the met oncogene: from molecular biology to clinical trials paolo m. comoglio, md
DESCRIPTION
The MET oncogene: From molecular biology to Clinical Trials Paolo M. Comoglio, MD Institute for Cancer Research @ Candiolo, University of Turin, Medical School, Italy . P. P. P. P. MET IS A ‘MASTER GENE’ IN THE CONTROL OF INVASIVE GROWTH. Sema domain. HGF (Scatter Factor). 500 AA. - PowerPoint PPT PresentationTRANSCRIPT
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The MET oncogene:
From molecular biology to Clinical TrialsPaolo M. Comoglio, MD
Institute for Cancer Research @ Candiolo,University of Turin, Medical School, Italy
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MET IS A ‘MASTER GENE’ IN THE CONTROL OF INVASIVE GROWTH
S-SKringles Protease-
like
Sema domain
G-P richIg like domains
500 AA
400 AA
Tyrosine Kinase
Met(HGF Receptor)
PP
PP
MRS (PSI)
HGF (Scatter Factor)
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P
GAB1
P
P
GRB2RAS
P
P
CD44v6
MET
Plexin B1
Integrin 6 4
SHP2
SRC
Sustained MAP and PI3 kinase activation (invasion / apoptosis protection)
GRB2
Transient MAP kinaseactivation (proliferation)
Link to cytoskeleton
PSTAT
PLC
PI3Kp85
PI3Kp85
CRKL
C.Boccaccio and PM.Comoglio. Nature Rev. Cancer.2006, 6: 637-645.
Cell-Polarity and Morphogenesis
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Xenopatient
Xenosphere(cancer stem cells)
Human metastaticcolorectal cancer
Spheropatient
MET IS A FUNCTIONAL MARKER OF
CANCER STEM CELLS
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Xenopatients, Xenospheres and Spheropatients
Xenopatient Xenosphere(cancer stem cells)
Spheropatient(Secondary xenopatient)
C. Boccaccio et al., in preparation
Mutations retained and passed onResponse to targeted drugs retained and passed on
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MET inhibitionpromotes differentiation of CRC colospheres
Control
JNJ inhib.
Cetuximab
Fibroblast conditioned medium0% serum
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ACTIVATION OF THE MET-DRIVEN ‘INVASIVE GROWTH’ PROGRAMME
GENERATES AN AGGRESSIVE PHENOTYPE
• ONCOGENE ‘EXPEDIENCE’• ONCOGENE ‘ADDICTION’
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THE CONCEPT OF ‘ONCOGENE EXPEDIENCE ‘
• Met can promote invasion and metastasis independent of the genetic lesions that caused tumour onset
• Met activation is a ‘stress’ response to unfavourable microenvironment (hypoxia, radiations , inflammatory cytokines)
• Met activation occurs mainly through transcriptional upregulation
• Met activation is an expedience that provides a pro-survival and invasive advantage
Trusolino and Comoglio., Nature Rev. Drug Discov, 2008Nature Rev. Mol. Cell. Biol. 2010
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The MET promoter
Proteasomedegradation
Cellular [O2]
HIF-1(constitutive)
Nucleus
HIF-1(regulated) -OH
pVHL
Cytoplasm
HIF Proline Hydroxylase
HIF-1
Hypoxia promotes invasive growthby transcriptional activation of the MET Oncogene
P (-2619) S (-345) S (+89)
HRE-1 HRE-2 asHRE-1
AP-1
HRE-4
STARTA (-253)
asHRE-2/HRE-3
S (-411) A (-32)
S (-295)
A (+353)
HRE-5
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met mRNA
21%
O2
3% O
2
CoC
l 2
Con
trol
28S
MET
HIF-1a
Pennacchietti, Michieli et al. , Cancer Cell 3: 347 (2003)
Hypoxia promotes invasive growthby transcriptional activation of the MET Oncogene
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• In a limited number of cancer types, genetic lesions of Met are selected along the tumour natural history to become the driving force that maintains the transformed phenotype
• Met activation is the consequence of a fixed and transmissible genetic alteration; therefore, it is characterized by chronic firing and high steady-state signalling
THE CONCEPT OF ‘ONCOGENE ADDICTION ‘
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• Amplification (e.g. gastric ca, NSCLCs and CRC mts resistant to targeted therapy)
• Point mutations - hereditary and sporadic carcinomas (e.g. kidney, hepatocellular, head
& neck ca.- early metastatic cancers of unknown primary origin (CUPs)
‘Oncogenic addiction’ to MET in tumourscan be achieved by:
Trusolino and Comoglio, Nature Rev. Drug Discov. 2008
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TARGET THERAPIES AGAINST MET
• An adjuvant approach for a vast number of tumours:Overexpression: targeting ‘expedience’(almost all solid tumors expressing MET)- Endpoints: PFS, OS
• A front-line intervention for a limited fraction ofgenotype-specific tumours:Amplification, mutations : targeting ‘addiction’ (e.g. gastric ca., NSCLC, gefitinib-resistant NSCLCs)- Endpoints: Remission
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THERAPEUTIC INACTIVATION OF THE MET ONCOGENECAN BE ACHIEVED BY:
1. Small molecules inhibiting the Tyrosine Kinase
