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
