1. Impact of genetic variation in the chemokine system
Possibilities for GENECURE
November
GENECURE
Mike W. Zuurman, PhD

2. Breedtestrategie
Genetic determinants of Atherosclerotic (End-Stage) disease in man
Genetic epidemiology
Patient cohorts
General population
Sequence variants
Functional analyses
In vitro/In vivo
Molecular
Fundamental
Bioinformastatistica
-Utilization of tools
-Development of tools
-Solutions

3. Gene-environment interaction of phenotypic risk factors with
genetic variation in chemokine pathways
CCR2 genotype X Framingham risk score
Impact on CV outcome PREVEND
Impact on CV benefits of antihypertensives (PREVEND)
CCR5 genotype X inflammatory status
Impact on outcome in ESRD (Necosad)

4. CCR2 : G-protein coupled chemokine receptor
CCR2V64I mutation
Valine to isoleucine
Stabilization CCR2A
Isoform:
Impaired downregulation
Associated with
CV disease
Conflicting results

5. Effect of CCR2 on CV events in PREVEND
PREVEND (general population) N=8592
CCR2 genotype: VV/VI/II: 84/15/1 %
CV events : 442 (7 year follow up)
Baseline characteristics similar for genotypes
Framingham risk score (FRS) predicts CV events
FRS: CV events
0-10% : 2.4%
10-20% : 12.3%
20-30% : 19.1%
30% : 26.7%

6. CV hazard ratio by FRS and I-allele
Multivariate hazard analyses:
CCR2 VI+II vs VV : 4.89 (P=0.006)
FRS : 2.20 (P< )
CCR2 * FRS : 1.69 (P=0.005)

7. FRS is a better predictor of CV events in I-carriers
Sensitivity=specificity AUC
83% (VI+II) ( )
73% (VV) ( )

8. CCR2 genotype and CV benefits of antihypertensive treatment
Antihypertensive treatment (AHT)
HR Wald P
AHT ( )
CCR2 mutation ( )
AHT * CCR ( ) <0.0001
Interference with RAAS-blockade ?

9. Conclusions CCR2
Risk of CV morbidity conferred by phenotypic risk factors us
modified by genetic variation in CCR2.
Possible genetic interference with therapeutic efficacy RAAS-blockade
Lit:
AngII induces CCR expression on monocytes
ARB reduces CCR2 expression on monocytes
Possibilities for GENECURE:
Effects of CCR2 mutation in various populations (WP4)
Effects of CCR2 on therapeutic benefit RAAS blockade (WP6)
Pharmaco-economic implications?? (WP9)

10. CCR5 delta32 and inflammation-associated mortality
in ESRD patients (NECOSAD)
CCR5
Involved in atherogenesis and vascular inflammation
CCR5 Δ32:
32bp deletion leads to CCR5 deficiency/dysfunctionality
Associated with improved renal survival in IgA nephropathy
and delayed onset of coronary heart disease in women

11. C-reactive protein in NECOSAD and CCR5
Single or repeated measures of serum CRP are associated with all-cause
and cardiovascular mortality
Hypothesis:
Pro-inflammation
(Micro)inflammation
CRP
CCR5
Mortality

12. CCR5 and inflammation-associated mortality
NECOSAD population
CCR5 wt/wt wt/Δ32 Δ32/Δ32
383 (79.5%) 98 (18.2%) 11 (2.3%)
HW: slight overrepresentation of homozygote Δ32
Dominant model (wt/Δ32 + Δ32/Δ32 are carriers)
Age difference
wt/wt carriers
61.40 ( ) ( )
High CRP defined as CRP>= 10 mg/L

13. Hazard ratios for mortality by CRP and CCR5 genotype
CCR5+CRP Crude overall mortality Adjusted overall mortality
wt/wt & CRP<
wt/wt & CRP> ( ) ( )
carriers & CRP< ( ) ( )
carriers & CRP> ( ) ( )
CCR5 Crude CV mortality Adjusted CV mortality
wt/wt & CRP> ( ) ( )
carriers & CRP< ( ) ( )
carriers & CRP> ( ) ( )

14. Survival for ACM by CRP and CCR5 genotype
Years
wt/wt+<=
carriers+<=
wt/wt+>
carriers+>

15. Conclusion
Association between CRP (marker of inflammation) and CV/overall mortality
depends on CCR5 delta32 genotype.
Pro-inflammation
(Micro)inflammation
CRP
CCR5
Mortality

16. Conclusion/implications
CCR5 delta32 as a protective genetic variant supported by
> Elder age of onset dialysis of carriers
> Slight overrepresentation of homozygote carriers (selection?)
> Better survival despite current inflammation
Treatment with CCR5 antagonists attenuates atherogenesis in
APOE -/- mice !
Possibilities for GENECURE:
Test CCR5 in other populations (WP4)
Test modulation by CCR5 for risk associated with other markers (WP4)
Study (altered) expression of CCR5 in blood vessels (WP2)