C-MET

1. For more presentations and information visit http://www.pharmaxchange.info C-MET By Tanay Surkund Roll no : 60 Guide: Mrs. Poonam Advani

2. For more presentations and information visit http://www.pharmaxchange.info C-MET -The Receptor Tyrosine Kinase • Proto-oncogene coding for the C-MET protein • Protein essential for organ protection and wound healing. • Activation of MET gives rise to the invasive growth programme. • Abnormal MET activation causes cancer growth in areas such as the kidneys ,lungs ,stomach ,liver, and brain. • Found in the epithelial tissues of all the above mentioned organs. • HGF is the only known ligand

3. For more presentations and information visit http://www.pharmaxchange.info C-MET protein • Disulphide linked heterodimer, which results from cleavage of a precursor at a furin site. • It is a trans-membrane protein and consists of : 1) Extracellular region: which consists of the alpha and beta chains, the sema domain, the MRS and the immunoglobulin structures. 2) Intracellular region: which consists of the juxtramembrane segment, the tyrosine kinase domain and the C-terminal region which comprises 2 crucial tyrosines. • Signalling activity on formation of a dimer.

4. For more presentations and information visit http://www.pharmaxchange.info C-MET Protein structureExtracellular region • Extracellular region • It consists of an alpha and a beta chain. The latter is much longer and spans the entire ectodomain,transmembrane helix and cytoplasmic portion.The alpha region is joined to the beta chain by a disulphide linkage. • The alpha portion and the first 212 residues of the beta chain are sufficient for ligand binding with HGF/SF. • This region is homologous to the sema domain of the semaphorin axon guidance system and folds into a beta propeller structure. • The rest of the ectodomain consists of a cysteine rich sequence, Met related sequence(MRS), followed by glycine-proline rich (G-P) repeats and four unusual immunoglobluin structures.

5. For more presentations and information visit http://www.pharmaxchange.info C-MET Protein Structure • Intracellular Region It consists of 3 regions: • A Juxtramembrane segment consisting of: • A serine residue which on phosphorylation by protein kinase or calcium calmodulin dependant kinases,downregulates the receptor kinase activity. • A Tyrosine(Tyr 1003) that binds to ubiquitin ligase Cbl, which is responsible for Met polyubiquination,endocytosis and degradation. • The Tyrosine kinase domain, bearing tyrosine 1234 and tyrosine 1235 which cause transphopshorylation of other tyrosine residues on the other MET molecule of the dimer. • The C-terminal region comprises 2 crucial tyrosines inserted in a degenerative motif that represents a multisubstrate docking site capable of recruiting several downstream adaptors containing Src-homology 2(SH2) domain. • Met receptor as most receptor tyrosine kinases use different tyrosines to bind to specific signalling molecules.The 2 docking sites are essential for signal transduction both invitro and invivo.

6. For more presentations and information visit http://www.pharmaxchange.info MET Signaling PathwayI. Components • MET receptor bears 2 unique bidendate docking sites which comprimise tyrosine residues Tyrosine 1349(Y1349) and Tyrosine 1356.For Steric reasons,two substrates cannot bind simultaneously to the bidendate site of one Met molecule. • When transphosphorylated by the Kinase domains, they bind to Gab-1, Grb-2, phosphatidinylinositol-3-kinase(PI3K) and others.Most of these components contain Src-homology-2(SH2) domain binding sites which mediate the reactions. • SH2 is the largest class of pTyr- recognition domains, and typicallty bind a phosphorylated tyrosine residue in the context of a longer peptide motif with a target protein.

7. For more presentations and information visit http://www.pharmaxchange.info MET Signaling PathwayII. Transducer binding with Gab-1 • Versatile, Scaffolding adaptor protein • Most crucial binding substrate. • Phosphorylation causes binding to the Shp2,PI3K,phospholipase C and crk in which they act downstream and the Ras and Raf upstream.(MAPK componenets) • Shp2 upregulates the MAPK and Erk pathway.

8. For more presentations and information visit http://www.pharmaxchange.info MET signalling pathwayIII. Pathway activation and functions. • Activation of the MAPK/ERK pathway: • Cell proliferation through activation of pBR and Cdk6. Both of which are important in cell cycle progression. • Activation of Fibronectin( Wound healing) • Activation of Integrins- leads to cell adhension and is regulated by FAKs.(Cell adhesion and migration) • Activation of the Pi3K/Akt pathway: • Anti-apoptosis by deactivation of Bad or through a Caspase-9 cleavage.

