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 metastasis

Primary tumorProliferation/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

SKL

MS1

HT

1080

A20

4

RD

ME

S-SA

SW68

4

SW87

2

NH

F

-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 ce

lls/

HP

F

HGF (-)

HGF (+)

SKLMS1

HGF induces invasionHGF induces invasion

A204SKLMS1

HGF - + - +

HGF (-)

HGF (+)

Avg

. # o

f ce

lls/

HP

FSKLMS1

• 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

uM 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

cell

s/H

PF

- 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 Activated c-Met expression correlates with dismal outcome in MPNSTin MPNST

• 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