Theis Jeanne L, Vogler Georg, Missinato Maria A, Li Xing, Nielsen Tanja, Zeng Xin-Xin I, Martinez-Fernandez Almudena, Walls Stanley M, Kervadec Anaïs, Kezos James N, Birker Katja, Evans Jared M, O'Byrne Megan M, Fogarty Zachary C, Terzic André, Grossfeld Paul, Ocorr Karen, Nelson Timothy J, Olson Timothy M, Colas Alexandre R, Bodmer Rolf
Cardiovascular Genetics Research Laboratory, Rochester, United States.
Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States.
Elife. 2020 Oct 2;9:e59554. doi: 10.7554/eLife.59554.
Congenital heart diseases (CHDs), including hypoplastic left heart syndrome (HLHS), are genetically complex and poorly understood. Here, a multidisciplinary platform was established to functionally evaluate novel CHD gene candidates, based on whole-genome and iPSC RNA sequencing of a HLHS family-trio. Filtering for rare variants and altered expression in proband iPSCs prioritized 10 candidates. siRNA/RNAi-mediated knockdown in healthy human iPSC-derived cardiomyocytes (hiPSC-CM) and in developing and zebrafish hearts revealed that LDL receptor-related protein is required for cardiomyocyte proliferation and differentiation. Consistent with hypoplastic heart defects, compared to patents the proband's iPSC-CMs exhibited reduced proliferation. Interestingly, rare, predicted-damaging LRP2 variants were enriched in a HLHS cohort; however, understanding their contribution to HLHS requires further investigation. Collectively, we have established a multi-species high-throughput platform to rapidly evaluate candidate genes and their interactions during heart development, which are crucial first steps toward deciphering oligogenic underpinnings of CHDs, including hypoplastic left hearts.
先天性心脏病（CHD），包括左心发育不全综合征（HLHS），具有遗传复杂性，目前人们对其了解甚少。在此，基于一个HLHS家系三联体的全基因组和诱导多能干细胞（iPSC）RNA测序，建立了一个多学科平台，用于对新的CHD基因候选物进行功能评估。通过筛选先证者iPSC中的罕见变异和表达改变，确定了10个候选基因。在健康人iPSC衍生的心肌细胞（hiPSC-CM）以及发育中的斑马鱼心脏中进行小干扰RNA（siRNA）/RNA干扰（RNAi）介导的基因敲低实验，结果显示低密度脂蛋白受体相关蛋白对心肌细胞的增殖和分化是必需的。与心脏发育不全的缺陷一致，与父母相比，先证者的iPSC-CM增殖减少。有趣的是，罕见的、预测具有损害性的LRP2变异在HLHS队列中富集；然而，要了解它们对HLHS的作用还需要进一步研究。总的来说，我们建立了一个多物种高通量平台，以快速评估心脏发育过程中的候选基因及其相互作用，这是破译包括左心发育不全在内的CHD寡基因基础的关键第一步。