9. For more presentations and information visit http://www.pharmaxchange.info MET signalling pathwayIII. Pathway activation and functions. • Activation of the Ras pathway which consecutively activate the RHO, RAC1 and CDC42 pathway: • Cytoskeletal rearrangement via the arp2/3 complexes • Cell mobility via formation of lamellopodia by actin rearrangement • Cadherin rearrangement

10. For more presentations and information visit http://www.pharmaxchange.info Dysregulation of MET receptor tyrosine kinase in the formation of invasive growth tumours. • Increased MET expression has been found in papillary carcinomas of the thyroid gland, in carcinomas of colon, pancreas, and ovary, in osteogenic sarcomas, and in other types of cancer. Point mutations in MET have been identified in hereditary and sporadic papillary renal carcinomas , hepatocellular and gastric carcinomas and head and neck squamous carcinomas. • There are 3 mechanisms by which tumours are formed due to the met receptor. • Ligand dependant mechanisms of MET activation • Ligand independant mechanisms of MET activation • MET transactivation by other membrane receptors

11. For more presentations and information visit http://www.pharmaxchange.info Dysregulation of MET receptor tyrosine kinase in the formation of invasive growth tumours. • Ligand Dependant mechanisms of met activation • Tumor cells express both MET and HGF setting an autocrine loop in which, secreted HGF binds to the MET receptor and causes the activation of its downstream signaling pathways thus enhancing tumor growth and invasive behavior. • Such loops have been detected in gliomas, osteosarcomas, and mammary, prostate, breast, lung and other carcinomas. • This mechanism of tumor formation is not only dependant on MET and HGF but also on the enzymes responsible for the proteolytic activation of HGF from the single chain precursor molecule pro-HGF.A number of serine proteases, including urokinase type plasminogen activator and coagulation factor XII have been detected in some tumors.

12. For more presentations and information visit http://www.pharmaxchange.info Dysregulation of MET receptor tyrosine kinase in the formation of invasive growth tumours. • Ligand Independent mechanisms of MET activation • MET overexpression • Gene Rearrangement (TPR-MET) • Absence of negative regulators • Missense Mutations • MET transactivation via other membrane receptors • CD44 cell surface receptors present for hyaluronic acid • Integrins • Ron receptors

13. For more presentations and information visit http://www.pharmaxchange.info CANCER THERAPIESTARGETTING HGF/MET • MET kinase inhibitors • K252a(Fermentek) • ARQ-197(ArQule) • XL880(Exlixis) • PHA-6657572(Pfizer) • HGF inhibitors • NK4 • Neutralizing anti-HGF antibodies • Uncleavable HGF • DECOY MET • CGEN241

14. For more presentations and information visit http://www.pharmaxchange.info CANCER THERAPIESTARGETTING HGF/MET • IMMUNOTHERAPY TARGETTING MET • PASSIVE IMMUNOTHERAPY • DN30 • OA-5D5 • ACTIVE IMMUNOTHERAPY • INTERFERONS • INTERLEUKINS

15. For more presentations and information visit http://www.pharmaxchange.info References • www.nature.com • www.PubMED.com- journals • MET,METASTASIS,MOTILITY-Carmen Birchmeier*,Walter Birchmeier‡, Ermanno Gherardi§ and George F.Vande Woude. 2003 Nature Publishing Group • Hepatocyte Growth Factor Establishes Autocrine and Paracrine Feedback Loops for the Protection of SkinCells after UV Irradiation. Journal of Investigative Dermatology (2007), Volume 127 • Potent and selective inhibitors of the Met [hepatocyte growth actor/scatter factor (HGF/SF) receptor] tyrosine kinase block HGF/SF-induced tumor cell growth and invasion.LETTERS TO NATURE. • Anti-apoptotic signaling by hepatocyte growth factor by Met via the phosphatidylinositol 3-kinaseyAkt and mitogen-activated protein kinase pathways.PNAS • Transcriptional activation of the Hepatocyte Growth Factor receptor (c-met) gene by its ligand (Hepatocyte Growth Factor) is mediated through AP-1. Oncogene (2000) ã 2000 Macmillan Publishers Ltd. • www.pnas.org • Invasive growth: from development to metastasis. J. Clin. Invest. 109:857–862 (2002). DOI:10.1172/JCI200215392.

16. For more presentations and information visit http://www.pharmaxchange.info References • Dysregulation of Met receptor tyrosine kinase activity in invasive tumors. J. Clin. Invest. 109:863–867 (2002). DOI:10.1172/JCI200215418. • The Met tyrosine kinase receptor in development and cancer. Cancer Metastasis Rev (2008) 27:85–94 DOI 10.1007/s10555-007-9107-6. • "Severe diabetes, age-dependent loss of adipose tissue, and mild growth deficiency in mice lacking Akt2/PKB beta" Journal of Clinical Investigation Volume 112 • Phosphatidylinositol 3-kinase regulates the induction of long-term potentiation through extracellular signal-related kinase-independent mechanisms. The Journal of neuroscience : the official journal of the Society for Neuroscience . • GTP-binding proteins of the Rho/Rac family: regulation, effectors and functions in vivo.". Bioessays29 (4): 356–370.doi:10.1002/bies.20558. PMID 17373658.