www.IRCC.it Oncogene Addiction and Expedience: The Met paradigm - - PowerPoint PPT Presentation

Oncogene ‘Addiction’ and ‘Expedience: The Met paradigm Paolo M. Comoglio MD, pcomoglio@gmail.com www.IRCC.it

Background

• Cancer is a disease of genes
• It is sustained by ‘Stem Cells’
• Can be treated, if the responsible gene(s) are

identified (‘drivers’)

• If targeted drugs are available
• If ‘resistance’ can be prevented / overcomed

The ‘MET’ paradigm

• Cancer is a disease of genes: MET is a potent oncogene
• It is sustained by ‘Stem Cells’: MET is expresseed in stem

cells

• Can be treated if the responsible gene(s) (‘drivers’) are

identified: MET is a driver oncogene

• If targeted drugs are available: good MET kinase

inhibitors and antibodies available

• If ‘resistance’ can be prevented / overcome: possible

Aberrant activity of the MET oncogene in human cancers

Oncogene addiction (DNA) Oncogene expedience (mRNA) METwt gene overexpression MET genetic alteration FREQUENT Transcriptional induction:

• hypoxia
• ionizing radiation

RARE

• Chromosom. rearr.

(TPR-MET, e.g. gastric K) Gene amplification (e.g. gastric and esoph. K; Gefitinib-resistant NSCL) Point mutations (e.g. Papillary kidney K; “cancer of unknown primary site”)

• MET genetic lesions behave as ‘drivers’, being

required and sufficient to initiate and sustain neoplastic transformation (primary ‘addiction’).

• MET lesions are selected during the Darwinian

evolution of cancer, under therapeutic pressure (secondary ‘addiction’)

• Identification of MET as driver and tailoring specific

drugs may result in efficient ‘precision therapy’. ‘Oncogene addiction’:

‘Primary oncogene addiction’: the response matches exactly Met amplification

J&J 605 Specific MET kinase inhibitor

5.2 NCI-H1993 5.6 SNU5 6.3 HS746T 6.1 GTL-16 6 MKN-45 5.8 EBC-1 MET copy N° Cell Line < 3 % ADDICTION

197 cancer cell lines tested

Diagnosis should be ‘molecular’ (The ‘precision medicine’ approach)

• Cancer is a disease that develops in an organ
• It is not a disease of the organ
• A given oncogene (e.g. MET) may hit different organs
• Cancers in different organs may respond to the same

MET-targeted drug

Plasma Tumor

Patient DM – Chromosome 17

(A.Bardelli et al. 2013)

The ‘precision medicine’ approach

(Liquid Biopsy followed by next gen. sequencing)

MET MET

Lung Ca MET ampl. 12x 2h post injection 2h post injection

SPEC-CT scan Imaging the MET oncogene amplification by

111In DTPA-DN30 antibody

Ovary Ca MET wt

500 1000 1500 2000 2500 3000

• 40
• 30
• 20
• 10

10 20 30 40 50

Tumor Volume (mm3) Days

Vehicle Cetuximab Crizotinib Cetuximab+Crizotinib JNJ-38877605 Cetuximab+JNJ- 38877605 Xenopatient M162

Response to specific inhibitors by a ‘xenopatient’ bearing a MET amplified colorectal Ca.

L.Trusolino et al, 2014

A B

Patient #1: before Pmab Patient #1: after Pmab Patient #2: before Pmab Patient #2: after Pmab

C

Patient #3: before Cmab Patient #3: after Cmab 40X 40X Met IHC 60X 60X 60X MET CEP7 MET CEP7 MET CEP7 60X 60X 60X MET CEP7 MET CEP7 MET CEP7 40X 40X 40X Met IHC Met IHC 40X

Bardelli A et al., Cancer Discovery, 2013

MET amplification is associated with secondary addiction in anti-EGFR resistant patients

• Bardelli. et al. Cancer Discovery 2013

MET amplification is associated with secondary addiction in anti-EGFR resistant patients

• Some wild-type oncogenes, including MET, are

activated in cancer cells as an adaptive response to adverse microenvironmental conditions (e.g. hypoxia, nutrient starvation, or ionizing radiation), favour tumor progression and confer therapeutic resistance (‘expedience’). ‘Oncogene expedience’:

Ionizing radiations activate the MET oncogene

Ionizing radiations

ROS

DNA damage (...) MET

P

NFB RelA p50 IKKa-b Nemo IB ATM

P P

Radioresistance MET overexpression

P P

De Bacco et al., J Natl Cancer Inst.; 2011.

MET ‘Adjuvant Therapy’ enhances the response to radiation therapy of human GBM xenografts

De Bacco et al., 2015, in press.

