Il progetto Loxina - Irene Predazzi PPT
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20 Ottobre 2009 Il web a supporto della ricerca biomedica Il progetto Loxina Irene M. Predazzi Sezione di Genetica Dip. Biopatologia e Diagnostica per Immagini ... complessità del problema... età rapporto geni/ambiente CARDIOPATIE Non ischemiche SECONDARIE PRIMARIE Ipertensione HCM Miocardite CMD Da agenti tossici ARVD Altro... RCM Ischemiche ACUTA CRONICA > 35-40 anni: 20-160/100.000 80% ≤ 35 anni: 1.3-8.5/100.000 10% Altro... DIFETTI DEI CANALI IONICI BRUGADA QT LUNGO QT CORTO TVP WPW Altro... COMPLEX DISEASE‘S HETEROGENEITY Genetic factors Gene+ Gene+ Interaction Gene - Interaction Gene Interaction - Interaction Gene + Interaction Complex phenotype Gene+ Interaction altro Smoke Stress Interaction Diet Environmental factors modificata da Monika Stoll, Ph.D, Physiological Genomics from Rats to Human Process Endothelial cell disfunction Endothelial cell activation Inflammation Proteolysis Apoptosis Target Flowmediated vasodilation Adhesion molecules Macrophages MMPs Cathepsins Approximate AHA lesion stage I II III Lipid-core and fibrouscap formation Lipid core Fibrous cap IV Angiogenesis Thrombosis αVβ3-Integrin Fibrin Platelets (αVβ3Integrin) Tissue factor V VI Watkins et al 2006 Lipid metabolism, inflammatory and thrombotic pathways involved in coronary artery CD14, TLR4, IL1, IL6, TNFα, PAPP-A, RANTES… Inflammation: Primary cytokines Putative innate immunity susceptibility OX40L co-stimulatory molecule Upregulation of cytokines and adhesionmolecules in endothelium Putative innate immunity susceptibility Leucocyte infiltration CRP release from liver FLAP eNOS, ACE, ATIR…. Pro-atherogenic lipoprotein and lipid changes Leukotrienes Hyperlipidaemia-dependent inflammation and macrophage entry Known monogenic and QTL influences High LDL Low HDL Prothrombotic state Atherosclerosis Recently implicated genes that are involved in lipid metabolism LDLR, LIPC, LIPG, MVK, LDLR, OLR1, LCAT, APOE, APOC1, APOC4, APOB, PCSK9, LPL, CETP, APOA5, USF1… Plaque rupture Known monogenic and QTL influences PAI-1, MTHFR, THBS4… Thrombosis Watkins et al 2006 LOX-1 (Lectin Like low-density lipoprotein receptor-1) Ohki et al 2005 Ohki et al 2005 OLR1 – INTRONIC LINKAGE DISEQUILIBRIUM BLOCK rs11053654 C/T rs2742116rs3912640 A/T A/T rs2742115 A/G rs12308385 A/G rs2742113 A/C rs2010655 rs11053648 A/G C/T rs3736235 A/G rs11053646 rs11611453 rs6488265 C/G A/G A/G K167N rs2634161 A/C rs12309394 C/T rs3736233 A/G rs2742118 A/C rs1050283 C/Trs12231585 rs3816844 C/T A/G rs1050755rs11609310 A/G A/G rs11053645 A/T 3’ UTR 5’ UTR ex1 rs2742111 A/T rs2634163 A/G int1 rs2634162 A/T ex2 int2 ex3 rs12316417 A/C rs3741860 C/G rs2742114 A/G/T rs2742117 C/G int3 ex4 int4 rs11611438 C/G rs2742109 A/C ex5 int5 rs3736232 C/G rs12822177 A/G rs11053649 G/T rs3736234 C/T rs2634156 C/T ex6 rs7964641 A/Trs11615002 rs1050286 A/G A/T rs12231578 A/G rs1050289 A/Grs3994135 A/G LOX-1 (Lectin Like low-density lipoprotein receptor-1) Citoplasmic tail Trans membrane region CTLD domain Neck 5’ CTGGTT 3’ LOX-1 (complete receptor) mRNA DNA Exon 1 Exon 2 Exon 4 Exon 3 Exon 5 A/G GCAAGC G/T Loxin (lacking ofG/C exon 5) Exon 6 G/T A/G C/T OLR1 (oxidised low density lipoprotein (lectin-like) receptor 1) Single Nucleotide Polymorphisms on OLR1 Mango et al 2005 MEASUREMENTS: ASSOCIATION ON INDEPENDENT SAMPLES Reference Phenotype rs3736235 Ancestral allele rs11053646 (C)N allele Mango et al. 2003 (n = 103 vs 150) Miocardial Infarction Protective Protective Tatsuguchi et al. 2003 (n = 102 vs 102) Miocardial Infarction --- Risk Ohmori et al. 2004 (n = 167 vs 419) Severity of CAD --- Protective Sentinelli et al. 2006 (n = 351, n = 251) Miocardial Infarction No association --- ADVANCE study* (Knowles et al, 2008, n = 1200 vs 1045) Miocardial Infarction Protective Protective ARIC study* (Knowles et al, Miocardial 2008, n = 1156 vs 8702) Infarction Risk No association Hou et al 2009 (n = 284 vs 280) --- Risk Hypertension Irene Marta Predazzi 24/05/07 OLR1 INTRONIC SNP GENOTYPING PILOT ANALYSIS ON ~ 300 samples VARIABLES’ AND SAMPLE CRITERIA DEFINITION NEW SAMPLES COLLECTION SNP GENOTYPING DATA ANALYSIS SAMPLE CRITERIA Cases Controls Inclusion ST segment elevation myocardial infarction (STEMI) Absence of atherosclerotic lesions confirmed by angiographic data Inclusion First event Positive for at list one risk factor for Coronary Artery Disease Inclusion 55 ≤ age ≥ 70 55 ≤ age ≥ 70 Inclusion Angiography data availability Angiography data availability Exclusion Familial hypercholesterolemia Familial hypercholesterolemia Exclusion Drug induced suspect --- Exclusion Dialitic treatment STEMI Dialitic treatment VARIABLE’S DEFINITION - Anagraphic - Risk Factors - Clinical - Previous Treatments OLR1 INTRONIC SNP GENOTYPING Osp. S. Raffaele Istituto Clinico Humanitas Ist. Monzino Osp. Brescia Un. degli Studi di Genova Osp. Sandro Pertini Un. La Sapienza Centro Interdipartimentale di Osp. S. Eugenio Biostatistica e Bioinformatica (CIBB) Un. Tor Vergata Università di Tor Vergata DATA ANALYSIS SAMPLE RECRUITMENT OBIETTIVI - Reclutamento di un campione rappresentativo della popolazione italiana per lo studio dell’Aterosclerosi - Messa a punto del kit - Definizione del rischio associato ai polimorfismi su OLR1 - Messa a punto di un sistema di trasmissione dati elettronico RINGRAZIAMENTI Prof. Giuseppe Novelli Prof.ssa Carla Rossi Dott.ssa Francesca Amati Prof.ssa Alessandra Nardi Dott.ssa Lucia Vecchione Prof. Gianpaolo Scalia Tomba Dott. Ruggiero Mango Dott.ssa Simona Iacobelli Prof. Francesco Romeo Dott. Saverio Muscoli Dott. Riccardo Morgagni Dott. Giuseppe Buttiglione
