ATHEROGENIC BIOMARKERS IN VASCULAR DEMENTIA:

THE ROLE OF INFLAMMATION IN NEURODEGENERATION

J.C. Carril and R. Cacabelos
· Institute for CNS Disorders and Genomic Medicine, EuroEspes Biomedical Research Center, Corunna, Spain ·
· Chair of Genomic Medicine, Camilo Jose Cela University, Madrid, Spain ·

BACKGROUND RESULTS
The molecular basis for the genetic risk of cerebrovascular disease is likely to be The most informative genotypes in vascular dementia were IL6*-573GG (29.70%
polygenic and influenced by environmental factors. The influence of cytokine vs. 68.29%, p<0.001) and IL6R*1510AA (25.74% vs. 45.12%, p<0.001). Relevant
gene polymorphisms in modulating the inflammatory pattern involved in forming genotypes for VaE were IL6*-573GG (29.70% vs. 51.67%, p<0.001) and
clots that could trigger arterial ischemic processes has been postulated. IL6R*1510AA (25.74% vs. 45.00%, p<0.001) (Figures 2 and 3).
The ability to identify high-risk patients through genetic testing could make
screening for treatable intermediate phenotypes more cost-effective. Clinical characteristics rs1143634
1,00 0,70
0,60
0,80
0,50
OBJECTIVE 0,60
CON
0,40
CON
VD VD
The aim of this study was to evaluate the role of specific SNPs in the interleukin 0,40 0,30
VaE VaE
0,20
genes IL1B (C3954T), IL6 (G-174C and G-573C), IL6R (A1510C) and TNF (G-308A) in 0,20 AD
0,10
AD

vascular dementia and other cerebrovascular-related pathologies. 0,00 0,00

Women Obesity HTA HypercholHypertrigl IL1B*3954TT IL1B*3954TC IL1B*3954CC

rs1800795 rs1800796
0,60 1,00
Table 1. Genetic polymorphisms analyzed. (MAF: Minimum Allele Frequency obtained from 0,50 0,80
1000genomes database). 0,40 CON CON
0,60
0,30 VD VD
Symbol Gene name Locus Polymorphism dbSNP Nucleotide change MAF VaE 0,40 VaE
0,20
IL1B interleukin 1, beta 2q13 C3954T rs1143634 c.315C>T T=0.1455 0,10
AD 0,20 AD

IL6 interleukin 6 7p15.3 G-174C rs1800795 c.-237G>C C=0.1850 0,00 0,00

IL6*-174GG IL6*-174GC IL6*-174CC IL6*-573GG IL6*-573GC IL6*-573CC
IL6 interleukin 6 7p15.3 G-573C rs1800796 c.-636G>C C=0.2900
IL6R interleukin 6 receptor 1q21.3 A1510C rs2228145 c.1073A>C C=0.3190
rs2228145 rs1800629
TNF tumor necrosis factor 6p21.3 G-308A rs1800629 c.-488G>A A=0.0960 0,60 1,00
0,50 0,80
0,40 CON CON
0,60
0,30 VD VD
VaE 0,40 VaE
0,20
AD 0,20 AD
0,10
0,00 0,00
DESIGN IL6R*1510AA IL6R*1510AC IL6R*1510CC TNFA*-308GG TNFA*-308GA TNFA*-308AA

A total of 5 genetic variants in 4 different inflammation-related genes (Table 1)

were determined in 652 individuals, 256 patients with vascular dementia (VD), 157 Figure 2. Clinical characteristics and genotype frequencies of Healthy controls (CON), Vascular Dementia
with vascular encephalopathy (VaE), 171 patients (AD) and 110 control subjects patients (VD), Vascular Encephalopathy patients (VaE) and Alzheimer patients (AD).
without ischemic or hemorrhagic stroke history, or other cerebral diseases, aged
over 50 years (Table 2).

OBESITY HYPERTENSION HYPERCHOLESTEROLEMIA

Table 2. Clinical characteristics of patients involved in the study.
Healthy Subjects AD Patients VaD Patients VaE Patients
No. of subjects 68 171 256 157
VD VaE AD VD VaE AD VD VaE AD
Age (years) 56.52 ±11.64 72.88 ± 9.25 74.23 ± 7.98 69.74 ± 9.92
Sex (male/female, %) 38.2 / 61.8 35.1 / 64.9 38.7 /61.3 50.9 / 49.1
Obesity (BMI>25, %) 75.4 70.7 68.0 82.3 IL1B*3954T/C IL1B*3954T/C IL1B*3954T/C
CC, TC CC *C
Hypertension (%) 32.8 34.7 43.3 50.3
Hypercholesterolemia (%) 40.0 30.9 35.2 37.8
Hypertriglyceridemia (%) 15.4 11.3 24.5 30.1
VD VaE AD VD VaE AD VD VaE AD

Genotyping was performed using TaqMan® Assays (Life Technologies) (Figure 1).
IL6*-174G/C IL6*-174G/C IL6*-174G/C
DNA was extracted from whole blood using Qiagen DNA extraction columns. GG, GC GG *G

The standard chi-square method was used to test deviation from the Hardy-
Weinberg equilibrium (HWE) and to compare genotypic and allelic frequencies in
vascular and control groups. Additionally, odds ratios were calculated, and their VD VaE AD VD VaE AD VD VaE AD
appropriate confidence intervals were computed at the 95% level.

IL6*-573G/C IL6*-573G/C IL6*-573G/C

GG, GC GG *G

rs2228145

VD VaE AD VD VaE AD VD VaE AD

0.9
IL6R*1510A/C IL6R*1510A/C IL6R*1510A/C
AA, AC AA *A

0.7 VD VaE AD VD VaE AD VD VaE AD

TNFA*-308G/A TNFA*-308G/A TNFA*-308G/A

AA, GA AA *A

0.5

VD VaE AD VD VaE AD VD VaE AD

0.3
Figure 3. Systemic and genetic factors odd ratios (OR) for Vascular dementia (VD), Vascular
encephalopathy (VE), and Alzheimer disease (AD).

CONCLUSIONS
0.1
The results found in this preliminary study show the relationship between the
plasma levels of cytokines and the major incidence of cerebrovascular accidents.
Increased levels of IL6 and its receptor IL6R increase the recruitment of monocytes
0.1 0.3 0.5 0.7 0.9 1.1 1.3 1.5 1.7
and macrophages to the lesion-prone sites, contributing to the formation of the
A atheroma plaque.
We confirmed a strong association with IL6 and IL6R variants in our patients.
Legend Polymorphisms for this panel of cytokine genes are a valuable tool to assess the
Homozygous A/A Homozygous C/C Homozygous A/C Undeterminated risk of vascular dementia and to help us identify high-risk profiles.

ACKNOWLEDGEMENTS
Figure 1. Allelic discrimination plot from TaqMan® OpenArray® DNA Genotyping platform. This investigation was partially supported by a grant from the Camilo José Cela
University (Genescreen project 2014-2015).