title: Novel Thrombotic Function of a Human SNP in STXBP5 Revealed by CRISPR/Cas9 Gene Editing in Mice.
abstract: OBJECTIVE
  To identify and characterize the effect of a SNP (single-nucleotide polymorphism) in the STXBP5 locus that is associated with altered thrombosis in humans. GWAS (genome-wide association studies) have identified numerous SNPs associated with human thrombotic phenotypes, but determining the functional significance of an individual candidate SNP can be challenging, particularly when in vivo modeling is required. Recent GWAS led to the discovery of STXBP5 as a regulator of platelet secretion in humans. Further clinical studies have identified genetic variants of STXBP5 that are linked to altered plasma von Willebrand factor levels and thrombosis in humans, but the functional significance of these variants in STXBP5 is not understood.
  
  
  RESULTS
  We used CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated 9) techniques to produce a precise mouse model carrying a human coding SNP rs1039084 (encoding human p. N436S) in the STXBP5 locus associated with decreased thrombosis. Mice carrying the orthologous human mutation (encoding p. N437S in mouse STXBP5) have lower plasma von Willebrand factor levels, decreased thrombosis, and decreased platelet secretion compared with wild-type mice. This thrombosis phenotype recapitulates the phenotype of humans carrying the minor allele of rs1039084. Decreased plasma von Willebrand factor and platelet activation may partially explain the decreased thrombotic phenotype in mutant mice.
  
  
  CONCLUSIONS
  Using precise mammalian genome editing, we have identified a human nonsynonymous SNP rs1039084 in the STXBP5 locus as a causal variant for a decreased thrombotic phenotype. CRISPR/Cas9 genetic editing facilitates the rapid and efficient generation of animals to study the function of human genetic variation in vascular diseases.
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Qiuyu Martin
authors.initials: QM
authors.lastname: Zhu
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Kyung Ae
authors.initials: KA
authors.lastname: Ko
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Sara
authors.initials: S
authors.lastname: Ture
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Michael A
authors.initials: MA
authors.lastname: Mastrangelo
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Ming-Huei
authors.initials: MH
authors.lastname: Chen
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Johnson
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: O'Donnell
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Craig N
authors.initials: CN
authors.lastname: Morrell
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.).
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Miano
authors.affiliation: From the Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester Medical Center, NY (Q.M.Z., K.A.K., S.T., M.A.M., C.N.M., J.M.M., C.J.L.); Division of Intramural Research, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health, and NHLBI Framingham Heart Study, Framingham, MA (M.-H.C., A.D.J., C.J.O.D.); and Cardiovascular Division, Massachusetts General Hospital, Harvard Medical School, Boston (C.J.O.D.). charles_lowenstein@urmc.rochester.edu.
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Lowenstein
mesh_terms: D000818:Animals; D001777:Blood Coagulation / Q000235:genetics*; D001792:Blood Platelets / Q000378:metabolism; D064130:CRISPR-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004195:Disease Models, Animal; D015536:Down-Regulation; D005089:Exocytosis; D000072669:Gene Editing / Q000379:methods*; D055106:Genome-Wide Association Study; D005838:Genotype; D006801:Humans; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000235:genetics*; D010641:Phenotype; D015539:Platelet Activation; D020641:Polymorphism, Single Nucleotide; D050683:R-SNARE Proteins / Q000235:genetics*; D013927:Thrombosis / Q000097:blood / Q000235:genetics / Q000517:prevention & control*; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D064130:CRISPR-Associated Proteins; D009419:Nerve Tissue Proteins; D050683:R-SNARE Proteins; C484767:STXBP5 protein, human; D014841:von Willebrand Factor
keywords: gene editing; genome-wide association study; phenotype; thrombosis; von Willebrand factor