Abstract
Homocysteine is an independent risk factor for various cardiovascular diseases. There are two ways to remove homocysteine from embryonic cardiac cells: remethylation to form methionine or transsulfuration to form cysteine. Cystathionine β-synthase (CBS) catalyzes the first step of homocysteine transsulfuration as a rate-limiting enzyme. In this study, we identified a functional variant −4673C>G (rs2850144) in the CBS gene promoter region that significantly reduces the susceptibility to congenital heart disease (CHD) in a Han Chinese population consisting of 2 340 CHD patients and 2 270 controls. Individuals carrying the heterozygous CG and homozygous GG genotypes had a 15% (odds ratio (OR) = 0.85, 95% confidence interval (CI) = 0.75-0.96, P = 0.011) and 40% (OR = 0.60, 95% CI = 0.49-0.73, P = 1.78 × 10−7) reduced risk to develop CHD than the wild-type CC genotype carriers in the combined samples, respectively. Additional stratified analyses demonstrated that CBS −4673C>G is significantly related to septation defects and conotruncal defects. In vivo detection of CBS mRNA levels in human cardiac tissues and in vitro luciferase assays consistently showed that the minor G allele significantly increased CBS transcription. A functional analysis revealed that both the attenuated transcription suppressor SP1 binding affinity and the CBS promoter hypomethylation specifically linked with the minor G allele contributed to the remarkably upregulated CBS expression. Consequently, the carriers with genetically increased CBS expression would benefit from the protection due to the low homocysteine levels maintained by CBS in certain cells during the critical heart development stages. These results shed light on unexpected role of CBS and highlight the importance of homocysteine removal in cardiac development.
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Acknowledgements
This work was supported by the grants from the National Science Fund for Distinguished Young Scholars (81025003), the National Basic Research Program of China (973 Program; 2010CB529601), the Program for Innovative Research Team in University (IRT1010), the Doctoral Fund of the Ministry of Education of China (20110071110026), the Commission for Science and Technology of Shanghai Municipality (10JC1401300, 11XD1400900) to HYW, the 973 Program (2012CB910103) to BQ and WYD; and grants from the 973 Program (2012CB944604), the National Natural Science Foundation of China (3100542), the Doctoral Fund of the Ministry of Education of China (20090071120037), and the Natural Science Foundation of Shanghai Municipality (09ZR1404400) to XYY.
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
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Supplementary information, Table S1
The genotype frequency of the 3 identified CBS SNPs in CHD patients and controls (PDF 41 kb)
Supplementary information, Table S2
Demographic characteristics in CHD cases and controls (PDF 68 kb)
Supplementary information, Table S3
DNA sequence of all used primer pairs (PDF 42 kb)
Supplementary information, Figure S1
SPR analysis shows that CBS −4673>A does not change the binding affinity with nuclear extracts. (PDF 54 kb)
Supplementary information, Figure S2
Computational analysis predicted transcription factors for CBS variant −4673C>G and variant −4673>A. (PDF 119 kb)
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Zhao, JY., Yang, XY., Shi, KH. et al. A functional variant in the cystathionine β-synthase gene promoter significantly reduces congenital heart disease susceptibility in a Han Chinese population. Cell Res 23, 242–253 (2013). https://doi.org/10.1038/cr.2012.135
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DOI: https://doi.org/10.1038/cr.2012.135
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