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

这个应用程序的目标是将斑马鱼作为一种|我们需要剖析Foxd 3介导的神经嵴（NC）细胞存活和迁移结束的分子组成，以确定这些途径是否促进体内NG衍生肿瘤的转移。斑马鱼与人类基因组的同线性关系密切，其保守的分子途径调节组织和器官的发育，提供了一个强大的工具，进行这样的研究。我们先前鉴定并表征了具有特定NC细胞存活和迁移缺陷的斑马鱼foxd 3突变系。我们还确定了Snai 1b转录因子作为Foxd 3功能的关键介质。本申请的基本假设是Foxd 3途径的知识将提供对NC衍生的肿瘤的抑制的理解环境和要求。在目标1中，遗传上位性和生物化学将确定Foxd 3是否直接激活snai 1b表达。这一目标也将确定Snai 1b是否通过抑制一个或多个仅BH 3基因的表达来抑制NC凋亡，并通过抑制钙粘蛋白-6表达来促进NC迁移。在目的2中，将分析在bK 99期期间在微阵列筛选中鉴定的一系列基因的NC表达模式和敲低表型，以及它们在bK 99期表达的能力。 拯救foxd 3突变体和snai 1b MO注射胚胎中的NC缺陷。在目标3中，foxd 3和snai 1b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期鉴定的新模型。 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。本研究提案旨在支持候选人在犹他州大学亨茨曼癌症研究所肿瘤科学系担任教职的前3年内担任独立研究者。 犹他州亨茨南癌症研究所

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