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

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

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 Oncogene expedience (DNA) (mRNA) MET wt MET genetic alteration gene overexpression FREQUENT RARE Transcriptional induction : Chromosom. rearr . -hypoxia (TPR-MET, e.g. gastric K) -ionizing radiation Gene amplification (e.g. gastric and esoph. K; Gefitinib-resistant NSCL) Point mutations (e.g. Papillary kidney K; “cancer of unknown primary site”)

‘Oncogene addiction ’: • 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 ’.

‘ Primary oncogene addiction’: the response matches exactly Met amplification MET Cell Line copy N ° 197 cancer cell EBC-1 5.8 lines tested MKN-45 6 < 3 % ADDICTION GTL-16 6.1 HS746T 6.3 SNU5 5.6 NCI-H1993 5.2 J&J 605 Specific MET kinase inhibitor

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

The ‘ precision medicine’ approach (Liquid Biopsy followed by next gen. sequencing) Patient DM – Chromosome 17 Plasma MET Tumor MET (A.Bardelli et al . 2013)

SPEC-CT scan Imaging the MET oncogene amplification by 111 In DTPA-DN30 antibody Ovary Ca MET wt Lung Ca MET ampl. 12x 2h post injection 2h post injection

Response to specific inhibitors by a ‘xenopatient’ bearing a MET amplified colorectal Ca. 3000 Xenopatient M162 Vehicle 2500 Cetuximab Tumor Volume (mm 3 ) 2000 Crizotinib Cetuximab+Crizotinib 1500 JNJ-38877605 1000 Cetuximab+JNJ- 38877605 500 0 -40 -30 -20 -10 0 10 20 30 40 50 Days L.Trusolino et al, 2014

MET amplification is associated with secondary addiction in anti-EGFR resistant patients A MET MET CEP7 CEP7 Met IHC 60X 40X 60X 40X Patient #1: before Pmab Patient #1: after Pmab B MET MET CEP7 CEP7 Met IHC 40X 40X 60X 60X Patient #2: before Pmab Patient #2: after Pmab C MET MET CEP7 CEP7 Met IHC 40X 60X 40X 60X Patient #3: before Cmab Patient #3: after Cmab 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

‘Oncogene expedience ’: • 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 ’).

Ionizing radiations activate the MET oncogene Ionizing radiations MET overexpression IKKa-b Nemo P P ROS P P p50 P RelA I  B NF  B ATM DNA damage Radioresistance (...) MET 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 m g/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

Oncogene ‘ Inherence ’: • 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’) .

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 O OH n • 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”)

Non-conventional reponse to MV-DN30 Monoclonal Antibody Ligand Ligand neutralization Inactive p125 p125 Receptor heterodimer Decoy MV-DN30 Inhibitory effects Shedding Γ - secretase Adam 10 Proteasome degradation p55 Met p50 p175 Reviewed by: Vigna E. and Comoglio P.M., Oncogene.34:1883-89; (2015)

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

An orthotopic mouse model of human GBM LV- MvDN-30 vector

Gene therapy with LV( Tet )-DN30 FAb (U87 Glioblastoma ) 100 90 % tumor-free animals . 80 70 60 no dox ( Fab-) 50 dox + (Fab+) 40 30 20 10 0 0 10 20 30 40 50 60 70 80 90 100 110 Time (days) 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 PHA resistant (150 nM) Wt Chromosome 7 centromere GTL16 MET amplicon marker wt Giordano S. and coll. 2013

Some MET mutations confer resistance to MET inhibitors PHA 250 nM M1131T P S 985 Y 1003 V1188L P D1228H ( Kit ) a-pMet L1196V D1228N ( Kit ) V1220I P Y1230C P Y1230H M250T ( Ret ) a-Met P Y 1349 Y 1356 P Martin V, et al. Mol Oncol. 2014

DN30 antibody treatment overcomes resistance to the MET kinase Inhibitor PHA-665752 Repeated PHA-665752 suboptimal concentrations SG16 P2 SG16 SG16 P1 PHA-665752 anti MET Resistance Human gastro-esophageal + Tumor DN30 antibody (complete remission) PHA-665752 anti MET (complete remission)

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