Post on 13-Feb-2016
description
The c-Met receptor contributes to motility and invasion in high grade STS;
a potential therapeutic target
Sarah E. Myers, Theresa G. Nguyen, Quan-Sheng Zhu, Alexander J.F. Lazar, Raphael E. Pollock, Dina C. Lev
Presented by Gonzalo Lopez, MS
Sarcoma Research LaboratoryMD Anderson Cancer Center
Houston, TX
• Complex karyotype: Leiomyosarcoma MFH/UPS MPNST
• Generally poor prognosis
Sarcomas are heterogeneous
Migration and invasion critical for STS metastasisPrimary tumor
Proliferation/angiogenesis
Migration/invasion Embolism/circulation
TransportArrest in organsAdherence to vessel wallExtravasation
Establishmentof a microenvironment
Proliferation/angiogenesis
Metastasis
What are the molecular drivers of these processes???
Candidate mechanisms: Candidate mechanisms: receptor tyrosine kinases (RTK)receptor tyrosine kinases (RTK)
• RTKs: c-Met, EGFR, VEGFR, c-Kit, etc.RTKs: c-Met, EGFR, VEGFR, c-Kit, etc.• RTKs: central components of signaling networks RTKs: central components of signaling networks
(embryogenesis, cell proliferation, apoptosis)(embryogenesis, cell proliferation, apoptosis)• RTK deregulation: diabetes, developmental defects, RTK deregulation: diabetes, developmental defects,
and cancerand cancer• Activated RTKs: induce pathways of cell migration
and invasion
c-Met receptor: relevant biologyc-Met receptor: relevant biology
• Found in mesenchymal and epithelial tissuesFound in mesenchymal and epithelial tissues
• Ligand HGF (hepatocyte growth factor) Ligand HGF (hepatocyte growth factor)
• Involved in embryogenesis, wound healingInvolved in embryogenesis, wound healing
• Broad range of downstream effectsBroad range of downstream effects
c-Met receptorc-Met receptor
WG Jiang et al. 2005
Experimental goalExperimental goal
To evaluate the impact of c-Met activation and inhibitionon complex karyotype STS migration and invasion
c-Met is expressed in human STS tissue
Leiomyo/UPS TMA
MPNST TMA
Phospho c-Met
HGF
Total c-Met
Phosphorylated c-Met correlates with dismal outcome(HR 2.32; p=0.012)
c-Met is expressed in human STS cell lines
c-Met
SKLM
S1H
T108
0A2
04
RD MES
-SA
SW68
4SW
872
NHF
-actin
c-Met
p c-Met
HT1080 SKLMS1 A204
- + - + - + HGF
Birchmeier,C. Nature Reviews. 2003
HGF activates ERK and AKT HGF activates ERK and AKT
p c-Met
c-Met
β-actin
p AKT
AKT
p ERK
ERK
SKLMS1 A204
- + - + HGF
A204SKLMS1
HGF induces migrationHGF induces migration
HGF - + - +
Avg
. # o
f cel
ls/H
PF
HGF (-)
HGF (+)
SKLMS1
HGF induces invasionHGF induces invasion
A204SKLMS1
HGF - + - +
HGF (-)
HGF (+)
Avg
. # o
f cel
ls/H
PFSKLMS1
• Small molecule inhibitor (PHA-665752, Pfizer)
• Anti c-Met siRNA
Inhibition of c-Met
PHA inhibits c-Met activation and downstream signaling
c-Met
p c-Met
ERK
pERK
pAKT
AKT
- + + + +HGF- - + + +PHA 1 u
M
2.5 uM
5 uM
Mock NT siRNA
- + - + - +
c-Met siRNA effect on downstream signaling
c-Met
p c-Met
ERK
pERK
pAKT
AKT
HGF
DMSO PHA
HGF - + - +
DMSO DMSO + HGF PHA PHA + HGF
c-Met blockade abrogates HGF induced invasion A
vg. #
of c
ells
/HPF
- HGF + HGF
FN1
ITGB5
LAMB1
LAMA5
MMP2
MMP2
GAPDH
- +HGF
LAMB1
LAMA5
ITGB5
FN1
HGF induces migration and invasion genes
ConclusionsConclusions• Human STS samples highly express c-Met, HGF, and Human STS samples highly express c-Met, HGF, and
activated c-Met activated c-Met
• Activated c-Met expression correlates with dismal outcome in Activated c-Met expression correlates with dismal outcome in MPNSTMPNST
• Activation of c-Met receptor on STS inducesActivation of c-Met receptor on STS induces migration and invasionmigration and invasion
• c-Met blockade abrogates these processesc-Met blockade abrogates these processes
Acknowledgements
• Sarcoma Research Laboratory at UTMDACC
Dina C. Lev, MD Raphael E. Pollock, MD/PhD Alex J.F. Lazar, MD/PhD Sarah E. Myers, BS Wenhong Ren, MD Colleagues and Staff