Role of the Foxe3 Gene Family in Lens Formation.

Foxe3 基因家族在晶状体形成中的作用。

• 批准号: 8548460
• 负责人: MILAN Alexander JAMRICH
• 金额: $ 23.67万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8548460
• 关键词: Affect; Anterior; Cells; Defect; Development; Developmental Biology; Developmental Gene; Developmental Process; Diagnosis; Diagnostic Procedure; Ectoderm; Epithelium; Event; Eye Development; Eye diseases; Gene Expression Profile; Gene Family; Gene Targeting; Genes; Goals; Head; Human; Knowledge; Lead; Lens Fiber; Lens Placodes; Maintenance; Mammals; Mediating; Mediator of activation protein; Molecular; Molecular Biology; Morphology; Mus; Mutant Strains Mice; Mutate; Mutation; Neuroectoderm; Optic vesicle; Physiology; Play; RNA Sequences; Research; Retinal; Role; Staging; Surface Ectoderm; Techniques; Undifferentiated; Vesicle; Vision; chromatin immunoprecipitation; fiber cell; gene therapy; in utero; lens; mutant; novel diagnostics; transcription factor

DESCRIPTION (provided by applicant): During early vertebrate development, lens is induced to form from the head surface ectoderm by the presumptive retinal neuroectoderm. When the evaginating optic vesicle contacts the surface ectoderm, the ectoderm thickens and forms a lens placode. In mammals, the lens placode invaginates and forms the lens vesicle. The posterior lens cells stop proliferation and differentiate into lens fiber cells. The anterior lens cells form a relatively undifferentiated, proliferatively active lens epithelium. Our long-term goal is to identify developmental steps and molecular events necessary for lens formation. The primary focus of this research is the role and the mechanism of action of the forkhead gene Foxe3. This research is a natural continuation of our previous studies investigating the role of this gene in lens formation. Since normal development and maintenance of lens cells is critical for vision, it is essential to define the developmental processes and gene networks that govern the development and survival of lens cells. Several key genes control of the formation, proliferation and differentiation of lens cells. One of them is the forkhead gene Foxe3. This gene encodes a transcription factor and mutations in this gene cause abnormal lens development in mouse and human. The absence of Foxe3 function leads to changes in the proliferation, differentiation and survival of lens cells. How the loss of Foxe3 leads to all of these changes remains largely unknown. Since Foxe3 is a transcription factor, its effects on the physiology and morphology of lens cells has to be mediated by other genes. With few exceptions, the downstream mediators of Foxe3 are not known. To better understand how Foxe3 regulates lens development and why mutations in this gene lead to such dramatically abnormal lens development, we will in Specific Aim 1 identify genes that belong to the Foxe3 lens regulatory network by comparing transcriptomes of wild type and Foxe3-deficient lenses using deep RNA-sequencing. In Specific Aim 2, we will identify direct target genes of Foxe3 by chromatin immunoprecipitation combined with sequencing. These cutting edge techniques in molecular and developmental biology will help us identify genes that mediate Foxe3 function during lens development. In Specific Aim 3, we will correct the molecular and phenotypic lens defects in mice with mutant or absent Foxe3 using in utero gene therapy. This research will lead to a better understanding of lens development, as this gene is regulating the early critical steps in lens formation. However, since mutations in this gene cause abnormal eye development in mouse and human, this research will result in knowledge that will allow a better diagnosis and treatment of diseases of the eye in which the components of the Foxe3 regulatory network are mutated.

描述(申请人提供)：在脊椎动物早期发育期间，晶状体是由假定的视网膜神经外胚层从头部表面的外胚层诱导形成的。当外溢的视泡接触表面外胚层时，外胚层增厚并形成晶状体胎座。在哺乳动物中，晶状体胎盘内陷并形成晶状体小泡。后方晶状体细胞停止增殖，分化为晶状体纤维细胞。晶状体前细胞形成相对未分化的、增殖活跃的晶状体上皮。我们的长期目标是确定晶状体形成所需的发育步骤和分子事件。本研究的重点是叉头基因Foxe3的作用和作用机制。这项研究是我们之前研究该基因在晶状体形成中的作用的自然延续。由于晶状体细胞的正常发育和维持对视力至关重要，因此确定支配晶状体细胞发育和存活的发育过程和基因网络是至关重要的。控制晶状体细胞形成、增殖和分化的几个关键基因。其中之一是叉头基因foxe3。该基因编码一种转录因子，该基因的突变会导致小鼠和人类晶状体发育异常。Foxe3功能缺失会导致晶状体细胞增殖、分化和存活的改变。失去Foxe3是如何导致所有这些变化的，目前在很大程度上尚不清楚。由于Foxe3是一种转录因子，它对晶状体细胞的生理和形态的影响必须通过其他基因来调节。除了少数例外，Foxe3的下游介体是未知的。为了更好地了解Foxe3是如何调控晶状体发育的，以及为什么该基因的突变会导致晶状体发育如此显著的异常，我们将针对特定的目标1，通过比较野生型和Foxe3缺陷晶状体的转录产物，使用深度RNA测序来鉴定属于Foxe3晶状体调控网络的基因。在特定目标2中，我们将通过染色质免疫沉淀和测序相结合的方法鉴定Foxe3的直接靶基因。这些分子和发育生物学的尖端技术将帮助我们识别在晶状体发育过程中调节Foxe3功能的基因。在特定的目标3中，我们将使用宫内基因治疗来纠正Foxe3突变或缺失的小鼠的分子和表型晶状体缺陷。这项研究将有助于更好地理解晶状体的发育，因为这个基因正在调节晶状体形成的早期关键步骤。然而，由于该基因的突变会导致小鼠和人类眼睛发育异常，这项研究将带来更好的眼部疾病诊断和治疗知识，在这些疾病中，Foxe3调控网络的组成部分发生突变。