Next-generation DNA sequencing to identify novel genetic risk factors for cerebral vein thrombosis

Thromb Res. 2018 Sep:169:76-81. doi: 10.1016/j.thromres.2018.06.011. Epub 2018 Jun 15.

Abstract

Background: Cerebral vein thrombosis (CVT) is a rare, life-threatening disease affecting one adult per 100,000 per year. Genetic risk factors are deficiencies of the natural anticoagulant proteins antithrombin, protein C, protein S or single nucleotide polymorphisms such as factor V Leiden and prothrombin 20210A. In 20% of patients, the cause of CVT remains unknown.

Aim: To identify novel genetic risk factors for CVT using targeted next-generation DNA sequencing (NGS).

Methods: We investigated 171 CVT patients and 298 healthy controls. Patients were selected using the following criteria: objective diagnosis of CVT, no active cancer. We performed targeted NGS analysis of the protein-coding regions of 734 candidate genes related to hemostasis and inflammation, 150 ancestry informative markers and 28 thrombosis-associated variants.

Results: We identified 3723 common and low frequency variants with minor allele frequency (MAF) >1% in 590 genes. Single variant association testing using logistic regression analysis identified rs8176719 insertion/deletion (indel) variant in the ABO gene associated with CVT (age and sex adjusted OR 2.03; 95% CI 1.52-2.73; P = 2.07 × 10-6; Bonferroni P = 0.008). In addition, we identified 8839 rare variants (MAF ≤ 1%) in 723 genes. Gene-based association analysis of these rare variants using a burden test revealed only a tentative association of non-coding variants located in the F8 locus with CVT.

Conclusion: Targeted NGS identified a common indel variant rs8176719 in the ABO gene. Gene-based tests of association failed to reveal genomic loci with a cumulative burden of rare variants associated with CVT.

Keywords: ABO blood group; Cerebral vein thrombosis; Next-generation DNA sequencing; rs8176719.

MeSH terms

  • ABO Blood-Group System / genetics
  • Adult
  • Case-Control Studies
  • Cerebral Veins / metabolism
  • Cerebral Veins / pathology*
  • Female
  • Gene Frequency
  • Genetic Predisposition to Disease
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • INDEL Mutation
  • Intracranial Thrombosis / genetics*
  • Intracranial Thrombosis / pathology
  • Linkage Disequilibrium
  • Male
  • Middle Aged
  • Polymorphism, Single Nucleotide
  • Sequence Analysis, DNA / methods

Substances

  • ABO Blood-Group System