Characterizing the Role of Tspan18 in Neural Crest Development

描述（由申请人提供）：神经嵴是脊椎动物胚胎中一个独特的、迁移的干细胞群体，它产生了许多不同的组织，包括周围神经系统、颅面骨骼、心脏流出道和黑素细胞。神经嵴发育失调引起的大量疾病，包括出生缺陷如口面裂和癌症如神经母细胞瘤，反映了神经嵴衍生物的多样性。尽管神经嵴细胞在发育和疾病中起着关键作用，但控制迁移神经嵴细胞形成的机制尚不清楚。我的初步数据表明，tetraspanin18 （Tspan18）是一种新的、关键的神经嵴迁移特性调节剂。四联蛋白是一种组织和调节细胞信号、运动和粘附的跨膜蛋白。我的研究目的是表征Tspan18在神经嵴发育中的作用。我假设Tspan18是神经嵴细胞膜通过与其他蛋白质相互作用来组织信号的基本支架的一部分，并且Tspan18是神经嵴正常发育所必需的。具体而言，本研究将通过(1)通过反义morpholino寡核苷酸功能缺失分析确定神经嵴对Tspan18的需求，以及(2)通过亲和纯化和质谱分析确定潜在的Tspan18结合伙伴，来表征Tspan18在神经嵴发育中的作用。我提出这一建议的基本原理是，确定Tspan18在神经嵴中的作用将使我们深入了解协调早期神经嵴发育的复杂信号网络，并可能为理解神经母细胞瘤等疾病的进展提供线索。公共卫生相关性：神经嵴细胞是在具有骨干的胚胎中形成不同结构的迁移干细胞。当神经嵴细胞调节出错时，就会导致各种出生缺陷和癌症。这项研究将描述一种新的蛋白tetraspanin18，初步证据表明它是神经嵴发育所必需的。

DESCRIPTION (provided by applicant): The neural crest is a unique, migratory population of stem cells in vertebrate embryos that gives rise to numerous and varied tissues including the peripheral nervous system, craniofacial skeleton, cardiac outflow tract, and melanocytes. The vast number of disorders caused by deregulation of neural crest development, including birth defects such as orofacial clefts and cancers such as neuroblastoma, reflect the diversity of neural crest derivatives. In spite of the key role played by neural crest cells in development and disease, the mechanisms that control the formation of migratory neural crest cells are poorly understood. My preliminary data identify tetraspanin18 (Tspan18) as a novel, critical regulator of neural crest migratory properties. Tetraspanins are transmembrane proteins that organize and regulate cell signaling, motility, and adhesion. The objective of my research is to characterize the role of Tspan18 in neural crest development. I hypothesize that Tspan18 is part of an essential scaffold responsible for organizing signals at the neural crest cell membrane through interactions with other proteins, and that Tspan18 is required for proper neural crest development. Specifically, the proposed research will characterize the role of Tspan18 in neural crest development by (1) determining the requirement for Tspan18 in the neural crest through antisense morpholino oligonucleotide loss-of-function assays, and (2) identifying potential Tspan18 binding partners by affinity purification and mass spectrometry. My rationale for this proposal is that defining the role of Tspan18 in the neural crest will give us insight into the complex web of signals that coordinates early neural crest development, and may provide clues to understanding the progression of disease, such as neuroblastoma. PUBLIC HEALTH RELEVANCE: Neural crest cells are migrating stem cells that form diverse structures in embryos with backbones. When neural crest cell regulation goes awry, a variety of birth defects and cancers results. This research will characterize a novel protein tetraspanin18 that preliminary evidence suggests is required for neural crest development.