GBM with oligodendroglial component Blood Microsatellite instability phenotype 14/18 markers Tumor
33 3y F H3F3A mutation (-) LOH(-) Pre-ope Small cell GBM GFAP( + ) S100 (+) Olig2 (-) Post-ope TMZ+RT, TMZ (38 cycles) 5y 4m later Tumor recurrence (+) Spinal dissemination (+)
34 3y F H3F3A mutation (-) LOH(-) Tumor recurrence and spinal dissemination Dissemination Disappearance of dissemination FLAI R Gd Removal TMZ 6cycles ICE +IT 4cycles An example of long survival GBM T2WI Gd
35 H3F3A-mutant adult GBM 40y F OS 886days 37y M K27M K27M 37y F OS 526days G34R
36 Clinical significance of H3F3A H3F3A mutation is frequently detected in pediatric population, but can be detected in a subset of deepseated GBM in young adult patients H3F3A mutation can be a molecular marker of poor prognosis especially in pediatric GBM patients
37 BRAF V600E mutation
38 Summary of BRAF mutation Diagnosis Case BRAF Diffuse astrocytoma 31 0 Oligodendroglioma 20 0 Oligoastrocytoma 2 0 Anaplastic astrocytoma 30 0 Anaplastic oligodendroglioma 31 0 Anaplastic oligoastrocytoma 7 0 Glioblastoma Gliosarcoma 5 0 Astroblastoma 2 1 Pilocytic astrocytoma 28 4 PXA 4 3 Ganglioglioma 4 4 DNT 6 1 Others 15 0 Total
39 BRAF mut GBM (66y M) FLAIR Gd(+)T1WI Tumor removal followed by TMZ + RT Died form pneumonia No tumor recurrence OS 1366days
40 BRAF mut GBM (38y F) FLAIR CT/ Gd(+)T1WI Onset of epilepsy Followed for 12 years Tumro removal TMZ+RTx PFS 255days OS 444days Malignant transformation of ganglioglioma?
41 Clinical significance of BRAF mutation BRAF-V600Emutation is rare in GBM BRAF-V600Emutation in GBM can be a molecular marker of longer survival
42 Acknowledgements These works are supported by the member of department of neurosurgery at Kyushu University Masahiro Mizoguchi Ryusuke Hatae Yojiro Akagi Hideki Murata Daisuke Kuga Nobuhiro Hata Tetsuro Sayama Prof. Koji Iihara
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