2. Monoclonal Antibodies
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RESPONSE TO MET INHIBITORS CORRELATES WITH GENE AMPLIFICATION
PHA-665752PHA-665752
97 %EXPEDIENCE
5.2NCI-H1993
5.6SNU5
6.3HS746T
6.1GTL-16
6MKN-45
5.8EBC-1
MET copy N°Cell Line
Hypoxia
(or HGF)
< 3 %ADDICTION
200 cancer l. tested
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Clinical trials for MET inhibitors
• Inhibition of growth (reduction of tumor mass) is expected only in cancers “addicted” to MET
• Genetic lesions are expected in ~ 3-4 % of epithelial cancers
• It is mandatory to identify patiens with cancers “addicted” to MET before recruitment
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111In DTPA-DN30 tumor uptake at 2h post injectionSPEC-CT scan
EBC1A2780
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Bardelli. et al. Cancer Discovery 2013
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Detection of MET-amplificationby Liquid Biopsy
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Xenopatient-derived cancer cells display MET amplification and are sensitive to Met targeted drugs
00,20,40,60,8
11,21,4
NT 10 50 500 10 50 500
cell
viab
ility
%
JNJ CRIZOTINIB
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Therapy with MET antibody
MV-DN30 Monoclonal AntibodyMonovalent, Chimeric, Stabilized
• Recombinant, properly assembled and PEGylated• Bind Met (IP4) with high affinity (Kd= 0,116 nM)• Down-regulate the Met receptor from the cell surface• Induce shedding of the extracellular domain (“decoy”)
O
OHn
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Adam 10
p55
p125
p50Metp175
Inactive receptor
heterodimerS -S
Ligand neutralization
p55
p125
p50Metp175
Inactive receptor
heterodimer
Ligand neutralization
Shedding
Γ- secretase
DecoyInhibitoryeffects
Proteasome degradation
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Exogenous administration of anti-MET DN-30 inhibits tumor growth and prevent metastasis
Mab DN30
Human Breast Carcinoma, transplanted in Athymic nu/nu mice
Petrelli, A. et al., Proc. Natl.Acad Sci. US: 2006, 28, 5090-5095
Irrelevant Mab
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0
200
400
600
800
1000
1200
1400
1600
1800
2000
2200
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28
Days after injection
Tum
or v
olum
e (m
m3)
.
NTCDDP5FUMV DN30CDDP + MVDN305FU + MVDN30
CHEMOTHERAPY (CDDP, 5FU) vs MET ANTIBODY TREATMENT OF EBC1
- Xenotransplants-
Days after injection
Tum
or V
olum
e
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MECHANISMS OF ACQUIRED RESISTANCETO MET KINASE INHIBITORS
• MET amplification
• Activating point mutations
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Amplification of MET contributes to acquired resistance to MET kinase inhibitors
GTL16 wt
Chromosome 7 centromereMET amplicon marker
Wt PHA resistant (150 nM)
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WT
1 2 3 4 5 6 7 8 9
a-pMet
a-Met
PHA 250 nM
Y1349
Y1356
PP
PP
PP S985
Y1003
D1228H (Kit)D1228N (Kit)Y1230CY1230HM250T (Ret)
M1131TV1188LL1196VV1220I
Some MET mutations confer resistance to MET inhibitors
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EBC1 wt EBC1 PHA Resistant
FAb DN30 treatment overcomes resistanceto the MET kinase Inhibitor PHA-665752
Viab
ility
in 5
0 nM
PHA
EBC1Wild Type
EBC1PHA Resistant
DN30 Fab 0.4 mM
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MET and Invasive Growth: State of the art:
1. Scatter factors and the receptors of the MET oncogene family (RON, ROR) are key regulators of the Invasive Growth program in cancer stem cells.
2. The program is activated by MET over expression, induced by unfavourable micro-environmental conditions, such as hypoxia or ionizing radiations: (“Oncogene Expedience”).
3. The invasive growth program is constitutively activated in some cancers, by MET amplification, mutations or autocrine loops (“Oncogene Addiction”).
4. The invasive growth program driven by the oncogene MET is a good target for therapy.
5. Resistance may occur, that may be overcome by combined treatment with s.m. inhibitors and antibodies
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Plexins L.TamagnoneE. GiraudoG. Serini
Met AddictionL.TrusolinoA. BertottiF. Galimi
Met GeneticsA. BardelliM.F. Di Renzo
Met & HypoxiaP. MichieliS. Pennacchietti C. Basilico
Met SignatureE. MedicoMet BiologyL. Lanzetti
Met & Stem cellsC. BoccaccioF. De BaccoP. LuraghiG. Reato
Met and resistanceS. GiordanoS. CorsoA. PetrelliC. Migliore
IRCCExperimental Clinical Molecular Oncology
Met Gene TherapyE. VignaS. CignettoR. Albano
Ron & RORS. BenvenutiA. GentileL. LazzariA. Arnesano
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IRCC: Institute for Cancer Research @ Candiolo
“Today Science, Tomorrow Medicine”