Identifying Sox family transcription factor partners and targets essential for neural crest formation

识别 Sox 家族转录因子伙伴和神经嵴形成所必需的靶标

• 批准号: 10229492
• 负责人: Elizabeth (Betsy) Schock
• 金额: $ 7.19万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2022-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10229492
• 关键词: Affect; Body Regions; Cells; ChIP-seq; Chimeric Proteins; Chondrocytes; Coupled; Craniofacial Abnormalities; Data; Data Set; Defect; Development; Disease; Ectoderm; Embryo; Embryonic Development; Etiology; Evolution; Family; Gene Expression; Genes; Genetic Transcription; Germ Layers; Grant; Individual; Investigation; Lead; Light; Maintenance; Mass Spectrum Analysis; Mediating; Mesoderm; Molecular; Mutation; Neural Crest; Neural Crest Cell; Neuroglia; Patients; Play; Pluripotent Stem Cells; Population; Property; Protein Microchips; Regulation; Regulator Genes; Role; Specificity; Syndrome; Techniques; Tertiary Protein Structure; Tissues; Validation; Vertebrates; Work; blastocyst; blastomere structure; cell type; craniofacial complex; embryo cell; embryonic stem cell; experimental study; in vivo; insight; loss of function; malformation; neurodevelopment; novel; overexpression; pluripotency; pluripotency factor; stem; stem cell population; stem cells; stem-like cell; transcription factor; Affect; Body Regions; Cells; ChIP-seq; Chimeric Proteins; Chondrocytes; Coupled; Craniofacial Abnormalities; Data; Data Set; Defect; Development; Disease; Ectoderm; Embryo; Embryonic Development; Etiology; Evolution; Family; Gene Expression; Genes; Genetic Transcription; Germ Layers; Grant; Individual; Investigation; Lead; Light; Maintenance; Mass Spectrum Analysis; Mediating; Mesoderm; Molecular; Mutation; Neural Crest; Neural Crest Cell; Neuroglia; Patients; Play; Pluripotent Stem Cells; Population; Property; Protein Microchips; Regulation; Regulator Genes; Role; Specificity; Syndrome; Techniques; Tertiary Protein Structure; Tissues; Validation; Vertebrates; Work; blastocyst; blastomere structure; cell type; craniofacial complex; embryo cell; embryonic stem cell; experimental study; in vivo; insight; loss of function; malformation; neurodevelopment; novel; overexpression; pluripotency; pluripotency factor; stem; stem cell population; stem cells; stem-like cell; transcription factor

Project Summary/Abstract Neurocristopathies are a class of syndromes that are predominately characterized by malformations in the craniofacial complex. These defects are caused by aberrant development of the neural crest (NC), a stem cell population unique to vertebrates. As a means of better understanding the molecular basis for neurocristopathies, a thorough investigation of the various aspects of NC cell development is necessary. One unique property of the NC is their broad developmental potential which grants them the ability to give rise to cell types typically attributed to multiple germ layers (mesoderm and ectoderm). Previous work from my lab has lead to the genesis of the hypothesis that the expanded embryonic potential of the NC can be attributed to the retention of stem cell-like pluripotency in these cells. Still a major unanswered question is how NC cells retain their stem cell-like potential even as neighboring cells undergo lineage restriction. Transcription factors often direct cell fate decisions or maintain cell states. In this proposal, I investigate how Sox transcription factors may regulate NC formation and their ability to maintain a stem cell-like state. SoxB1 factors, a subfamily of Sox transcription factors, are expressed in early pluripotent cells of the embryo (blastula) and help to positively regulate pluripotency in that tissue. In contrast, SoxE factors are absent from the blastula, but are robustly expressed in the NC. Effectively, there is switch in the subfamily of Sox factors that is utilized in two temporally distinct stem cell populations in the embryo. Recent work from my lab has shown that this transition from SoxB1 to SoxE factors is essential for NC formation; however, we have yet to define the mechanisms by which SoxE factors help to promote the formation of NC stem cells. In this proposal, I plan to utilize techniques such as IP-mass spectrometry and ChIP-seq to identify potential transcriptional partners and targets of Sox factors that are required for NC formation and controlling pluripotency in the blastula. Using these datasets and further experimental validation of Sox partner and target candidates, I will identify key similarities and differences between the SoxE and SoxB1 partners and targets in these stem cell populations. This will enhance our understanding of the molecular underpinnings of NC formation and maintenance of stem cell potential. Furthermore, these data will help us to understand why the formation of the NC was accompanied by a switch in the utilization of Sox factor subfamilies. !

项目摘要/摘要 神经科学病是一类综合症，主要以畸形为特征 颅面复合物。这些缺陷是由神经克雷斯特（NC）的异常发育引起的，一种干细胞 脊椎动物独有的种群。作为更好地理解分子基础的一种手段 神经科学病，对NC细胞发育的各个方面进行了彻底的研究。一 北卡罗来纳州的独特财产是其广泛的发展潜力，使他们能够产生 细胞类型通常归因于多个细菌层（中胚层和外胚层）。我实验室的先前工作有 导致假说的起源，即NC的扩展胚胎潜力可以归因于 在这些细胞中保留干细胞状的多能性。仍然是一个主要的未解决的问题是NC细胞如何保留 即使相邻细胞受到谱系限制，它们的干细胞样潜力也是如此。经常转录因子 直接细胞命运决定或维持细胞状态。在此提案中，我研究了Sox转录因子如何 调节NC的形成及其维持干细胞状状态的能力。 SOXB1因子，Sox的亚科 转录因子在胚胎（囊肿）的早期多能细胞中表达，并有助于积极 调节该组织中的多能性。相比之下，囊肿不存在袜子因子，但稳健 在NC中表达。有效地，Sox因子的亚家族有切换，在两个时间上使用了 胚胎中不同的干细胞群体。我实验室的最新工作表明，这种过渡 Soxb1至Soxe因素对于NC的形成至关重要。但是，我们尚未定义的机制 SOXE因素有助于促进NC干细胞的形成。在此提案中，我计划利用此类技术 作为IP-MAS光谱法和ChIP-Seq，以识别潜在的转录伙伴和SOX因子的目标 NC形成和控制胚胎中的多能性是必需的。使用这些数据集并进一步 Sox合作伙伴和目标候选人的实验验证，我将确定关键的相似性和差异 在这些干细胞种群中，Soxe和Soxb1伴侣和目标之间。这将增强我们的 了解NC形成和干细胞电位的维持的分子基础。 此外，这些数据将帮助我们了解为什么NC的形成伴随开关 在使用Sox因子亚家族的利用中。 呢