TY - JOUR
T1 - A Functional SNP in BNC2 Is Associated with Adolescent Idiopathic Scoliosis
AU - Ogura, Yoji
AU - Kou, Ikuyo
AU - Miura, Shigenori
AU - Takahashi, Atsushi
AU - Xu, Leilei
AU - Takeda, Kazuki
AU - Takahashi, Yohei
AU - Kono, Katsuki
AU - Kawakami, Noriaki
AU - Uno, Koki
AU - Ito, Manabu
AU - Minami, Shohei
AU - Yonezawa, Ikuho
AU - Yanagida, Haruhisa
AU - Taneichi, Hiroshi
AU - Zhu, Zezhang
AU - Tsuji, Taichi
AU - Suzuki, Teppei
AU - Sudo, Hideki
AU - Kotani, Toshiaki
AU - Watanabe, Kota
AU - Hosogane, Naobumi
AU - Okada, Eijiro
AU - Iida, Aritoshi
AU - Nakajima, Masahiro
AU - Sudo, Akihiro
AU - Chiba, Kazuhiro
AU - Hiraki, Yuji
AU - Toyama, Yoshiaki
AU - Qiu, Yong
AU - Shukunami, Chisa
AU - Kamatani, Yoichiro
AU - Kubo, Michiaki
AU - Matsumoto, Morio
AU - Ikegawa, Shiro
N1 - Funding Information:
To examine in vivo effects of BNC2 overexpression, we utilized the tol2-mediated transgenesis system 24,25 in zebrafish. We PCR-amplified the BNC2 coding region from a cDNA clone (IMAGE: 100069207) by using specific primers attached to restriction-enzyme sequences ( Table S4 ). To generate Tol2 transgenesis constructs, p5E-bactin2, pME-MCS BNC2 clone, and p3E-polyA were recombined into pDest-Tol2pA2 by Gateway LR Clonase II enzyme mix (Life Technologies). We co-injected plasmid DNA (10 ng/μl) into one-cell-stage embryos of the RIKEN wild-type strain (Danio rerio, provided by the National BioResource Project of the Ministry of Education, Culture, Sports, Science and Technology, Japan) with capped Tol2 transposase mRNA (50 ng/μl) that was synthesized with a mMESSAGE mMACHINE SP6 Kit (Ambion). We stably expressed BNC2 in zebrafish embryos and analyzed founder transgenic embryos between 24 and 72 hr post-fertilization (hpf). Overexpression of BNC2 in zebrafish embryos resulted in body curvature to various degrees (65%, n = 150) and embryonic lethality (18%, n = 41), whereas EGFP-overexpressing control embryos underwent normal development ( Figures 3 A and 3B ). Most of the abnormal embryos that were delivered with the BNC2 trangene exhibited severe body curvature, and some displayed malformation of the somite, resulting in larval death within one week.
Publisher Copyright:
© 2015 The American Society of Human Genetics.
PY - 2015/8/6
Y1 - 2015/8/6
N2 - Adolescent idiopathic scoliosis (AIS) is the most common spinal deformity. We previously conducted a genome-wide association study (GWAS) and detected two loci associated with AIS. To identify additional loci, we extended our GWAS by increasing the number of cohorts (2,109 affected subjects and 11,140 control subjects in total) and conducting a whole-genome imputation. Through the extended GWAS and replication studies using independent Japanese and Chinese populations, we identified a susceptibility locus on chromosome 9p22.2 (p = 2.46 × 10-13; odds ratio = 1.21). The most significantly associated SNPs were in intron 3 of BNC2, which encodes a zinc finger transcription factor, basonuclin-2. Expression quantitative trait loci data suggested that the associated SNPs have the potential to regulate the BNC2 transcriptional activity and that the susceptibility alleles increase BNC2 expression. We identified a functional SNP, rs10738445 in BNC2, whose susceptibility allele showed both higher binding to a transcription factor, YY1 (yin and yang 1), and higher BNC2 enhancer activity than the non-susceptibility allele. BNC2 overexpression produced body curvature in developing zebrafish in a gene-dosage-dependent manner. Our results suggest that increased BNC2 expression is implicated in the etiology of AIS.
AB - Adolescent idiopathic scoliosis (AIS) is the most common spinal deformity. We previously conducted a genome-wide association study (GWAS) and detected two loci associated with AIS. To identify additional loci, we extended our GWAS by increasing the number of cohorts (2,109 affected subjects and 11,140 control subjects in total) and conducting a whole-genome imputation. Through the extended GWAS and replication studies using independent Japanese and Chinese populations, we identified a susceptibility locus on chromosome 9p22.2 (p = 2.46 × 10-13; odds ratio = 1.21). The most significantly associated SNPs were in intron 3 of BNC2, which encodes a zinc finger transcription factor, basonuclin-2. Expression quantitative trait loci data suggested that the associated SNPs have the potential to regulate the BNC2 transcriptional activity and that the susceptibility alleles increase BNC2 expression. We identified a functional SNP, rs10738445 in BNC2, whose susceptibility allele showed both higher binding to a transcription factor, YY1 (yin and yang 1), and higher BNC2 enhancer activity than the non-susceptibility allele. BNC2 overexpression produced body curvature in developing zebrafish in a gene-dosage-dependent manner. Our results suggest that increased BNC2 expression is implicated in the etiology of AIS.
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U2 - 10.1016/j.ajhg.2015.06.012
DO - 10.1016/j.ajhg.2015.06.012
M3 - Article
C2 - 26211971
AN - SCOPUS:84938969495
SN - 0002-9297
VL - 97
SP - 337
EP - 342
JO - American Journal of Human Genetics
JF - American Journal of Human Genetics
IS - 2
M1 - 1911
ER -