Glioblastoma Multiforme
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1 Glioblastoma Multiforme Highly malignant, invasive, difficult-to-treat primary brain tumor" " Frequency: 9,000 cases/year (peak age, years)" " Recurrence: rapid growth; size may double every 10 days" " Median survival: ~ 1 year"
2 Survival of adult patients with glioblastoma multiforme" Kaplan-Meier Survival Curves Survival % Months from Surgery
3 Pediatric Brain Tumors" Frequency : 3000 cases/year"
4 Pediatric brainstem glioma" Brainstem location represents 8-15% of all brain tumors in the pediatric population" Usually inoperable tumors because of the particular location in the brain "
5 Survival for Children with Diffuse Pontine Gliomas (CCG 9941)" J Clin Oncol 20: , 2002
6 Tumor cells multiply which results in growth"
7 Normal growth is controlled"
8 Why do tumor cells grow?" Tumor cells receive the instructions to grow but are insensitive to instructions to stop"
9 Propagation of neural stem cells" Blue: nucleus" Green: nestin" Nestin: marker of stem cells"
10 Differentiation of neural stem cells " in neurons and glia" Blue: nucleus" Green: GFAP" astrocytes" Red: β-iiitub" neurons"
11 Brain development requires a controlled switch from proliferation to differentiation?"?" Stem Cells Differentiated " neurons
12 Neurogenesis and Brain tumors " Disruption of pathways essential for neurogenesis have been implicated in childhood and adult brain cancers, for which immature progenitor cells have been proposed as cells-of-origin"
13 Id proteins: inhibitors of differentiation" Undifferentiated state Differentiated state High growth potential High amounts of Id proteins Low growth potential Low amounts of Id proteins Iavarone and Lasorella, 2003
14 Id proteins are antagonists " of transcription factors Ø No Id proteins bhlh heterodimer CANNTG E-box Activation of" transcription" and differentiation Ø Id proteins in functional excess Id2 Id2 Id2 Id2 CANNTG E-box Inhibition of" transcription and" block of differentiation
15 The Rb-Id2-bHLH pathway in pediatric tumors Myc IGF EWS-Ets Rb Id2 bhlh p57 Kip2 G 1 S M G 2
16 Normal cells Cancer cells Cancer cells invade normal tissues Rb Id2 Rb Id2 Wild type Rb Id2 inactive Mutant Rb Id2 hyperactive
17 Id2 loss impairs tumor growth and " angiogenesis in tumors from Rb+/- mice" Rb+/- Id2-/-;Rb+/- Microfil-orange " Latex perfusion " Pecam-1"
18 Id proteins are coexpressed with HIF1α in human glioblastoma" Id1" Id2" HIF1α
19 Id2 overexpression in neuroblastoma" is associated with reduced survival Survival (percentage) 100" 80" 60" 40" 20" Overall study population Id2" negative, n=18" Id2"positive, n=29" Time (months after diagnosis) P = " 0" 20" 40" 60" 80" 100" 120"
20 Id proteins involved in all processes associated with development of neural tumors VEGF Signaling Integrins, MMP2 Id Metalloproteinases Angiogenesis Tissue Invasion Anaplasia Lineage Specific bhlh Id Proliferation Rb, bhlh, Ets, Pax
21 THE FUTURE : Anti Id2 therapeutics" Growth arrest Differentiation Id2 Id2 Id2 Id2 Id2 Id2 Id2 Id2 Id2 Increased cell death Inhibition of angiogenesis
22 Developmental lineages derived from the neural crest and the genesis of neuroblastoma"
23 Underlying challenge: how to control stem cells?"?"
24 Control brain tumor/neural stem cell behavior Glia Stem cell N-Myc Huwe1 N-Myc Neuron
25 Loss of Huwe1 expands " the neural stem cell population" Pax6/DAPI Huwe1 F/Y! Huwe1 F/Y Nes! Pax6/DAPI IZ" IZ" SVZ" SVZ"
26 Loss of Huwe1 impairs" neural stem cell differentiation" Nestin/DAPI" Huwe1 F/Y! Nestin/DAPI" Huwe1 F/Y Nes! 50 µm βiii-tubulin/dapi" βiii-tubulin/dapi"
27 Focal deletions and decreased expression of Huwe1 in GBM TCGA Oncomine Normal brain n=23 P-value: 9.3E-10 GBM n=77
28 Expression of Huwe1 is lost in primary neuroblastomas displaying accumulation of N-Myc protein" Huwe1 N-Myc #3524 #3629
29 Neural stem cell Growth factors N-Myc N-Myc Growth factors Huwe1 Lineage commitment Differentiation Cell cycle arrest Tumor stem cell N-Myc Huwe1 N-Myc Tumor growth CD2 p27 Growth arrest Maturation
30
31 Malignant gliomas invade the normal brain
32 The mesenchymal signature of high-grade glioma" Unsupervised clustering of 76 high grade tumors by expression of 108 genes that are positively or negatively associated with survival reveals 3 tumors classes - Proneural (PN), Mesenchymal (Mes) and Proliferative (Prolif)." Malignant gliomas belonging to the mesenchymal sub-class express genes linked to the most aggressive proper9es of glioblastoma (migra9on, invasion and angiogenesis) and mark the worst clinical outcome.
33 The mesenchymal network of six major hubs " of transcription factors in high-grade gliomas" Mesenchymal genes Activator" Repressor"
34 STAT3 and C/EBPβ inhibit neuronal differentiation and induce mesenchymal transformation in neural stem cells" Tau SMA Tau/SMA/Dapi" FN1" Ctgf" Vector" Stat3C/CEBPβ E" 5" 10" E" 5" 10" Vector" Olig2! βiiitubulin" βactin" Stat3C/ CEBPβ Vector" Stat3C/ CEBPβ Stat3C/ CEBPβ" Untreated" 20 days"
35 Knockdown of Stat3 and C/EBPβ cooperates to inhibit tumor cell invasion and angiogenesis" shctr" shstat3" shc/ebpβ shstat3+shc/ebpβ shctr" Vimen9n shstat3+shc/ebpβ Vimen9n T T B B µm CD31 CD31
36 Loss of Stat3 and C/EBPβ in human glioma cells inhibits tumorigenesis in the mouse brain" Human glioma-shctr" Human glioma-shstat3+shc/ebpβ 100" Cumulative Survival" 80" 60" 40" 20" ** shstat3+shc/ebpβ shctr" 0" 40" 60" 80" 100" 120" Days post-injection"
37 The combined expression of Stat3 and C/EBPβ correlates with the poorest outcome of glioma patients" Negative Stat3 + C/EBPβ " Positive Stat3 + C/EBPβ "
38 Glioblastoma From systems biology to prognosis to personalized therapy" Mesenchymal Signature Of High Grade Glioma ARACNe Regulatory Network CEBPB STAT3 bhlh-b2 FOSL2 RUNX1 Stat3 and C/EBPβ are Master Regulators Stat3 and C/EBPβ are Transforming Oncogenes of Neural Stem Cells Stat3 and C/EBPβ are Predictors of Negative Clinical Outcome
39
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