cancer genome atlas and functional systems biology wei zhang, ph.d. professor department of...
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Cancer Genome Atlas and Functional Systems
Biology
Wei Zhang, Ph.D.Professor
Department of PathologyDirector
Cancer Genomics Core LaboratoryM. D. Anderson Cancer Center
The 6th Chinese Conference on OncologyMay 21-23, 2010, Shanghai, China
Complexity of CancerComplexity of Cancer
• Cancers have heterogeneous etiology. One patient’s cancer is different from another patient’s cancer.
• Cancers have heterogeneous genetic defects.
• Cancers are results of combinations of multiple genetic and molecular alterations.
Complexity of Human GenomeComplexity of Human Genome
• 30-40,000 genes• 1-10 millions of Single Nucleotide Polymorphisms (SNPs)• 10-20 millions of proteins
• One gene• different spliced mRNAs
• different proteins• different modified forms of proteins
Genomics and Proteomics
Broad-scope, large-scale measurement of gene copy number, gene expression, gene methylation, and protein expression.
Data interpretation or signal processing in pursuit of biological understanding.
DNA RNA
Splice VariantsChIP
• protein/DNA interactions
• transcription• DNA
replication• DNA repair
• splice forms of specific genes
• downstream effects on translation
GX
• high sensitivity measurements of transcription
• correlate results with genomic data
mRNA
Agilent Technologies Microarray Portfolio…
CGH
• chromosomal aberrations
• gene copy number
Copy number
CH3
• methylation patterns
• downstream transcriptional effects
Methylation
Transcription Factors
mRNA isoforms
miRNA
• presence of microRNAs
• knockout analysis
• correlate results with transcription data
RNA interference
What is TCGA?What is TCGA?
• The Cancer Genome Atlas (TCGA). The first phase is a 3-year, 100 million pilot project of the National Cancer Institute (NCI) and the National Human Genome Research Institute (NHGRI) focusing on glioblastoma and ovarian cancer. The second phase will cover 25 major cancer types.
• TCGA Mission: Increase scientific understanding of the molecular basis of cancer and apply this information to improve our ability to diagnose, treat, and prevent cancer.
• TCGA Purpose: Develop a complete “atlas” of all genomic alterations involved in cancer.
TCGA Pilot Project MilestonesTCGA Pilot Project Milestones
Collect/Utilize tumor tissue samples and medical information from cancer patients during treatment.
Catalog and store samples at a centralized facility and send genetic material to research centers involved in the project.
Identify genomic changes associated with cancer in individual patients.
Identify genomic patterns associated with the disease, and use that information to inform cancer diagnosis, treatment, and prevention.
Make information available to scientists as it is produced, to speed treatment and prevention research and help doctors and patients make treatment decisions.
1 2 3 4 5
Graphics credit: The Washington Post, December 14, 2005
How TCGA FunctionsHow TCGA FunctionsData Management, Bioinformatics, and Computational Analysis (GDAC)
An integrated database providing access to all of the information generated by the TCGA pilot project
Technology Development
Throughout the pilot project, technology development will enable improvements to genomic analysis
Cancer Genome Characterization Centers
Technologies to investigate and characterize genes that may be associated with cancer
High-throughput sequencing of genes identified through cancer genome characterization centers
Genome Sequencing Centers
Human Cancer BiospecimenCore Resource
Centralized facility to catalog and store tumor samples, and distribute genetic material to TCGA research centers
Our GDAC Center
• Center for Systems Analysis of the Cancer Regulome
• Directors: Ilya Shmulevich (Institute for Systems Biology); Wei Zhang (M.D. Anderson Cancer Center)
• Bioinformatic researchers at MDACC: Da Yang, Yuexin Liu
• Focuses are on prognosis markers, systems understanding and functional validation
Copy Number Methylation
mRNA expression
BiomarkerSystematic Network
• Prediction Analysis of Microarray
• Top Scoring Pair Algorithm
• Co-occurrence Copy Number Alterations
• Bayesian Network
Tumor Subgroup
• Distinct Dosage Sensitive Expression Patterns
Statistical analysis of mutation significance identified Eight Genes as Significantly Mutated and P53 Mutation Is a Common Event in Primary Glioblastoma.
TCGA., “Comprehensive Genomic Characterization Defines Human Glioblastoma Genes And Core Pathways,” Nature, 455(23), 1061-1068, 2008
Genomic and transcriptional aberration analysis detected New Recurrent Focal Alterations such as Homozygous Deletions involving NF1 and PARK2, and Amplifications of AKT3.
TCGA., “Comprehensive Genomic Characterization Defines Human Glioblastoma Genes And Core Pathways,” Nature, 455(23), 1061-1068, 2008
ILK PI3K
Integrins RTK
IGFBP2
AKTP
PTEN
P
P
P
P
P P
P
Survival
GrowthMetabolism
Migration
Proliferation
Akt Cell Signaling
Kinase domain RDPH
308 473
Akt1
Kinase domain RDPH
309 474
Akt2
Kinase domain RDPH
305 472
Akt3
Chromosome location
14q32
19q13
1q44
Homology 75-84% 90-95% 73-79%
Akt Isoforms
Adapted from Cheng GZ et al. 2008.