MET ‘Adjuvant Therapy’ enhances the response to radiation therapy of human GBM xenografts

Epifluorences of human GBM xenografts transduced with GFP Vehicle JNJ anti-MET 2 Gy/day x 3 Combo (*)

(*) JNJ at day 0 followed by daily administration of 25 mg/g for 30 days, De Bacco et al., 2015, in press.

The so called ‘Cancer Stem cells’

• Cancer develops from transformation of a

stem/progenitor cell into a ‘cancer stem cell’.

• Conventional anti-neoplastic drugs efficiently kill the

‘mature’ cancer cells, but not cancer stem cells

• Often cancer recurs from its roots

• Some wild-type oncogenes (such as MET), inherited

from the cell of origin (a normal stem/progenitor), govern an essential signaling circuit that sustains the inherent self-renewing, self-preserving and malignant phenotype of the cancer stem cell (‘inherence’) . Oncogene ‘Inherence’:

Efficient anti-MET drugs are available (with some problems)

• Small molecule specific kinase inhibitors

e.g. Crizotinib, J&J 605, …. (problem: ‘rebound effect’)

• Antibodies agains the HGF ligand binding site

(problem: MET activation in most cancers is ‘ligand-independent)

A non-conventional MET antibody

MV-DN30 Monoclonal Antibody Monovalent, Humanized, Chimeric, Stabilized

• Recombinant Fab, properly assembled and PEGylated
• Binds Met with high affinity (Kd= 0,116 nM)
• Binds MET at the IP4 domain, outside the HGF binding site
• Down-regulate the Met receptor from the cell surface
• Induce shedding of the extracellular domain (generating a “decoy”)

O OH n

p125 p125 p50 Met p175 Inactive Receptor heterodimer Ligand Ligand neutralization Adam 10 Shedding Γ- secretase Decoy Inhibitory effects Proteasome degradation

Reviewed by: Vigna E. and Comoglio P.M., Oncogene.34:1883-89; (2015)

Non-conventional reponse to MV-DN30 Monoclonal Antibody

MV-DN30 p55

‘Gene Therapy’ with MET antibody

Gene transfer into the tumor: Cancer cells produce the Monoclonal Antibody Bi-cistronic Lentiviral vector carrying the cDNA for H and L chains of MV-DN30 (Tet-inducible promoter)

An orthotopic mouse model of human GBM

LV- MvDN-30 vector

Gene therapy with LV(Tet)-DN30 FAb (U87 Glioblastoma )

10 20 30 40 50 60 70 80 90 100 10 20 30 40 50 60 70 80 90 100 110

Time (days) % tumor-free animals . no dox ( Fab-) dox + (Fab+)

Vigna et al., Cancer Res. ;68:9176-83, 2008

Mechanisms of acquired resistance To met kinase inhibitors

• MET amplification
• Activating point mutations
• Activation (ligand-dependent or independent) of

members of the HER family

• RAS amplification

Amplification of MET contributes to acquired resistance to MET kinase inhibitors

GTL16 wt

Chromosome 7 centromere MET amplicon marker Wt PHA resistant (150 nM)

Giordano S. and coll. 2013

a-pMet a-Met

PHA 250 nM

Y1349 Y1356 P P P P P P S985 Y1003

D1228H (Kit) D1228N (Kit) Y1230C Y1230H M250T (Ret) M1131T V1188L L1196V V1220I

Some MET mutations confer resistance to MET inhibitors

Martin V, et al. Mol Oncol. 2014

SG16 Human gastro-esophageal Tumor SG16 P1 SG16 P2 Resistance

DN30 antibody treatment overcomes resistance to the MET kinase Inhibitor PHA-665752

PHA-665752 anti MET + DN30 antibody (complete remission) PHA-665752 anti MET (complete remission) Repeated PHA-665752 suboptimal concentrations

Carla Boccaccio: Cancer Stem Cell Laboratory Alberto Bardelli: Molecular Genetics Laboratory Silvia Benvenuti and Alessandra Gentile: Exploratory Research Group Maria Flavia Di Renzo: Laboratory of Cancer Genetics Pietro Gabriele: Dept of Radiotherapy Silvia Giordano: Laboratory of Molecular Biology Letizia Lanzetti Membrane Trafficking Laboratory Silvia Marsoni: Clinical Trial Unit Enzo Medico: Unit of Oncogenomics Luca Tamagnone Laboratory of Cancer cell Biology Livio Trusolino Unit of Translational Cancer Medicine Elisa Vigna Laboratory of Gene Therapy

The original data presented are synopsis taken from the work performed at the Candiolo Cancer Institute by :