growth factor pathway activation in chordoma
DESCRIPTION
Growth Factor Pathway Activation in Chordoma. Carolyn Hoban; Dafydd Thomas; David Lucas; Laurence Baker, University of Michigan, Departments of Internal Medicine, Division of Oncology/Hematology; And Pathology, Ann Arbor, Michigan. Chordoma. Backgound: - PowerPoint PPT PresentationTRANSCRIPT
CTOS 11.13.08CTOS 11.13.08
Growth Factor Pathway Activation
in Chordoma
Carolyn Hoban;
Dafydd Thomas; David Lucas; Laurence Baker,
University of Michigan, Departments of Internal Medicine,Division of Oncology/Hematology;
And Pathology,Ann Arbor, Michigan
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Backgound:
Tumor microenvironment of bone and nervous system
Usually slow growing tumors believed to arise from remnants of notochordal tissues
Challenging to treat with surgery, radiation and/or chemotherapy regimens due to location of the tumor.
Frequently recur and may have distant metastasis in late stage of disease. Factors regulating recurrence have not
been determined.
Chordoma
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Translational focus
To determine expression levels and activation of key growth factor pathways immunohistochemically using quantitative methods.
Retrospective analysis of routinely processed paraffin sections of chordoma clinical specimens.
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34 cases of Chordoma obtained from UM pathology archives with IRB approval
3 tumor cores /pt /TMA Diagnostic confirmation upon review by pathologists (Lucas)M:F = 14:14; Age: avg. 52 (Range 11-81)Anatomic location:
Clival (n=11); Sacral (n=9)Primary and recurrent tumors
Chordoma Clinical Specimens
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Low to intermediate-grade tumor,resembling notochord Clusters of large polymorphous cells in myxoid matrix
Pleomorphic & hyperchromatic nucleiMucin filled vacuoles ‘physaliferous’
Positive IHC for Cytokeratin19, S100, EMA
Chordoma Histology
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IHC: Identification of activated pathways & drug targets
TARGET DRUG (example)
Brachyury na
cKIT imatinib, dasatanib
EGFR erlotinib…
EGFR VIII HKI-272
HER2 trastusumab, panitumimib
IGF1R R1507, A12..
ihh GDC0449
mTOR rapalogs
PDGF-R alpha TKI
PDGF-R beta TKI, dasatanib
RET sutent, sorafenib, zd6474
shh GDC0449
TGFb-R2 siTGFBRinh
Topo I 9-NC
Topo II alpha doxorubicin
Topo II beta
DNA cisplatin
Brachyury* [Chordoma marker]
RTK:
HER family
PDGFR and ,
IGF1R,
c-Ret,
c-kit,
c-src,
mTOR, AKT, and MAPK
• 2008 Tirabosco: Am J Surg Pathol, • 2005 Henderson Genome Biology• 2006 Vujovic J. Paht.
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TMA 8387
TMA 6971
TMA 6971 TMA 6983TMA 6974
Chordoma TMA: IHC scores
Brachyury: highhigh frequency 100%
high intensity 3+
c-kit: neg/ low
HER family1. EGFR mod2. EGFRVIII neg3. Her2: neg/ low
Fasig et al. ckit low (33%); EGFR (67%)Weinberger et al. Her2 focal staining
1 2 3
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IHC scores: GF receptor profile
PDGF_R: high
c-ret: high(GDNF pathway)
IGF1R Phospho-tyr IGF1R
TMA 6977
PDGF-RPDGF-R
IGF1R pY1147-IGF1R
TMA 6985
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Automated Quantitative Analysis (AQUA)
Advantages:Automated, high-throughput, Objective (r/o intra/inter-observer concordance; lesion heterogeneity)Fluorescence-based Ab used in IHC
Multiplex:Antigen of interestTumor- and stromal-specific markerSubcellular location (nuclei, cytoplasm,PM)Automated scoring and data analysis Activation index (phospho:total GF-receptor)
Quantitative IHC
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Stroma
Tumor
Necrosis
Cytokeratin Cy3
Nuclear Component
Tumor Mask
AQUA analysis
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0200400600800
1000120014001600
c-kitEGFREGFRVIIIHer2IGF1Rc-RETPDGFR-aPDGFR-b
IGF1R, c-ret, PDGFR pathways are activated
AQUA score
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From Clinical evidence to Preclinical models
TARGET DRUG (example)
Brachyury na
cKIT imatinib, dasatanib
EGFR erlotinib…
EGFR VIII HKI-272
HER2 trastusumab, panitumimib
IGF1R R1507, A12..
ihh GDC0449
mTOR rapalogs
PDGF-R alpha TKI
PDGF-R beta TKI, dasatanib
RET sutent, sorafenib, zd6474
shh GDC0449
TGFb-R2 siTGFBRinh
Topo I 9-NC
Topo II alpha doxorubicin
Topo II beta
DNA cisplatin
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In vitro models (chordoma cell lines) U-CH1 (Scheil et al 2001)
[courtesy of Chordoma Foundation, Josh Sommer; Mike Kelly, Duke Univ)
Morphological features: physaliferous, round nuclei, mucinous, (BrU,CK, S100 & EMA positive)
Other cell lines in development
In vivo models (chordoma/ microenvironment) UCH-1 cells with stable expression of luciferaseDeveloping BLI- Xenograft lines (passage in SCID mice) Plan to evaluate drugs that inhibit GF pathways in vivo
Preclinical models of chordoma
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Used data obtained from clinical specimens to build our experimental models:
Rank cell lines with similar features to clinical specimens (molecular markers, immunopositive GF pathway activation)
Test drug combinations to optimally inhibit key pathways, in vitro (cell kill) and in vivo (OS)
Support nomination of optimal combinations of drugs into clinical research
Translation Clinic-lab-Clinic
Conclusion
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The team:University of Michigan Multidisciplinary Sarcoma groupCollaboratorsPatients & friends
(the village)
Financial support from Stefan L. Harris Fund
Contact: [email protected]
Thank you