Manifestation of Novel Social Challenges of the European Union in the Teaching Material of Medical Biotechnology Master’s Programmes at the University of Pécs and at the University of Debrecen Identification number: TÁMOP /1/A

TYROSINE KINASE- LINKED RECEPTORS PART 2 GROWTH FACTORS Tímea Berki and Ferenc Boldizsár Signal transduction Manifestation of Novel Social Challenges of the European Union in the Teaching Material of Medical Biotechnology Master’s Programmes at the University of Pécs and at the University of Debrecen Identification number: TÁMOP /1/A

TÁMOP /1/A Growth factors (GFs) Small molecular weight soluble mediators They control: 1 1Proliferation 2 2Survival 3 3Metabolism 4 4Tissue differentiation Important implication in tumors Cytokines – growth factors

TÁMOP /1/A Growth factors (GFs) – History How to propagate cells under in vitro conditions (turning of the 19 th -20 th century)? Rous: experiments with chicken tumor (sarcoma) cells (RSV) Carrel: In simple buffered salt solution the cells did not proliferate – trials with diluted plasma/serum Temin and Dulbecco: precise requirements for tissue culturing: –Reduced serum need of tumor cells – enhanced capacity of tumor cells to respond to proliferation signals –Serum supported cell growth rather than plasma – PDGF Rita Levi-Montalcini, Stanley Cohen – NGF and EGF Transplantation of an actively growing mouse tumor into chicken induced the great amplification of nerve fibres into the tumor mass

TÁMOP /1/A Growth factors FactorPrincipal sourcePrimal activityComments PDGF Platelets, endothelial cells, placenta Promotes proliferation of connective tissue, glial and smooth muscle cells Two differentprotein chains from 3 distinct dimer forms; AA, AB and BB EGF Submaxillary gland, Brunners gland Promotes proliferation of mesenchymal, glial and epithelial cells TGF-αCommon in transformed cells May be important for normal wound healing Related to EGF FGF Wide range of cells; protein is associated with the ECM Promotes proliferation of many cells; inhibits some stem cells; induces mesoderm to form in early embrios At least 18 family members, 5 distinct receptors NGF Mast cells, eosinophils, bone marrow stromal cells, keratinocytes Promotes neurite outgrowth and neural cell survival Member of a family of proteins termed neutrophins that promote proliferation and survival of neurons; neutrophin receptors are a class of related proteins first identified as proto-oncogenes? TrkA („trackA”), TrkB, TrkC ErythropoietinKidneyPromotes proliferation of erythrocytes TGF-β Activated Th1 cells (T-helper) and natural killer (NK) cells Anti-inflammatory (suppresses cytokine production and class II MHC expression), promotes wound healing, inhibits macrophage and lymphocyte proliferation At least 100 different family members IGF-1Primarily liverPromotes proliferation of many cell types Related to IGF-2 and proinsulin, also called somatomedin C IGF-2Variety of cells Promotes proliferation of many cell types primarily of fetal origin Related to IGF-1 and proinsulin

TÁMOP /1/A Receptors with TK activity Receptor Tyrosine Kinases (RTK) e.g. PDGF, insulin, EGF, VEGF and FGF receptors Tyrosine-Kinase Associated Receptors Receptors that associate with proteins that have tyrosine kinase activity (Cytokine Receptors) Receptor Tyrosine Phosphatases e.g. CD45 protein of T cells and macrophages

TÁMOP /1/A “Complete” and “incomplete” receptor tyrosine kinase “Complete” receptor tyrosine kinase “Incomplete” receptor Cytoplasmic non-receptor tyrosine kinase Adaptor SignalSignal

TÁMOP /1/A Receptor tyrosine kinase (RTK) families 90 unique Tyr kinases in the human genome, 58 are RTKs Growth factor, cytokine and hormone receptors Classes: I EGFR family (ErbB) XLTK family II Insulin rec. family XITIE family IIIPDGF familyXIIROR family IVFGF familyXIIIDDR family VVEGF familyXIVRET family VI HGF family (c- Met) XVKLG family VIITrk familyXVIRYK family VIIIEph familyXVIIMuSK family IXAXL family

TÁMOP /1/A Kinase-phosphatase balance Phosphorylase kinase (ser/thr kinase) PP1c (ser/thr phosphatase) Phosphorylase b Phosphorylase a P P InactiveActive P ATPADP CD45 (tyr phosphatase) Csk (tyr kinase) ADPATP Inactive p56 Lck P Y505 Y394 Primed p56 Lck Y505 Y394 Active p56 Lck P Y394 P

TÁMOP /1/A Fibronectin III Leucine-rich Cysteine-rich Acid-box Kinase IG-like VEGFR1 VEGFR2 VEGFR3 PDGFR  PDGFR β CSF1R Kit Kit2 RykTorsoEGFR ErbB2 ErbB3 ErbB4 Met Ron Sea TrkA TrkB TrkC INSR IGF1R IRR Axl Mer Sky Eph Eck Eek Erk Elk Ehk1 Ehk2 Sek Hek Hek11 Cek-9 Myk-1 Myk-2 RosFGFR1 FGFR2 FGFR3 FGFR4 Tie Tie2 DDRRetTorpedoRor1 Ror2 Ltk Alk EGF-like Cadherin Factor VIII-like Glicyne-richKringle C1r-like Growth factor receptors

