GENOMIC WIDE ASSOCIATION STUDY FOR

HYPERTENSION
ZENOVIA ANDREI1
1
Lucian Blaga University, Faculty of Medicine
………………………………………………………………………………….

* Corresponding author: zenovia.andrei@gmail.com

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1. Introduction
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2. Materials and Methods

2.1. Study population
Subjects included in this study were male and female patients admitted between 2008 and 2012 to two
clinics in Bucharest Urology Clinic ‘Th. Burghele’ and General Surgery Clinic „Sf. Maria” for various
medical conditions. All subjects gave written informed consent prior to enrolment and accepted the use
of personal and clinical data and biological samples for genetic research. The Bioethical Committee of
the Romanian College of Physicians approved the study and the study protocols were approved by the
National Ethical Board of the Romanian Medical Doctors Association in Romania. Trained
interviewers performed face-to-face interviews, using standardized questionnaires, to collect personal
data (ethnicity, marital status, education, height and weight), lifestyle data (occupation, smoking,
coffee and tea consumption) and medical history (personal and familial). All subjects were of self-
reported European descent. No significant difference was observed between the average age of the
cases (66.9) and controls (64.3)
2.2. Genotyping and analysis of Single-Nucleotide Polymorphism (SNP) data
DNA was extracted from whole blood at deCODE Genetics (Reykjavik, Iceland) and genotyped using
Infinium OmniExpress-24 bead chips (Illumina). A total of 716,503 SNPs were genotyped for each
individual included in the study. The genotype data were filtered using Plink! v1.07 15. Approximately
10% of the SNPs genotyped were removed using a Hardy–Weinberg equilibrium significance
threshold of 5 × 10-6 and by excluding markers with a minor allele frequency lower than 1%. Prior to
the imputation, each chromosome was phased in a single run using SHAPEIT 16. Markers from Phase
3 October 2014 of the 1000 Genomes 17 were imputed into the 2024 chip-typed individuals using the
IMPUTE2 software 18 with a posterior probability of 0.9 as a threshold to call genotypes. The set of
genotypes were tested for population heterogeneity using principal component analysis in the
ADMIXTURE software 19, and the results were consistent with a homogeneous population.

3. Results and Discussions (Head 3 – 14 Bold)

After an extensive search of the known literature, we have decided on the following as susceptible
genes that should be further studied in order to prove a link with hypertension.

3.1. GWAS Analysis

We report here a genome-wide association study (GWAS) for a complex trait, namely hypertension.
We performed an initial GWAS in cohort of the ROMCAN project combining data collected from
5385 individuals 78% males of Romanian descent out of which 38% where cases and 60% controls.
We identified 24 loci out of these 5 strongly associate with hypertension. Using bioinformatics, we
show that identified genes are enriched for expression in heart muscle myocytes and in pathways
relevant for hypertension, transmembrane transporter activity, arterial wall structure and regulation of
Ca2+ metabolism.
We explored the different complex physiopathology mechanisms that are known to cause hypertension
and then associated the identified loci with individual pathologic pathways that may cause the disease,
and then cross referenced our findings with published results of experimental work revolving around
this gene.
3.2. Genes with a strong association with hypertension
The gene ITPR2 is located on chromosome 12 has as its main function the coding for the receptor for
inositol 1,4,5-trisphosphate, a second messenger that mediates the release of intracellular calcium.
ITPR2 has been shown to affect nutrient metabolism and central obesity. Dietary patterns, genetic
variants, and host characteristics. (Melkonian) Experimental reserch on transgenic mice with the high-
expressing IP3 receptor mild cardiac hypertophy by 3 months of age , and consistent with this
phenotype the reexpression of the hypertrophy -related genes atrial natriuretic factor and skeletal α-
actin were observed in the hearts of high-expressing IP3R mice, and no cardiac hypertrophy in the low-
expressing IP3 transgenic mice. (Nakayama)

3.3. Figures and Tables

For this table we have used the SNPs with a p-value of ~ 0.05 resulted from the data analysis with the
plink software and RStudio. After the identification of the SNPs, we conducted a search using the
National Center for Biotechnology Information and dbSNP database. After we discovered a link
between our SNPs and a GENE, we then conducted a search for the respective GENE in the
GeneCards®: The Human Gene Database and after we identified a link between the GENE and our
test hypothesis, we then conducted a search on the GWAS Catalog for further literature directed
review. We then searched for the genetic databases for signals of GENE expression in various human
tissues in order to discover any possible link to hypertension.

Table 1. Single-Nucleotide Polimorphism association with GWAS for Hypertension

Group Chromosome SNPs GENE Link to HTA
A 1 rs6657631 - Unknown
 Group Chromosome SNPs GENE Link to HTA
rs2454228 - Unknown
rs6720101 PANTR1 BMI
2 rs10932355 - Unknown
rs11695055 DNER BMI
rs4684862 CNTN6 DBP
rs9845983 THRB Erythrocyte count
3
rs12489864 EPHA6 SBP
rs7621659 CPNE4 BMI
4 rs10013360 CCSER1 Unknown
B
5 rs891980 - Unknown
rs9493826 - Unknown
6 PDE7B Erythrocyte count
rs7747433
LOC644135 Intraocular pressure
Glomerular
7 rs10480299 PRKAG2
filtration rate
rs10755966 ZNF704 Unknown
8
rs1378900 LOC105375752 Waist/Hip ratio
9 rs7855735 LOC105376234 Brain Aneurysm
C
10 rs11253143 - Unknown
rs4539314 LRRC4C Unknown
Renal Carcinoma
rs640908 PLAAT3
11 Antigen
rs10502138 - Unknown
rs12799000 - Unknown
rs12368682 - Unknown
12
rs2230377 ITPR2 BMI; Waist / Hip
rs10507431 DCLK1 Unknown
13 Chronic Kidney
rs12585074 NALCN-AS1
Disease
D BMI
14 rs12435483 TRAF3
15 rs16952930 - Unknown
16 rs7505073 - Unknown
17 rs16952930 - Unknown
E
18 rs7505073 - Unknown
rs2305758 HAUS8 SBP; CVD
19
rs8104920 ZNF836 Unknown
F
rs1111036 LOC107985399 Unknown
20
rs6025161 - Unknown
21 rs914186 BACE2 BMI
G 0,3
22 rs3788310 TXNRD2 AKI
y = 0.0001x
0,25Diastolic blood
BMI: Body Mass Index; DBP: pressure;+SBP:
0.1678
Systolic blood pressure; CVD: Cardio Vascular Disease AKI: Acute
R2 = 0.9995Kidney injury
0,2
660 nm

0,15

0,1

0,05

0
0 0,001 0,002 0,003 0,004 0,005 0,006
concentration, g/L
 Figure 1. Example of a figure caption (figure caption).
3.3. ……………..

4. Conclusions (Head 4 – 14 Bold)

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References (Head 5 – 12 Bold)

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6. Title of website. Available online: URL (accessed on Day Month Year)
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Manuscript received: 11.10.2019
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