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

项目摘要/摘要 神经管疾病是一类以脑部畸形为主要特征的综合征。 头面部复合体。这些缺陷是由干细胞神经脊(NC)的异常发育引起的 脊椎动物特有的种群。作为更好地理解分子基础的手段 对于神经病变，对NC细胞发育的各个方面进行彻底的调查是必要的。一 NC的独特属性是其广阔的发展潜力，这赋予了他们产生 细胞类型通常由多个胚层(中胚层和外胚层)组成。我实验室以前的工作是 导致假设的起源，即NC的扩展胚胎潜力可以归因于 干细胞样多能性在这些细胞中的保留。仍然是一个主要的悬而未决的问题是NC细胞如何保留 即使邻近细胞受到谱系限制，它们的干细胞样潜能也是如此。转录因子经常 直接决定细胞命运或维持细胞状态。在这个提案中，我研究了SOX转录因子如何 调节NC的形成及其维持干细胞样状态的能力。SoxB1因子，SOX的一个子家族 转录因子在胚胎(囊胚)的早期多能细胞中表达，有助于积极地 调节该组织的多能性。相反，胚泡中不存在SoxE因子，但却非常强健。 在NC中表示。有效地，在两个暂时使用的SOX因子的子家族中存在切换 胚胎中不同的干细胞群体。我的实验室最近的研究表明，从 SoxB1到SoxE因子对于NC的形成是必不可少的；然而，我们还没有定义通过什么机制 SOXE因子有助于促进NC干细胞的形成。在这个提案中，我计划使用如下技术 用IP-MS和CHIP-SEQ鉴定SOX因子的潜在转录伙伴和靶标 这是在囊胚中形成NC和控制多能性所必需的。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，我将找出关键的相似和不同之处 SoxE和SoxB1合作伙伴与这些干细胞群体中的靶点之间的关系。这将增强我们的 了解NC形成和干细胞潜能维持的分子基础。 此外，这些数据将有助于我们理解为什么NC的形成伴随着一个开关 在SOX因子亚家族的利用上。 好了！

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. !