TÁMOP /1/A Receptor-like PTPs (21) CD45 (RC) R1/R6 PTP  (RM) PTP  (RK) PTP  (RT) PTP (RU) R2B LAR (RF) PYP  (RS) PTP  (RD) R2A PTP  (RA) PYP  (RE) R4 PTP  (RG) PYP  (RZ1) R5 PTP β (RB) DEP1 (RJ) SAP1 (RH) GLEPP1 (RO) PTPS31 (RP) R3 PCPTP1  (RR) STEP (N5) R7 IA2 (RN) IA2 β (RN2) R8 RGDS motif Proline-richMAM domain SEC14 domainFibronectin III FERM domain IG-like BRO-1 homologyGlycosylated PDZ domainCadherine-like Histidine domainCarbonic anhydrase-like Kinase-interacting domainPTP domain Src homology 2 PTP pseudo-phosphatase domain

TÁMOP /1/A Nontransmembrane PTPs (17) HDPTP (N23) NT8 MEG2 (N9)NT3 HePTP (N7) PTPH1 (N3) MEG1 (N14)NT5 SHP1 (N6) SHP2 (N11)NT2 PTPBAS (N13) NT7 PTPD1 (N21) PTPD2 β (142) NT6 PTP1B (N1) TCPTP β (N2)NT1 BDP1 (N18) PTP-PEST (N12) LYP (N220)NT4 PTPTyP (N20) NT9 RGDS motif Proline-richMAM domain SEC14 domainFibronectin III FERM domain IG-like BRO-1 homologyGlycosylated PDZ domainCadherine-like Histidine domainCarbonic anhydrase-like Kinase-interacting domainPTP domain Src homology 2 PTP pseudo-phosphatase domain

TÁMOP /1/A Growth factor receptors and tyrosine phosphorylation p120 Ras-Gap p120 Ras-Gap PLC  Phosphotyrosine Y559 Y581 Y716 Y741 Y751 Y771 Y1009 Y1021Phosphotyrosine Y992 Y1045 Y1068 Y1086 Y1148 Y1173 PDGFR P P P P P P P P P P P P P P EGFR P P P P P P P P P P

TÁMOP /1/A Receptor tyrosine kinase (RTK) signalingTranscription RTK Ligand P P P P P P P P Dimerization Src PKC PLC STAT JAK Akt

TÁMOP /1/A I GF receptor signaling pathways I Targets PIP 2 PIP 3 Targets Akt PDK1 PIP 3 SOS Ras Targets Erk Targets GRB2 Shp2 GRB2 PI3K RTK Ligand PP P P P P P P P Plasma membrane P P P P P P P P

TÁMOP /1/A II GF receptor signaling pathways IIRTK Ligand PIP 2 Cbl P PLC C2PHSH3SH2 Plasma membrane Cytoplasm P P P P P P P P

TÁMOP /1/A Overview of EGF signaling

TÁMOP /1/A General characteristics of GF signaling Diverse input signals (Multiple RTKs) Diverse input signals (Multiple RTKs) Conserved core processes Conserved core processes Diverse ouput events (transcriptional responses, cytokeletal changes, etc) Diverse ouput events (transcriptional responses, cytokeletal changes, etc) System control Input layer Output layer

TÁMOP /1/A Different GF receptors use the same signaling pathways PDGF-C Cell survival Proliferation Apoptosis resistance Metastasis Angiogenesis P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P EGFRHer2Her3Her4VEGFR1VEGFR2VEGFR3PDGFR-aPDGFR-bc-kit EGF TGF  β-cellulin Amphiregulin HB-EGF No specific ligands Heregulins β-cellulin NRG2 NRG3 VEGF-BVEGF-AVEGF-CVEGF-DPDGF-A PDGF-B PDGF-D SCF SOS GRB2 Ras Rac Rho CDC42 MEK1/2 Erk Raf ERK pathway MKK4/7 JNK MEKK JNK pathway MKK3/6 p38 Tak p38 pathway Akt mTor PI3K Everolimus Imatinib Trastuzumab Leflunomide Lapatinib Gefitinib Erlotinib Panitumumab Sorafenib Cetuximab Bevacizumab Vandetanib Sunitinib Enzastaurin Pazopanib Motesanib Midostaurin Temsirolimus Sirolimus P P P P P P P P

TÁMOP /1/A Natriuretic peptide signaling ↑NP degradation ↓cAMP? ↑ IP3? ↑ Vasorelaxation ↑ Diuresis, natriuresis ↓ Renin, aldosterone ↓ Cell proliferation ↓ Cardiac fibrosis ↑ Vasorelaxation ↓ Cell proliferation ↑ Long bone gowth Kinase homology domain Plasma membrane Ligand binding domain Receptor Hinge region Guanylyl cyclase domain Physiologic response Natriuretic peptide NPR-CNPR-A (GC-A) NPR-B (GC-B) ANPBNPCNPcGMPGTPcGMPGTP P P P P P P P P P P P P P P P P P P P P P P Natriuretic peptide Hormone bound Active Desensitised Kinase Phosphatase ATP cGMPGTP PP P PP PP Basal ATP