1. dia

Manifestation of Novel Social Challenges of the European Unionin the Teaching Material ofMedical Biotechnology Masters Programmesat the University of Pcs and at the University of DebrecenIdentification number: TMOP-4.1.2-08/1/A-2009-00111Cell Cycle and CAncer

Zoltan BalajthyMolecular Therapies- Lecture 12Manifestation of Novel Social Challenges of the European Unionin the Teaching Material ofMedical Biotechnology Masters Programmesat the University of Pcs and at the University of DebrecenIdentification number: TMOP-4.1.2-08/1/A-2009-00112Learning objectives of chapter 12 and 13 . The purpose of this chapter is to describe the processes and regulations of both cell cycle and cell death, explain the unregulated cell division, and to point out the therapeutic intervention in cancer at molecular levels. Topics in chapter 12.

12.1. Interpretation of cell cycle Constitutive and Inducible Cell Cycle Kinase Inhibitors12.2. Mitogen-activated Protein (MAP) Kinase Cascade Transcriptional Events in G1 Phase of Cell-cycle Mechanisms of Gene-suppression by the Retinoblastoma Protein 12.3. Biochemical Events of Cell-cycle in M Phase 12.4. Cancer Causing Genes in Mitotic Signal Pathway Proto-oncogenes and OncogenesTMOP-4.1.2-08/1/A-2009-001112.5. Different Families of Receptor Tyrosine Kinases (RTK) Proto-oncogenes are normal genes that can become oncogenes 12.6. Therapeutic TargetsMonoclonal antibodies and specific inhibitorsTMOP-4.1.2-08/1/A-2009-00113

CheckpointscontrollThe conventional cell cycle is modified to indicate :- G1 activities, those preparatory for S phase, may begin during the previous cycle, concurrently with G2 and mitotic events. early preparation for mitosis (G2) may overlap with S - exit from G1 into the G0 quiescent state and reentry into the cycle

The critical points: C (indicating competence), V (end of entry)R (restriction point, end of progression)12.1. The Functional Cell Cycle TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 1. The Functional Cell Cycle 4Cyclin-CDK Regulators of Cell-Cycle

TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 2. Cyclin-CDK Regulators of Cell-Cycle

5

cell divison cycle: CDC,cyclin-dependent protein kinases: CDKcyclin-dependent protein kinases inhibitor: CDI, CDK1CKIsDNA damade,starvation orfactor inducibleDNA-damageinducibleConstitutiveCKIConstitutiveCKIAPCConstitutive and Inducible Cell Cycle Kinase Inhibitors, and arecheck pointsat the s, gm,and m stable statesTMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 3. Constitutive and Inducible Cell Cycle Kinase Inhibitors

612.1. Mitogen-activated Protein (MAP) Kinase Cascade

TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 4. Mitogen-activated Protein (MAP) Kinase Cascade

7

CDK inhibitorsCDK inhibitorsCDK inhibitorsDihydrofolate reductaseThymidine kinase Thymidilate synthase DNA polymeraseE2F: transcription factor E2F1 EGF: epidermal growth factorCDK: cyclin-dependent protein kinaseRb: retinoblastoma proteinD1, A, E: Cyclin D1, A s EDNS replication machineryStart of S phasepozitv ersts transzkripcilelltsTranscriptional Events in G1 Phase of Cell-cycle DNS replication machineryTMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 5. Transcriptional Events in G1 Phase of Cell-cycle

8

Mechanisms of Gene-suppression by the Retinoblastoma Protein TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 6. Mechanisms of Gene-suppression by the Retinoblastoma Protein

911.3. Biochemical Events of Cell-cycle in M Phase

TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 7. Biochemical Events of Cell-cycle in M Phase

1012.4. Cancer Causing Genes in Mitotic Signal Pathway

TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 8. Cancer Causing Genes in Mitotic Signal Pathway

11Proto-oncogenes and Oncogenes

TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 9. Proto-oncogenes and Oncogenes

12Possible Biochemical Mechanisms of Protooncogene - oncogene Conversion

- promoter insertion- enhancer insertion- chromosomal translocation- gene amplification- single point mutation or deletionOut the 5 mechanisms described above, the first 4 involve an increase in amount of the product of an oncogene due to increased transcription but no alteration of the structure of the product of the oncogene. Thus it appears that increased amounts of the product of an oncogene may be sufficient to push a cell becoming malignant. The fifth mechanism, single point mutation, involves a change in the structure of the product of the oncogene but not necessarily any change in its amount. This implies that the presence of a structurally abnormal key regulatory protein in a cell may be sufficient to tip the scale toward malignancy.TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 10. Possible Biochemical Mechanisms of Protooncogene - oncogene Conversion

13EGFR(HER 2,3,4TGF-aEGFIGF-1RPPPPIGF-1 IGF-2PDGFRPDGFVEGF-A VEGF-BVEGFRHER2NoneEGFR/HER2PPPPReceptor Heterodimerization and Activation12.5. Different Families of Receptor Tyrosine Kinases (RTK) Recognize a Diverse Set of Different LigandsTransmembraneLipophilic SegmentIntracellular ProteinTyrosine Kinase DomainExtracellular Binding DomainTMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 11. Different Families of RTKs Recognize a Diverse Set of Different Ligands

14EGFR-s(HER3,4)TGF-aEGFEGFRKinase CatalyticSiteCytosolic DomeinExtracellularDomeinReceptor HomodimerizationLigand-binding SiteLigandPPPPATPADPLigand Binding Activates RTKs by DimerizationTMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 12. Ligand Binding Activates RTKs by Dimerization

15Proto-oncogenes are Normal Genes That Can Become Oncogenes

TMOP-4.1.2-08/1/A-2009-0011Figure 13. Proto-oncogenes are Normal Genes That Can Become Oncogenes

16mAbsLXXSignal transductiomXLXXSignal transductiomXLTyrosine kinase inhibitors (TKI)TKITKILLCell deathToxin-induced cell deathLLRNA interference ProteinsynthesisXNeu, EGFR-targeting MethodsTMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 14. Neu, EGFR-targeting Methods

17

Expression of HER2 Receptor on the Surface of Normal and Tumor CellsHerceptin Binds to the HER2 Receptor on the Surface of Tumor Cells Normal expression ofHER2 receptorIncreased expression of HER2 receptorHerceptin attaches itself to HER2 receptor Herceptin flags the cancer cells for destruction by the immune systemTMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 15. Expression of HER2 Receptor on the Surface of Normal and Tumor Cells Herceptin Binds to the HER2 Receptor on the Surface of Tumor Cells

181, blocking signalization on cell surface receptor

2, inhibition of TK activity of TKR

3, inhibition of protein kinases phosphorilation cascade

4, activation of growths-inhibiting pathway (Rb) or restoration of p53 function

5, inhibition of angiogenesis6, inhibition of methastasis In addition to the classical form of cancer treatment: like surgery, chemotherapy and radiotherapy, there are new therapeutic targets at molecular level against cancer cells.1TKI23

4TKI12.6. Therapeutic TargetsTMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 16. Therapeutic Targets

19Blocking of Oncoproteins of EGFR and Mitogen-activated Protein Kinase Signalization via Monoclonal abs. and Specific Inhibitors

TMOP-4.1.2-08/1/A-2009-0011TMOP-4.1.2-08/1/A-2009-0011Figure 17. Blocking of Oncoproteins of EGFR and Mitogen-activated Protein Kinase Signalization via Monoclonal abs. and Specific Inhibitors

20