Characterizing the Role of Tspan18 in Neural Crest Development

描述(申请人提供)：神经脊是脊椎动物胚胎中干细胞的一种独特的、可迁移的群体，它产生了大量和不同的组织，包括周围神经系统、颅面部骨骼、心脏流出道和黑素细胞。由于放松对神经脊发育的管制而导致的大量疾病，包括先天缺陷，如口裂和癌症，如神经母细胞瘤，反映了神经脊衍生品的多样性。尽管神经脊细胞在发育和疾病中起着关键作用，但控制迁移性神经脊细胞形成的机制却知之甚少。我的初步数据表明，Tspan18(Tspan18)是神经脊游走特性的一种新的、关键的调节因子。Tetraspanins是一种跨膜蛋白，可以组织和调节细胞的信号、运动和黏附。我研究的目的是确定Tspan18在神经脊发育中的作用。我假设Tspan18是负责通过与其他蛋白质相互作用在神经脊细胞膜上组织信号的必要支架的一部分，并且Tspan18是正常神经脊发育所必需的。具体地说，这项研究将通过(1)通过反义吗啉寡核苷酸功能丧失分析来确定神经脊中Tspan18的需求，以及(2)通过亲和纯化和质谱分析来确定潜在的Tspan18结合伙伴，从而表征Tspan18在神经脊发育中的作用。我提出这一建议的理由是，定义Tspan18在神经脊中的作用将使我们深入了解协调早期神经脊发育的复杂信号网络，并可能为理解疾病的进展提供线索，如神经母细胞瘤。与公共健康相关：神经脊细胞正在迁移干细胞，这些干细胞在带有脊椎的胚胎中形成不同的结构。当神经脊细胞的调节出现偏差时，就会导致各种出生缺陷和癌症。这项研究将表征一种新的蛋白质Tetraspanin18，初步证据表明，这种蛋白质是神经脊发育所必需的。

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.