Slide 1

vod do molekulrnej medicny MUDr. Ing. Bc. Peter Celec, PhD. Slide 2 2 Linus Carl Pauling (1901 1994) Nobel prize 1954 Chemistry 1962 Peace Slide 3 3 Life is a relationship between molecules, not a property of any one molecule. So is therefore disease, which endangers life. While there are molecular diseases, there are no diseased molecules. Linus C. Pauling, 1962 Slide 4 4 Basics Slide 5 5 Slide 6 Francis Crick 8.6.1916 28.7.2004 Slide 7 7 Slide 8 James Watson 6.4.1928 Slide 9 9 Slide 10 10 Slide 11 11 Molecular Medicine Based on molecular knowledge Based on XNA Proteomics Genomics Molecular interactions Regulations Signalling pathways Gene therapy Slide 12 12 What we will NOT talk about... XNA structure Monogenic disorders Immunology Carcinogenesis Slide 13 13 Genome ( DNA ) Transcriptome ( RNA ) Proteome ( protein ) Metabolome ( pathways ) Genome Slide 14 14 Slide 15 15 Information processing Communication Intercellular Intracellular Slide 16 16 Slide 17 17 Signalling pathways Slide 18 18 Ligands Hydrophilic Small molecules Larges proteins Hydrophobic Steroids Thyroxine Retinoids Slide 19 19 Receptors G-protein coupled Tyrosine kinases Ion channels Intracellular Slide 20 20 Signalling pathways Slide 21 21 Phosphate Slide 22 22 G-protein coupled Slide 23 23 G-protein coupled Animation Slide 24 24 Second messangers cAMP cGMP IP 3 DAG Ca ++ Slide 25 25 Gene The gene is by far the most sophisticated program around. Bill Gates Slide 26 26 Gene expression regulation Slide 27 27 Slide 28 28 Jak Janus (Roman god of gates and doors) Kinase Jak Just Another Kinase STAT Signal Transducers and Activators of Transcription APS Adaptor protein with Pleckstrin homology and Src homology 2 domains C-Cbl - Casitas B-lineage Lymphoma EPO Erythropoietin Slide 29 29 Tyk2 Tyrosine kinase 2 mTOR mammalian Target Of Rapamycin MAPK MAP (Microtubule Associated Protein) Kinase MAPK MAP (Mitogen Activated Protein) Kinase ISRE Interferon-Stimulated Response Element GAS IFN-Gamma-Activation Site Slide 30 30 GR Glucocorticoid Receptor FKBP FK506 (tacrolimus)-Binding Protein HSP Heat Shock Protein GRE Glucocorticoid Response Element Slide 31 31 c-Fos FBJ (Finkel MP, Biskis BO, Jinkins PB) OsteoSarcoma virus AP-1 Activator Protein 1 TGACTCA Slide 32 32 FAP FAP c-Fos AP-1 binding site IP-1 Inhibitory Protein 1 Slide 33 33 NFB Slide 34 34 FasL Fas Ligand TRAF TNF Receptor Associated Factor NIK NFB Inducing Kinase CBP CREB Binding Protein NFkB Slide 35 35 MAP kinases GEF Guanine Nucleotide Exchange Factor Slide 36 36 MAP kinases Animation Slide 37 37...but... Slide 38 38 Signalling pathways Slide 39 39 Responses??? Slide 40 40 Slide 41 41 Slide 42 42 Apoptosis Slide 43 43 Apoptosis Videos Slide 44 44 Slide 45 45 Slide 46 46 Autophagy MacroautophagyMicroautophagy Slide 47 47 Slide 48 48 Slide 49 49 Oxygen sensing Slide 50 50 HIF Hypoxia Inducible Factor IGFR Insulin like Growth Factor Receptor EGFR Epidermal Growth Factor Receptor HER2 Human Epidermal growth factor Receptor 2 PI3K PhosphatidylInositol-3-Kinase AKT serine/threonine protein kinase B FRAP FKBP rapamycin associated protein PH Prolin Hydroxylases HDAC histone deacetylase VHL von Hippel-Lindau tumour suppressor protein FIH factor inhibiting HIF-1 B elongin B C elongin C C2 cullin 2 HRE hypoxia response element GLUT glucose transporter ARNT Aryl hydrocarbon Receptor Nuclear Translocator Slide 51 51 Slide 52 52 Angiogenesis Slide 53 53 Slide 54 54 Slide 55 55 Aaron Ciechanover Avram Hershko Irwin Rose 1/3 