Developmental Roles of Akt1/2/3
Akt1 Akt2
Akt3
Neuronal development
Glucosehomeostasis
Cellular growthAngiogenesis
Postnatal Survival
Embryonic Development and Survival
Whole body weight and size
Adapted from Gonzalez and McGraw. Cell Cycle. 2009.
Differential Roles of Akt Isoforms in Cancer
• Differential roles of Akt1 and Akt2 in breast cancer (Hutchinson et al. Can Res. 2004; Arboleda et al. Can Res. 2003)
• Akt2 predominant in ovarian cancer (Noske et al. Cancer Letters. 2007)
• Akt3 important in melanoma (Robertson. Can and Met Rev. 2005)
• Akt activation in glioma correlates with higher tumor grade (Wang et al. Lab Invest. 2004)
Hypothesis
Akt3 is the dominant Akt isoform which preferentially induces Oligodendroglioma
progression
Is there a hierarchy in the ability of Akt isoforms to promote oligodendroglioma development and
progression?
Kristen Holmes
RCAS/tv-a Glial-specific Transgenic Mouse Model
Begemann, M., Uhrbom, L., Rajasekhar, V.K., Fuller, G.N., and E.C. Holland. 2004 Dissecting Gliomagenesis Using Glial-Specific Transgenic Mouse Models. In Zhang, W. and G.N. Fuller (Ed.) Genomic and Molecular Neuro-Oncology. Sudbury: Jones and Bartlett. p233-278
WHO Grade IIOligodendrogliomaNormal Brain
WHO Grade IIIAnaplastic Oligo
Histologic Criteria for Oligodendroglioma Progression
Akt3 Promotes Oligodendroglioma Progression
Tumor Penetrance
Anaplastic OligodendrogliomaGene Combination
PDGFB 81% (35/43) 11% (4/35)
PDGFB / Akt1 77% (42/57) 16% (7/42)
PDGFB / Akt2 39% (11/28) 9% (1/11)
PDGFB / Akt3 100% (35/35) 100% (35/35)
GFP 0% (0/22) 0% (0/22)
AKT1
N O/AO
AKT2
N O/AO
AKT3
N O/AO
Systems BiologySystems BiologySystems biology is an emerging field that aims at system-level understanding of biological systems.
Unlike molecular biology which focus on molecules, such as sequence of nucleotide acids and proteins, systems biology focus on systems that are composed of molecular components. Although systems are composed of matters, the essence of system lies in dynamics and it cannot be described merely by enumerating components of the system. At the same time, it is misleading to believe that only system structure, such as network topologies, is important without paying sufficient attention to diversities and functionalities of components. Both structure of the system and components play indispensable role forming symbiotic state of the system as a whole.
- H Kitano
Probabilistic Boolean network
1. Shmulevich I, Dougherty ER, Kim S, and Zhang W. Probabilistic Boolean network: a rule-based uncertainty model for gene regulatory networks. Bioinformatics 18:261-274, 2002.
2. Shmulevich I, Dougherty ER, and Zhang W. Gene perturbation and intervention in probabilistic Boolean network. Bioinformatics 18:1319-1331, 2002.
3. Shmulevich I, Lahdesmaki H, Dougherty ER, Astola J, Zhang W. Proc. Natl. Acad. Sci. USA 100 (16) 2003.
US Patent # 7,257,563 (Shmulevich, Dougherty, and Zhang)
• Such relationships should also be validated experimentally.
• The networks built from our models should provide valuable theoretical guidance to experiments.
VEGF protein is secreted outside the cells and binds to its receptor on the
endothelial cells to promote their growth.
GRB2GRB2
FGF7FGF7
FSHRFSHR
PTK7PTK7
VEGFVEGF Member of fibroblast growth factor family
Follicle-stimulating hormone receptor
Tyrosine kinase receptor
•The protein products of all four genes are part of signal transduction pathways that involve surface tyrosine kinase receptors.
•These receptors, when activated, recruit a number of adaptor proteins to relay the signal to downstream molecules
•GRB2 is one of the most crucial adaptors that have been identified.
•GRB2 is also a target for cancer intervention because of its link to multiple growth factor signal transduction pathways.
GRB2GRB2
GNB2GNB2
•Molecular studies have demonstrated that activation of protein tyrosine kinase receptor-GRB-2 complex activates ras-MAP kinase-NFB pathway to complete the signal relay from outside the cells to the nucleus.
•GNB2 is a ras family member.
MAP kinase 1MAP kinase 1
c-relc-rel•GNB2 influences MAP
kinase 1, which in turn influences c-rel, an NFB component.
