K99 to R00 Transition-Neural Crest Migration and Survival in Zebrafish

其应用的目的是以斑马鱼为节点，解剖Foxd3介导的神经脊(NC)细胞存活和迁移末端的分子连接，以确定这些途径是否促进体内NG来源的肿瘤的转移。斑马鱼因其与人类基因组的紧密同步性以及调节组织和器官发育的保守的分子途径，为进行此类研究提供了一个强大的工具。我们先前鉴定并鉴定了一个斑马鱼foxd3突变系，该突变系具有特定的NC细胞存活和迁移缺陷。我们还发现Snai1b转录因子是Foxd3功能的关键调节因子。这一应用的基本假设是，对Foxd3通路的了解将为NC来源的肿瘤的转移提供一个理解的环境和要求。在目标1中，遗传上位性和生物化学将决定Foxd3是否直接激活Snai1b的表达。这一目标也将决定SNAI 1b是否通过抑制一个或多个BH3-Only基因的表达来抑制NC的凋亡，并通过抑制钙粘素-6的表达来促进NC的迁移。在目标2中，将分析在bK99阶段期间在微阵列筛选中识别的基因的列表，以确定其NC表达模式和敲除表型，以及它们的能力 挽救foxd3突变体和SNAI 1b MO注射胚胎中的NC缺陷。在目标3中，foxd3和Snai1b将在发育中的NC细胞中表达，以研究它们在促进已建立的斑马鱼NC肿瘤转移中的潜在作用，包括在K99期发现的新模型。 罗德尼·斯图尔特将继续接受遗传学、神经生物学和肿瘤学方面的高级培训 在他的职业生涯中。这项研究提案旨在支持候选人在犹他州大学亨斯迈癌症研究所肿瘤学系任教的头3年内作为独立调查员。 犹他州州立大学亨茨南癌症研究所

The goal oflhis application is to use the zebrafish as a |nodel to dissect the molecular connponents of Foxd3-mediated neural crest (NC) cell survival and migration end to determine if these pathways promote metastasis in NG-derived tumors in vivo. The zebrafish with its close synteny to the human genome and its conserved molecular pathways regulating the development of tissues and organs, Offers a powerful tool with which to conduct such research. We previoulsy identified and characterized a zebrafish foxd3 mutant line that has specific NC ceil survival and migration defects. We also identified the Snai1b transcription factor-as a critical mediator of Foxd3 function. The underlying hypothesis of this applicatipn is that knowledge of the Foxd3 pathway wiII provide an understanding environments, requirement for the metaststasis of NC-derived tumors. In Aim 1, genetic epistasis and biochemistry will determine if Foxd3 directly activates snai1b expression. This aim will also will also determine whether Snai 1b inhibits NC apoptosis by repressing the expression of one or more BH3-only genes, and promotes NC migration by repressing cadherin-6 expression . In Aim 2, a list of genes identified in a microarray screen during the bK99 phase will abe analyzed for thier NC expression pattern and knockdown phenotypes, as well as their ability to rescue NC defects in foxd3 mutant and snai 1b MO-injected embryos. In Aim 3, foxd3 and snai1b will be expressed in developing NC cells to examine their potential roles in promoting metastasis in established zebrafish NC tumors, including a new model identified during the K99 phase. Rodney Stewart will continue to receive advanced training in genetics, neurobiology and oncology throughout his career. This research proposal is designed to support the candidate as an independent investigator during the first 3 years of a faculty appointment in the Department of Oncology Sciences at the Huntsman Cancer Institute at the University of Utah. Huntsrnan Cancer Institute at the yniyersity of-Utahnd