A Gata456 Pipeline of Discovery

三个高度相关的基因，Gata4，Gata5和GATA6(这里称为Gata456)基本上调节每一个 心脏生物学方面，从心前中胚层的产生、特化和分化 心内膜、心外膜和心肌祖细胞，心管形成，生长和形态发生， 间隔和瓣膜形成，心脏保护和肥大，以及再生。这三个基因是如何 调控空间、时间和组织特有的遗传和表观遗传网络，这些网络是所有这些的基础 人们对不同的项目知之甚少。此外，每个基因的突变都是单独发生的 与人类心肌病有关，包括房间隔和室间隔缺陷，法洛四联症， 二尖瓣主动脉瓣综合征和家族性扩张型心肌病。其他转录因子基因，以及 已知一些末端分化标记受Gata456调控，但在理解上存在重大差距 是识别控制中介功能的关键目标基因，如谱系指定、生长、 形态发生和心脏保护。我们提出了一个新的计划，作为一条“发现管道”，以识别 这些下游基因及其在心脏发生和心脏生物学中的功能。总体目标是 确定每个Gata456基因在不同心脏组织发育和成人生活中的功能 包括心内膜、心肌和心外膜。我们试图破译的密码是如何相对定时 每个基因的表达和位置影响细胞的命运和存活，以及器官的形态发生和 功能。互补的模式系统利用特定的优势并解决特定物种的差异： 用于了解心脏发生包括形态发生的斑马鱼，以及用于 了解人类细胞身份和疾病模型。我们编辑了一个斑马鱼和hESC的“工具箱” LINES和一个调查专家团队，以促进对功能得失的全面分析 表型，在此类分析和发现新的下游靶标方面有良好的记录。一个 要了解Gata456如何控制心脏生成的各个方面，需要取得突破。我们是 最后通过对各个因素在不同发展阶段的系统操作，来定义这一编码 和组织背景，导致发现特定的关键下游靶基因，这些基因执行这些多样化 功能。该项目不会直接开发心脏病的治疗方法，但它可能会增强 细胞疗法的发展。主要是，它将突破目前对监管的描述 两个领域的网络：1)定义单个Gata456等位基因在特定情况下对损失或获得的影响 在发育阶段和特定组织中精确定义发育动物(斑马鱼)的功能 和人类细胞(来源于人类多能细胞)。2)确定关键下游Gata456目标 负责阶段和组织特定功能的基因，识别这些是“铅中毒”的治疗方法 治疗心脏病的目标。 好了！

Three highly related genes, Gata4, Gata5, and Gata6 (referred to here as Gata456) regulate essentially every aspect of cardiac biology, from generation of precardiac mesoderm, specification and differentiation of endocardial, epicardial, and myocardial progenitors, heart tube formation, growth and morphogenesis, septation and valve formation, cardioprotection and hypertrophy, and regeneration. How the three genes regulate the spatial, temporal, and tissue-specific genetic and epigenetic networks that underlie all of these disparate programs is poorly understood. Furthermore, mutations in each of the genes have individually been associated with human cardiomyopathies, including atrial and ventricular septal defects, tetralogy of Fallot, bicuspid aortic valve syndrome, and familial dilated cardiomyopathy. Other transcription factor genes, and some terminal differentiation markers are known to be regulated by Gata456, but a major gap in understanding is the identify of the key target genes that control intermediary functions such as lineage specification, growth, morphogenesis, and cardio-protection. We propose a new program as a “Pipeline of Discovery” to identify these downstream genes and probe their function in cardiogenesis and cardiac biology. The overall goal is to define the function of each Gata456 gene throughout development and adult life in various cardiac tissues including endocardium, myocardium, and epicardium. We seek to break the code for how the relative timing and location of expression for each gene impacts cell fate and survival, and organ morphogenesis and function. Complementary model systems exploit specific advantages and resolve species-specific distinctions: the zebrafish for understanding cardiogenesis including morphogenesis, and human pluripotent stem cells for understanding human cell identity and disease modeling. We have compiled a “toolbox” of zebrafish and hESC lines and an expert team of investigators to facilitate a comprehensive analysis of gain-and loss-of-function phenotypes, with a strong track record for such analyses and discovery of novel downstream targets. A breakthrough is needed to understand how Gata456 controls all the various aspects of cardiogenesis. We are finally in a position to define this code, by a systematic manipulation of each factor in different developmental and tissue contexts, leading to discovery of specific key downstream target genes that carry out these diverse functions. This project will not directly develop therapeutics for cardiac disease, but it will likely enhance development of cellular therapies. Chiefly, it will break ground beyond current descriptions of regulatory networks in two areas: 1) Defining the impact for loss or gain of individual Gata456 alleles at specific developmental stages and in specific tissues to precisely define functions in developing animals (zebrafish) and human cells (derived from human pluripotent cells). 2) Identifying the key downstream Gata456 target genes that are responsible for stage and tissue-specific functions, recognizing these as “lead hit” therapeutic targets for treating cardiac disease. !