Supplementary Material for: Analysis of rare variants in 470,000 exome-sequenced UK Biobank participants implicates novel genes affecting risk of hypertension

Introduction A previous study of 200,000 exome-sequenced UK Biobank participants to test for association of rare coding variants with hypertension implicated two genes at exome-wide significance, DNMT3A and FES. A total of 42 genes had an uncorrected p value < 0.001. These results were followed up in a larger sample of 470,000 exome-sequenced participants. Methods Weighted burden analysis of rare coding variants in a new sample of 97,050 cases and 172,263 controls was carried out for these 42 genes. Those showing evidence for association were then analysed in the combined sample of 167,127 cases and 302,691 controls. Results The association of DNMT3A and FES with hypertension was replicated in the new sample and they and the previously implicated gene NPR1, which codes for a membrane bound guanylate cyclase, were all exome-wide significant in the combined sample. Also exome-wide significant as risk genes for hypertension were GUCY1A1, ASXL1 and SMAD6, while GUCY1B1 had a nominal p value of < 0.0001. GUCY1A1 and GUCY1B1 code for subunits of a soluble guanylate cyclase. For two genes, DBH, which codes for dopamine beta hydroxylase, and INPPL1, rare coding variants predicted to impair gene function were protective against hypertension, again with exome-wide significance. Conclusion The findings offer new insights into biological risk factors for hypertension which could be the subject of further investigation. In particular, genetic variants predicted to impair the function of either membrane-bound guanylate cyclase, activated by natriuretic peptides, or soluble guanylate cyclase, activated by nitric oxide, increase risk of hypertension. Conversely, variants impairing the function of dopamine beta hydroxylase, responsible for the synthesis of norepinephrine, reduce hypertension risk. This research has been conducted using the UK Biobank Resource.

引言
此前一项针对20万名接受外显子组测序的英国生物库（UK Biobank）参与者开展的研究，旨在探究罕见编码变异与高血压的关联，在外显子组水平显著性标准下鉴定出两个相关基因：DNMT3A与FES。共计42个基因的未校正p值小于0.001。上述研究结果在更大规模的47万名接受外显子组测序的参与者队列中得到了后续验证。

方法
针对上述42个基因，在新纳入的97050例病例与172263例对照样本中开展罕见编码变异的加权负担分析。对显示出关联证据的基因，在合并后的167127例病例与302691例对照样本中进行进一步分析。

结果
DNMT3A与FES和高血压的关联在新队列中得到了重复验证；在合并样本中，这两个基因以及此前被提示关联的基因NPR1（编码膜结合鸟苷酸环化酶（membrane bound guanylate cyclase））均达到了外显子组水平显著性。另外，GUCY1A1、ASXL1与SMAD6作为高血压风险基因也达到了外显子组水平显著性；而GUCY1B1的名义p值小于0.0001。GUCY1A1与GUCY1B1分别编码可溶性鸟苷酸环化酶（soluble guanylate cyclase）的亚基。针对另外两个基因DBH（编码多巴胺β羟化酶（dopamine beta hydroxylase））与INPPL1，研究发现预测会损伤基因功能的罕见编码变异可对高血压起到保护作用，且该关联同样达到了外显子组水平显著性。

结论
本研究结果为高血压的生物学风险因素提供了新的见解，可作为后续研究的方向。具体而言，预测会损伤由利钠肽激活的膜结合鸟苷酸环化酶，或是由一氧化氮激活的可溶性鸟苷酸环化酶功能的遗传变异，会升高高血压发病风险。反之，损伤负责去甲肾上腺素合成的多巴胺β羟化酶功能的变异，则会降低高血压发病风险。本研究使用英国生物库（UK Biobank）资源开展。