Translating GWAS-identified loci for cardiac rhythm and rate using an in vivo image- and CRISPR/Cas9-based approach.
Translating GWAS-identified loci for cardiac rhythm and rate using an in vivo image- and CRISPR/Cas9-based approach.
复制标题
使用基于体内图像和 CRISPR/Cas9 的方法翻译 GWAS 识别的心律和心率基因座。
DOI：
10.1038/s41598-020-68567-1
复制
发表时间：
2020
影响因子：
4.6
通讯作者：
denHo
中科院分区：
文献类型：
--
作者：
vonderHeyde,Benedikt;Emmanouilidou,Anastasia;Mazzaferro,Eugenia;Vicenzi,Silvia;Höijer,Ida;Klingström,Tiffany;Jumaa,Sitaf;Dethlefsen,Olga;Snieder,Harold;deGeus,Eco;Ameur,Adam;Ingelsson,Erik;Allalou,Amin;Brooke,HannahL;denHo
全基因组关联研究（GWAS）的荟萃分析确定了8个与心率变异性（HRV）相关的基因座，但这些基因座中的候选基因仍未被表征。我们开发了一种基于图像和CRISPR/Cas9的管道，以系统地表征活斑马鱼胚胎中HRV的候选基因。在来自在平滑肌细胞上转基因表达GFP（Tg[acta 2：GFP]）的鱼的卵中同时靶向六个人类候选基因的九个斑马鱼直向同源物，以可视化跳动的heart.An自动化分析在受精后2天和5天对381个活的完整斑马鱼胚胎中的跳动心房的重复30秒记录突出显示影响HRV（hcn 4andsi：dkey-65j6.2 [KIAA 1755]）的基因;心率（rgs 6和hcn 4）;以及窦房暂停和停止的风险（hcn 4）。暴露于10或25 μM ivabradine（一种HCN的开放通道阻滞剂）24小时，导致受精后5天的HRV呈剂量依赖性升高和心率降低。因此，我们的筛选证实了已建立的心率和节律基因（RGS 6和HCN 4）的作用;表明伊伐布雷定降低了斑马鱼胚胎的心率并增加了HRV，就像在人类中一样;并强调了一种在HRV中发挥作用的新基因（KIAA 1755）。
A meta-analysis of genome-wide association studies (GWAS) identified eight loci that are associated with heart rate variability (HRV), but candidate genes in these loci remain uncharacterized. We developed an image- and CRISPR/Cas9-based pipeline to systematically characterize candidate genes for HRV in live zebrafish embryos. Nine zebrafish orthologues of six human candidate genes were targeted simultaneously in eggs from fish that transgenically express GFP on smooth muscle cells (Tg[acta2:GFP]), to visualize the beating heart.An automated analysis of repeated 30 s recordings of beating atria in 381 live, intact zebrafish embryos at 2 and 5 days post-fertilization highlighted genes that influence HRV (hcn4andsi:dkey-65j6.2 [KIAA1755]); heart rate (rgs6 and hcn4); and the risk of sinoatrial pauses and arrests (hcn4). Exposure to 10 or 25 µM ivabradine—an open channel blocker of HCNs—for 24 h resulted in a dose-dependent higher HRV and lower heart rate at 5 days post-fertilization. Hence, our screen confirmed the role of established genes for heart rate and rhythm (RGS6andHCN4); showed that ivabradine reduces heart rate and increases HRV in zebrafish embryos, as it does in humans; and highlighted a novel gene that plays a role in HRV (KIAA1755).