Cis-regulatory circuitry underlying Hedgehog mediated limb development

Hedgehog 介导的肢体发育的顺式调节电路

• 批准号: 8895206
• 负责人: Steven Alexander Vokes
• 金额: $ 30.89万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8895206
• 关键词: Address; Agreement; Antibodies; Binding; Binding Sites; Biological Models; Cleft Palate; Clinical; Cobalt; Congenital Abnormality; Data; Data Set; Defect; Development; Embryo; Enhancers; Erinaceidae; Foundations; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Genomics; Histones; Holoprosencephaly; Human; In Situ Hybridization; Limb Bud; Limb Development; Limb structure; Maps; Mediating; Pathway interactions; Play; Polydactyly; Process; Processed Genes; Proteins; Publishing; Regulation; Regulator Genes; Regulatory Element; Resources; Role; Scientist; Signal Pathway; Signal Transduction; Solid; System; Testing; Transcriptional Regulation; Transgenic Organisms; base; chromatin immunoprecipitation; histone modification; in vivo; innovation; limb bone; loss of function; research study; response; smoothened signaling pathway; transcription factor

DESCRIPTION (provided by applicant): Defects in the Hedgehog (Hh) pathway underlie a vast spectrum of human birth defects, including holoprosencephaly, cleft palate, and polydactyly. Hh signaling controls transcription by regulating the activity of Gli transcription factors. A thorough understanding of how Gli proteins recognize and activate transcriptional targets is therefore necessary to understand the genes and processes that are misregulated in Hh-associated birth defects. However, virtually nothing is known about how Gli proteins recognize target genes and interact with other transcriptional inputs. The aim of this proposal is to define active Gli enhancers and to determine their interactions in a Hh- responsive cis regulatory network that underlies limb development. In the first approach, we will use an innovative approach to identify Hh-responsive, active enhancers different genetic backgrounds. This data will be used to construct a global gene regulatory network, defining an important set of extragenic regions with the collective potential to mediate the transcriptional effects of the Hh pathway. In a second in vivo-centered approach, we will determine the requirement for additional co-activating factors in the transcriptional regulation of Hh target genes. Finally, we will determine if the Hh and BMP pathways co-regulate target genes, focusing on the regulation the developmentally important gene, Gremlin. This study will transform our understanding of limb development by identifying active Gli enhancers and target genes in the limb, defining enhancer domains and determining co-factors. These will provide an invaluable resource for future studies and will provide a solid foundation for a systems-level understanding of limb development. The identification of these enhancers will also provide an important genomic resource for clinical scientists seeking to understand the mechanisms underlying the spectrum of BMP and Hh-associated birth defects and the hundreds of clinically distinct forms of human polydactyly.

描述（由申请人提供）：Hedgehog (Hh) 通路的缺陷是多种人类出生缺陷的基础，包括前脑无裂畸形、腭裂和多指畸形。 Hh 信号通过调节 Gli 转录因子的活性来控制转录。因此，有必要彻底了解 Gli 蛋白如何识别和激活转录靶标，以了解 Hh 相关出生缺陷中失调的基因和过程。然而，对于 Gli 蛋白如何识别靶基因并与其他转录输入相互作用，几乎一无所知。该提案的目的是定义活性 Gli 增强子并确定它们在肢体发育基础的 Hh 响应顺式调控网络中的相互作用。在第一种方法中，我们将使用创新方法来识别不同遗传背景的 Hh 响应性主动增强子。这些数据将用于构建一个全球基因调控网络，定义一组重要的基因外区域，这些区域具有介导 Hh 途径转录效应的集体潜力。在第二种以体内为中心的方法中，我们将确定 Hh 靶基因转录调控中对额外共激活因子的需求。最后，我们将确定 Hh 和 BMP 途径是否共同调节靶基因，重点关注发育重要基因 Gremlin 的调节。这项研究将通过识别肢体中的活性 Gli 增强子和目标基因、定义增强子域并确定辅助因子来改变我们对肢体发育的理解。这些将为未来的研究提供宝贵的资源，并为对肢体发育的系统级理解提供坚实的基础。这些增强子的鉴定还将为临床科学家提供重要的基因组资源，帮助他们了解 BMP 和 Hh 相关出生缺陷以及数百种临床上不同形式的人类多指畸形的潜在机制。