IGFBP-2 in Glioma Progression• Up-regulation of IGFBP2 is one of the most consistent and
distinctive gene expression changes in high-grade gliomas (Fuller et al., 1999)
IGFBP-2 Promotes Motility & Invasion
• IGFBP2 activates expression of invasion enhancing genes and promotes glioma invasion in vitro (Hua Wang et al., Cancer Res., 2003)
MMP2CD10
TIMP-1Fibronectin
Integrin 5Integrin 6
Vinculin
ILK-FAK-PI3K-AKT
Regulated matrix degradation
Guiding migration
Actin stress fiberCytoskeleton reorganization
Migration & survival
Thrombospondin 2
Filamin ABcl-2PUMAp21/WAF1XRCC2
TGF beta RinvasionBradykinin R B2
Centaurin
IGFBP2 Thrombin R
survival
Hua Wang, PhDFirst Prize poster Competition at MDACC Trainee Recognition Day
IGFBP2 is an OncogeneProc Natl Acad Sci USA104(28):11736-41, 2007
First prize in 2007 Trainee Recognition Day at MDACC
American Legion Auxiliary Fellowship
NIH Training grant Pharmacoinformatics fellowship
Sarah Dunlap (now Sarah Smith)
+1-561
c-Myc AP2 NFB
NFNFBB
IGFBP2IGFBP2 •A review of the literature showed that Cazals et al. (1999) indeed demonstrated that NFB activated the IGFBP2 promoter in lung alveolar epithelial cells.
•Higher NFB activity in IGFBP2 overexpressing cells was also found.
NFNFBB
IGFBP2IGFBP2
TNFR2TNFR2
ILKILK•Our real-time PCR data showed that in stable IGFBP2-overexpressing cell lines, IGFBP2 indeed enhances ILK expression.
•In addition, IGFBP2 contains an RGD domain, implying its interaction with integrin molecules.
•ILK is in the integrin signal transduction pathway.
0
1
2
3
4
5
parental c1 c4 c5 c8
Rela
tive
Luci
fera
se A
ctiv
ity
OCT1
p65/p50
p50/p50
IGFBP2 c
lone
Paren
tal
IGFBP2 c
lone
Paren
tal
Non-specific
NH2 COOH
IGF binding domains
5’ TCCAGGGAGCCCCCACCATCCGGGGGGACCCCGAGTGTCATCTCTTCT 3’
R G D 306
GAA
E 306
D306E-IGFBP2 (RGE mutant)
Thyroglobulin type-1 motif(TG domain)
DXXDmotif RGD domain
RGE mutational substitution on IGFBP2
Integrin Linked Kinase
IKKα
IGFBP2
ILK
PI3K
AktP
PH
Receptor Tyrosine Kinase
NFĸBIĸB
IĸB
UU
U
NFĸB
Nucleus
RGD
Target genes
PNFĸB
Ligand
1
Ser 473
GSK3
P
Cyclin D
Proliferation
PIP3
Integrins
Integrin Binding is Required for IGFBP2-mediated Progression
n=500
5
10
15
20
25
30
35
40
45
50
RCAS Combination
Gra
de
III I
nc
ide
nc
e (
%)
n=42 n=32n=50
GFP
PDGFB
PDGFB IGFBP2
PDGFB IGFBP2(RGE)
n=22
IGFPB2 Drives Progression via ILK
0%
5%
10%
15%
20%
25%
30%
35%
40%
45%
50%
RCAS Combination
Gra
de
III I
nc
ide
nc
e (
%)
n=42 n=50 n=28 n=26 n=22
PDGFB
PDGFB ILK
PDGFB IGFBP2
PDGFB ILK-KD
PDGFB ILK-KD IGFBP2
IGFBP2-ILK-AKT pathway
• Critical for cancer development and progression
• Opportunities for drug development
• Systems understanding to cancer and cancer therapeutics
• Predictive instead of reactive medicine
• TCGA is making a major impact on individualized medicine
Future Cancer Biology
Acknowledgment• NIH/NCI GDAC Center grant (Shmulevich/Zhang)• NIH/NCI RO1 CA098503 (Zhang/Fuller)• NIH/NCI NIH R01 CA141432 (Zhang/Fuller)• NIGMS/NIH R01 GM072855 (Shmulevich/Zhang)• Goldhirsh Foundation (Zhang) • James S McDonald Foundation (Zhang)• National Foundation for Cancer Research (Zhang/Hamilton)• NFCR Hope Fund (Zhang)• Anthony Bullock III Research fund (Zhang/Fuller/Sawaya)• The Oreffice Foundation (Zhang/Fuller/Sawaya)• Commonwealth Foundation for Cancer Research (Zhang/Trent)
• NIH/NCI NIH R01 CA098570 (Zhang/Pollock, completed)• NIH R21 GM070600 (Shmulevich/Zhang/Kauffman, completed)• Department of Defense (Zhang, completed)• Texas Higher Education Coordinating Board ARP and ATP grants (Zhang/Fuller, Zhang/Holland, completed)• RGK Foundation (Zhang, completed)
• NIH/NCI P30 CA016672-28 (CCSG)• Tobacco Settlement Fund• Kadoorie Foundation• Goodwin Fund