Reconstruction of cardiovascular regulatory networks from large-scale single-cell analyses of cardiovascular lineages

 描述(申请人提供)：人类早期心血管(CV)发育的基因调控网络知之甚少，这在很大程度上是由于缺乏指定心血管祖细胞类型(CVPC)多样性的分子和遗传信息。人多能干细胞(HPSC)来源的CV细胞为人类心脏发生提供了一个模型，使我们有机会揭示心脏发育过程中产生的各种CV细胞类型，并从功能上发现和验证CV发育基因调控网络。在这项建议中，我们将使用单细胞转录组(RNA-seq)分析来剖析早期CV祖细胞群体的异质性，这些异质性导致了不同的CV细胞类型及其中间产物的光谱。通过识别这些潜在的稀有和新颖的前体细胞类型，以及在单细胞水平上研究它们的谱系选择决定，可以揭示这些前体细胞及其分化的CV细胞类型背后的细胞和分子网络，这些细胞和分子网络控制它们的分化。为了实现我们的目标，Yeo和CHI实验室之间的协同和互补合作将致力于(1)研究体外心脏形成过程中CV细胞亚型的多样性和组织，(2)开发能够提取指定CV谱系亚网络的基因调控程序的新算法，以及(3)研究已识别的CV细胞亚型的功能意义。如果成功，我们将揭示治疗先天性心脏病的途径和细胞类型，这将推进我们治疗先天性心脏病的基本和翻译框架。

 DESCRIPTION (provided by applicant): The gene regulatory networks underlying early human cardiovascular (CV) development is poorly understood, in large part due to the dearth of molecular and genetic information specifying the diversity of cardiovascular progenitor cell-types (CVPCs). Human pluripotent stem cell (hPSC)-derived CV cells provide a model for human cardiogenesis and afford us the opportunity to reveal the various CV cell types generated during heart development and to also functionally discover and validate CV developmental gene regulatory networks. In this proposal, we will employ single cell transcriptome (RNA-seq) analysis to dissect the heterogeneity of early CV progenitor populations that give rise to the spectrum of distinct CV cell types and their intermediates. By identifying these potentially rare and novel progenitor cell types as well as studying their lineage choice decisions at the single cell level, the cellular and molecular networks underlying these progenitor cells and their differentiated CV cell types that control their differentiation can be revealed. To achieve our goal, a synergistic and complementary collaboration between the Yeo and Chi labs will aim to (1) investigate the diversity and organization of CV cellular subtypes during cardiogenesis in vitro, (2) develop novel algorithms that enable the extraction of gene regulatory programs that specify CV lineage sub-networks and (3) investigate the functional significance of identified CV cell subtypes. If successful, we will reveal pathways and cell-types that will advance our basic and translational framework for treating congenital heart disease.