DOI: 10.5061/dryad.bg79cnpfx
title: Polymorphisms in genes related to inflammation and endothelial function in the high-risk stroke population

description: Aim: To investigate incidence of ischemic stroke and other vascular events
 during a 4.7-year follow-up in the high-risk stroke population,
 and to identify the associations of the 19 single nucleotide polymorphisms
 (SNPs) in genes related to inflammation and endothelial
 function and interaction among these SNPs with outcomes.  
 Methods: According to the China National Stroke Screening Survey
 programme, we performed this multi-center community-based sectional survey
 and prospective cohort study in the Sichuan of southwestern China from May
 2015 to January 2020. Eight communities were randomly selected in Sichuan,
 and the residents in each community volunteered to participate in
 a face-to-face survey. The 19 SNPs in genes related to endothelial
 function and inflammation were measured in the high-risk stroke
 population. All subjects in high-risk stroke populations were followed-up
 for 4.7 years after the face-to-face survey. The primary outcome was a new
 ischemic stroke; the secondary outcome was a composite of new vascular
 events.  Results: A total of 2893 high-risk stroke population,
 2698 (93.3%) completed a 4.7-year follow-up. Outcomes occurred in 192
 (7.1%) subjects (118 [4.4%] new ischemic stroke, 24 [0.9%] hemorrhagic
 stroke, 53 [2.0%] myocardial infarction, and 33[1.2%] death) in the 2698
 high-risk stroke population. There were significant differences in
 genotype distribution of TNF rs3093662, IL6R rs4845625 and TLR4 rs752998
 between subjects with and without outcomes by univariate analyses.
 Generalized multifactor dimensionality reduction (GMDR) analysis showed
 that there was a significant gene-gene interaction among the 19 SNPs, the
 best model for outcomes was interaction among IL6R rs4845625, TLR4
 rs1927911 and HABP2 rs932650 (P = 0.004). The high-risk interactive
 genotypes among the 3 SNPs were independently associated with a higher
 risk for new ischemic stroke  (OR = 2.187, 95% CI: 1.256–5.374,
 P＜0.001) and total vascular events (OR = 2.382, 95% CI: 1.423–5.894,
 P＜0.001) after adjustment with covariates. Conclusion: The incidence of
 ischemic stroke and other vascular events was found to be very high in the
 high-risk stroke population. There were associations of specific SNPs in
 genes related to inflammation and endothelial function
 with outcomes. The high-risk interactive genotypes among IL6R rs4845625,
 TLR4 rs1927911 and HABP2 rs932650 were independently associated with a
 higher risk for new ischemic stroke and other vascular events. These
 findings are expected to provide new strategies for prevention of ischemic
 stroke and other vascular events.
description: Study design and
 participants  According to the CNSSS
 programme, we performed this multi-center community-based cross-sectional
 survey and prospective cohort study in Sichuan of southwestern China from
 May 2015 to January 2020. The survey protocol was reviewed and
 approved by the Ethics Committee of participating hospitals and informed
 consent (in the Chinese language) was obtained from all participants
 before enrollment. The community-based cross-sectional survey
 was conducted in 8 communities of Sichuan between May 2015 and September
 2015. The detailed procedures for recruitment of
 participants and evaluation of risk factors were described in our previous
 articles. In brief, the 8 communities were selected
 using cluster randomization method in Sichuan. The residents aged ≥
 40 years in each community were screened using a structured face-to-face
 questionnaire by interviewers. The questionnaire included demographic
 characteristics, behavioural factors, history of chronic diseases,
 personal and family medical history of stroke, and physical
 examination. Evaluation of risk
 factors and definition of high-risk population for stroke
 Information on
 demographic characteristics was collected, and the eight conventional risk
 factors for stroke and history of stroke were evaluated.
 The eight risk factors included
 overweight/obesity, smoking, physical inactivity, family history of
 stroke, hypertension, diabetes mellitus, dyslipidemia, and atrial
 fibrillation.  The detailed diagnostic criteria for the
 eight risk factors and stroke were described in our previous article.
 
 Subjects with at least three of the aforementioned eight risk factors or a
 history of stroke were defined as the high-risk stroke population.
 
 The exclusion criteria were as follows: (1) residents declined to
 participate in the study; (2) severe cardiovascular, liver or renal
 disease, blood system disease, acute and chronic inflammation, malignant
 tumors, and immune system diseases;  (3) history of
 carotid artery stenting or
 endarterectomy. Genotyping
 The 19 SNPs in 10
 genes related to inflammation and endothelial function were selected from
 the NCBI database (http://www.ncbi.nlm.nih.gov/SNP), according to the
 following criteria: (1) the SNPs have been evaluated in previous studies
 ;  (2) the SNPs have a minor allele frequency
 &gt; 0.05; (3) the SNPs may lead
 to amino acid changes. For the high-risk stroke population,
 peripheral blood (3 mL) was drawn from an arm vein and was stored in
 cryogenic refrigerator at -80 °C until genotype analysis was performed.
  Genomic DNA was extracted by a modified
 phenol/chloroform method and purified using a UNIQ-10 kit (Sangon Biotech
 Co., Ltd., Shanghai, China). Genotypes of the 19 SNPs were evaluated
 using a matrix-assisted laser desorption/ionization time of flight mass
 spectrometry (MALDI-TOF MS) method by investigators, as were previously
 described.  The investigators were blinded to the data
 of participants.
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