title: Regulation of CARD8 expression by ANRIL and association of CARD8 single nucleotide polymorphism rs2043211 (p.C10X) with ischemic stroke.
abstract: OBJECTIVE
  ANRIL has long been considered as the strongest candidate gene at the 9p21 locus, robustly associated with stroke and coronary artery disease. However, the underlying molecular mechanism remains unknown. The present study works to elucidate such a mechanism.
  
  
  METHODS
  Using expression quantitative loci analysis, we identified potential genes whose expression may be influenced by genetic variation in ANRIL. To verify the identified gene(s), knockdown and overexpression of ANRIL were evaluated in human umbilical vein endothelial cells and HepG2 cells. Ischemic stroke and coronary artery disease risk were then evaluated in the gene(s) demonstrated to be mediated by ANRIL in 3 populations of Chinese Han ancestry: 2 ischemic stroke populations consisting of the Central China cohort (903 cases and 873 controls) and the Northern China cohort (816 cases and 879 controls) and 1 coronary artery disease cohort consisting of 772 patients and 873 controls.
  
  
  RESULTS
  Expression quantitative loci analysis identified CARD8 among others, with knockdown of ANRIL expression decreasing CARD8 expression and overexpression of ANRIL increasing CARD8 expression. The minor T allele of a previously identified CARD8 variant (rs2043211) was found to be significantly associated with a protective effect of ischemic stroke under the recessive model in 2 independent stroke cohorts. No significant association was found between rs2043211 and coronary artery disease.
  
  
  CONCLUSIONS
  CARD8 is a downstream target gene regulated by ANRIL. Single nucleotide polymorphism rs2043211 in CARD8 is significantly associated with ischemic stroke. ANRIL may increase the risk of ischemic stroke through regulation of the CARD8 pathway.
authors.affiliation: From the Key Laboratory of Molecular Biophysics of the Ministry of Education, Cardio-X Institute (Y.B., G.J., Yingchao Zhou, M.Z., Yuanyuan Zhao, S.L., F.W., Q. Lu, Y.H., Q.Y., T.K., H.L., X.R., C.X., X.T., Q.K.W.), and College of Life Science and Technology and Center for Human Genome Research, Institute of Cardiology (S.N., X.C., Y. Liao), Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; Department of Cardiology, Jining Medical College Affiliated Hospital, Jining, China (Q. Li); Department of Cardiology, the First Affiliated Hospital of Harbin Medical University, Harbin, China (Y. Li); Department of Cardiology, First Affiliated Hospital of Dalian Medical University, Dalian, China (Y.X., Y. Liu, J.L., Y.Y.); Department of Cardiology, Suizhou Central Hospital, Suizhou, China (J.Q.); Department of Cardiology, Xiangyang Central Hospital, Xiangyang, China (B.L.); Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, China (G.W.); Department of Cardiology, Wuhan No. 1 Hospital, Wuhan, China (Y.W.); Research Institute of the National Population and Family Planning Commission, Beijing, China (B.W., X.M.); and Department of Molecular Cardiology, Center for Cardiovascular Genetics, Lerner Research Institute, Cleveland Clinic, OH (Q.K.W.).
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authors.forename: Yuhua
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authors.lastname: Liao
authors.forename: Chengqi
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mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D044466:Asian People; D002545:Brain Ischemia / Q000453:epidemiology / Q000235:genetics*; D053468:CARD Signaling Adaptor Proteins / Q000235:genetics*; D002681:China / Q000453:epidemiology; D003324:Coronary Artery Disease / Q000453:epidemiology / Q000235:genetics; D005260:Female; D015870:Gene Expression / Q000502:physiology; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D062085:RNA, Long Noncoding / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D020521:Stroke / Q000453:epidemiology / Q000235:genetics*; D014162:Transfection
chemical_list: D053468:CARD Signaling Adaptor Proteins; C455623:CARD8 protein, human; C561518:CDKN2B antisense RNA, human; D009363:Neoplasm Proteins; D062085:RNA, Long Noncoding
keywords: CARD8 gene; case–control studies; coronary artery disease; polymorphism, single nucleotide; stroke