Defining the role of Pou6f1 in cardiac morphogenesis

确定Pou6f1在心脏形态发生中的作用 项目总结 先天性心脏病(CHD)是导致出生缺陷的主要原因，并长期存在 尽管当代管理有了明显的改善，但仍有预后。这一失败源于严重的LIM- 智能诊断能力，再加上完全无法修改潜在的疾病过程。Alt- 尽管负责心脏发育的转录途径已经有了很好的特征，但新的 迫切需要已建立的电路的调节器来识别新的CHD候选基因并阐明 潜在的治疗靶点。这项研究计划的长期目标是了解转录连接。 以房室管(AVC)分化为模型系统的心脏谱系规范。这个 该提案的目的是描述最近确定的AVC规范的上游调节器的特征。我们的 中心假说是Pou6f1调节心脏结构和功能。为了检验这一假设，我们建议 具体目标如下：1)分析Pou6f1过表达和Pou6f1缺失导致的心脏表型。 终止Pou6f1缺失对心功能的影响。我们的初步数据显示心脏特异性 显性负性Pou6f1的过度表达扰乱了AVC的形态发生，并导致胚胎死亡。 在目标1中，我们将检查Pou6f1功能获得在心脏发育过程中的后果，通过使用 我们在实验室中培育了两个组织特异性Pou6f1过表达的小鼠系。额外的Pre- 初步研究结果表明，全局Pou6f1缺失会导致男性特有的胚胎死亡和心肌病 在幸存的雌性中。在目标2中，我们将评估全局和心脏特异性的功能后果 Pou6f1缺失使用我们先前获得的花序等位基因。我们的方法是创新的，因为它 将挑战和扩展现有的AVC形态发生范式，这是正确形成AVC的关键一步 房间隔、室间隔和房室瓣。因此，这个项目意义重大，因为它将意味着- 在AVC规范和心脏形态发生中创建一个全新的转录途径。带到一起- 呃，拟议研究的预期好处是确立Pou6f1在心脏形态发育过程中的作用。 Genesis，并为竞争性R01提交获得关键的初步数据。我们预计随后会出现 后续研究将对AVC的形态发生机制有更深入的了解。反过来, 这些见解有望扩大候选基因座的名单，并为小说的合理设计提供平台 动静脉曲张的治疗。

Defining the role of Pou6f1 in cardiac morphogenesis PROJECT SUMMARY Congenital heart disease (CHD) is the leading cause of birth defects and continues to carry a poor long-term prognosis despite marked improvements in contemporary management. This failure stems from severely lim- ited diagnostic capabilities coupled with a complete inability to modify the underlying disease process. Alt- hough the transcriptional pathways responsible for heart development have been well-characterized, new regulators of the established circuitry are urgently needed to identify novel CHD candidate loci and illuminate potential therapeutic targets. The long-term goal of this research program is to understand transcriptional con- trol of cardiac lineage specification using atrioventricular canal (AVC) differentiation as a model system. The objective of this proposal is to characterize a recently identified upstream regulator of AVC specification. Our central hypothesis is that Pou6f1 regulates cardiac structure and function. To test this hypothesis, we propose the following specific aims: 1) Analyze cardiac phenotypes resulting from Pou6f1 over-expression and 2) De- termine the impact of Pou6f1 deletion on cardiac function. Our preliminary data indicate that cardiac-specific over-expression of dominant-negative Pou6f1 perturbs AVC morphogenesis and results in embryonic lethality. In Aim 1, we will examine the consequences of Pou6f1 gain-of-function during heart development by utilizing two mouse lines for tissue-specific Pou6f1 over-expression that we have generated in our lab. Additional pre- liminary results show that global Pou6f1 deletion causes male-specific embryonic lethality and cardiomyopathy in surviving females. In Aim 2, we will evaluate the functional consequences of global and cardiac-specific Pou6f1 deletion using a floxed allele that we have previously obtained. Our approach is innovative because it will challenge and expand the existing paradigm for AVC morphogenesis, a critical step for proper formation of the atrial septum, ventricular septum, and AV valves. This project is significant, therefore, because it will impli- cate an entirely new transcriptional pathway in AVC specification and cardiac morphogenesis. Taken togeth- er, the expected benefits from the proposed studies are to establish a role for Pou6f1 during cardiac morpho- genesis and to obtain key preliminary data for a competitive R01 submission. We anticipate that subsequent follow-up studies will provide a more informed mechanistic understanding of AVC morphogenesis. In turn, such insights promise to expand the list of candidate loci and provide a platform for the rational design of novel treatments for AVSDs.