Unbiased discovery of Novel Regulators of the Cranial Neural Crest

描述(由申请人提供)：这项工作的长期目标是从机制上了解神经脊的发育，神经脊是脊椎动物进化的核心，是一种类似干细胞的前体种群，并了解这种细胞类型的异常发育如何导致先天性畸形和疾病，如癌症。我们试图在机制和生化水平上了解干细胞群体是如何形成的，是什么控制着其多能性的维持，这些细胞的迁移和侵袭行为是如何受到调控的，以及最终，单个神经脊细胞是如何被指示采用特定的衍生命运的。这项提案的总体目标是确定NC-GRN内的新组成部分和相互作用。更具体地说，它试图利用下一代测序来识别神经峰所需的miRNAs和关键神经峰转录因子Snail的下游靶标。我们假设miRNAs在调节神经脊的发育中起着重要的作用，包括它产生头面部骨骼的能力。在神经脊发育过程中表达的miRNAs的特性及其在该组织中的作用在很大程度上仍不清楚。在NC-GRN的核心成分中，Snail转录因子家族已知在其他模式生物的神经脊发育过程中发挥反复作用。因此，这一提议还将通过产生一种以时间控制的方式消融Snail功能的遗传工具，来确定作为Snail功能的结果在神经脊细胞中表达或排除哪些miRNAs。该工具还将用于在全基因组范围内识别Snail在神经脊发育的关键阶段的转录目标。这项工作对于了解包括头面部骨骼在内的神经脊细胞及其衍生物的正常发育，以及了解广泛的与神经脊相关的先天性缺陷具有重要意义。 公共卫生相关性：神经脊(NC)细胞对颅面结构有广泛的贡献，对了解许多先天性疾病具有核心重要性。我们的研究将为调控在NC发育和疾病过程中发挥核心作用的蛋白质提供必要的见解。

DESCRIPTION (provided by applicant): The long-term goal of this work is to develop a mechanistic understanding of the development of the neural crest, a stem cell like progenitor population central to the evolution of the vertebrates, as well as to understand how congenital malformations and diseases such as cancer result from the aberrant development of this cell type. We seek to understand at a mechanistic and biochemical level how the stem cell population is formed, what controls the maintenance of its multipotency, how the migratory and invasive behavior of these cells is regulated, and ultimately, how individual neural crest cells are directed to adopt specific derivative fates. The overall goal of this proposal is to identify novel components and interactions within the NC-GRN. More specifically it seeks to, using next generation sequencing, identify miRNAs required for neural crest and downstream targets of the key neural crest transcription factor Snail. We hypothesize that miRNAs play an essential role in regulating neural crest development, including its ability to give rise to the craniofacial skeleton. The identity of the miRNAs expressed during neural crest development and their roles in this tissue remain largely unknown. Of the core components of the NC-GRN the Snail family of transcription factors is known to play reiterative roles during neural crest development in other model organisms. This proposal will therefore also determine what miRNAS are expressed in or excluded from neural crest cells as a consequence of Snail function by generating a genetic tool for ablating Snail function in a temporally controlled manner. This tool will also be used to identify, on a genome wide scale, Snail transcriptional targets during key stages of neural crest development. This work is of high significance for understanding the normal development of neural crest cells and neural crest derivatives, including the craniofacial skeleton, and for understanding a broad range of neural crest linked congenital defects. PUBLIC HEALTH RELEVANCE: Neural crest (NC) cells make extensive contributions to craniofacial structures and are of central importance to understanding many congenital disorders. Our studies will provide essential insights into the regulation of proteins playing central roles in both NC development and disease processes.