of the prize Israel 1/3 of the prize Israel 1/3 of the prize USA Technion Israel Instutute of Technology Haifa, Israel Technion Israel Institute of Technology Haifa, Israel University of California Irvine, CA, USA b. 1947b. 1937 (Karcag, Hungary) b. 1926 The Nobel Prize in Chemistry 2004 "for the discovery of ubiquitin-mediated protein degradation" Slide 56 56 Ubiquitin Slide 57 57 Ub E1 Ub E2 Ub E3 Ub 26S proteasome degradation Target Ub Ub Ub Ubiquitin pathway Ubi-activating EUbi-conjugating EUbi-ligase Slide 58 58 Ub ATP AMP + PPi E1 Ub E1 C E2 C Ub E3 C Ub Protein K Ub E3 Protein K K E2 Ub E3 Ub Protein K E3 E2 C Ub Protein Ub AMP + PPi Protein K K K K K K K K Proteosynthesis Antigen presentation H A B C1 C2 D E Ub Protein Ub ATP Protein Ub Protein Ub I F G J Slide 59 59 Molecular diagnostics Slide 60 60 Kary Banks Mullis (1944 ) Nobel prize 1993 Chemistry Slide 61 61 PCR Slide 62 62 Slide 63 63 PCR Expensive? Slide 64 64 Slide 65 65 PCR Analyzed by Electrophoresis Slide 66 66 Real-time PCR Quantitative PCR Fluorescent detection of PCR products in real time Slide 67 67 RFLP Slide 68 68 RFLP & VNTR Animation Slide 69 69 Blotting Western Southern Northern Slide 70 70 Western Blot Slide 71 71 Western Blot Animation Slide 72 72 Southern Blot Slide 73 73 Slide 74 74 Southern Blot Animation Slide 75 75 RT PCR Animation Slide 76 76 FISH Procedure Slide 77 77 FISH Slide 78 78 Frederick Sanger (1918 ) Nobel prize 1958 Chemistry 1980 Chemistry Slide 79 79 Sequencing Animation Slide 80 80 Microarray Slide 81 2006 stav patologickej fyziolgie MUDr. Ing. Peter Celec, PhD. Gnov terapia Slide 82 DefinciaPrincpyRozdeleniaVektoryKlinikaProblmy Slide 83 Defincia Vetky prstupy na lieenie chorb, ktor vyuvaj nukleov kyseliny a gnov ininierstvo Slide 84 Princpy gnovej terapie Nhrada gnu Zabitie cieovej bunky Oprava mutcie Cielen inhibcia gnovej expresie Slide 85 Princpy gnovej terapie Nhrada gnu Slide 86 Princpy gnovej terapie Zabitie cieovej bunky Slide 87 Princpy gnovej terapie Zabitie cieovej bunky Slide 88 Princpy gnovej terapie Zabitie cieovej bunky indukciou imunitnho systmu Slide 89 Princpy gnovej terapie Oprava mutcie Slide 90 Princpy gnovej terapie Cielen inhibcia gnovej expresie Slide 91 Princpy gnovej terapie RNA interferencia Slide 92 Rzne delenia gnovej terapie Poda funknej nukleovej kyseliny DNA based RNA based Slide 93 Rzne delenia gnovej terapie Poda typu ovplyvnench buniek Somatick Zrodon Slide 94 Rzne delenia gnovej terapie Slide 95 Poda prstupu In vivo Ex vivo Slide 96 Rzne delenia gnovej terapie Poda prstupu Slide 97 Vektory VrusovNevrusovBakterilne Slide 98 Slide 99 Vrusov vektory RetrovrusAdenovrus Adeno-associovan vrus HerpesvrusAlfavrus... Slide 100 Slide 101 Nevrusov vektory Nah alebo obalen DNA Rzne fyzikochemick transportn systmy Slide 102 Nevrusov vektory PlazmidyPCR produkty Injekcia nahej DNA Elektroporcia Gene gun Lipoplexy Polyplexy Oligodeoxynukleotidy Slide 103 Bakterilne vektory Baktofekcia Listeria Escherichia Salmonella Alternatvna gnov terapia Slide 104 Baktofekcia Slide 105 Alternatvna gnov terapia Slide 106 Slide 107 Klinika 1990 prv klinick tdia Severe combined immunodeficiency (SCID) Retrovrusov vektor Integrcia do chromozmu Ex vivo Slide 108 Klinika Slide 109 Slide 110 Slide 111 Slide 112 Slide 113 Slide 114 Slide 115 Slide 116 Slide 117 Slide 118 Slide 119 Slide 120 Problmy GT Etick Prenos gnu innos pecificita Bezpenos Regulcia expresie Slide 121 Slide 122 September 11, 2001 February 15-16, 2001 Slide 123 Slide 124