Deciphering the role of mediator of ErbB2 driven cell motility (Memo1) in regulat

破译 ErbB2 驱动的细胞运动介质 (Memo1) 在调节中的作用

• 批准号: 8718389
• 负责人: Eric Van Otterloo
• 金额: $ 5.51万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-05 至 2017-05-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8718389
• 关键词: Address; Affect; Alleles; Automobile Driving; Biological Models; Birth; Cancer Model; Cell Culture Techniques; Cell Proliferation; Cell physiology; Cells; Cellular biology; Cephalic; Cessation of life; Congenital Abnormality; Cues; Defect; Detection; Development; Disease; Economic Burden; Ectoderm; Elements; Embryo; Embryonic Development; Ethylnitrosourea; Event; Face; Ganglia; Genes; Genetic; Genetic Models; Goals; Growth; Head; Human; In Vitro; Incidence; Knowledge; Link; Live Birth; Mediating; Mediator of activation protein; Mesenchyme; Migration Assay; Modeling; Mus; Mutagenesis; Mutation; Neural Crest; Neural Crest Cell; Palate; Pattern; Pattern Formation; Phenotype; Play; Prevention; Process; Proteins; Quality of life; Regulation; Robin bird; Role; Secondary Palate; Series; Signal Transduction; Skeleton; Structure; Structure-Activity Relationship; System; Tissues; Transgenes; Work; base; cell behavior; cell motility; comparative; craniofacial; craniofacial complex; cranium; developmental disease; embryonic stem cell; extracellular; insight; meningioma; migration; mutant; novel; null mutation; orofacial; population based; prevent; public health relevance; research study; response; skeletal; skull base; stem cell population

DESCRIPTION (provided by applicant): Embryonic development of craniofacial structures requires a tightly orchestrated series of cellular and morphological events. These include regulation of cellular proliferation, growth, migration, and differentiation. The sensitivity of thse processes to perturbation is evident by the high incidence of human birth defects affecting the head and associated structures (~75% of all birth defects have a craniofacial component), most common of which is orofacial clefting (~1:600-1,000 live births). These defects will impart a significant decrease in quality of life on those afflicted and present a major economic burden associated with treatment. To better understand and ultimately prevent these disorders prenatally a precise understanding of genes controlling the aforementioned cellular processes during normal craniofacial development is required. To this end, through an ENU based mutagenesis screen, we have identified Mediator of ErbB2-driven cell motility 1 (Memo1) as a novel regulator of multiple aspects of craniofacial development, including appropriate formation of the skull-base and palate. Interestingly, most of the affected structures in Memo1enu/enu mutants are derived from the cranial neural crest cells, implicating a role for Memo1 within this embryonic stem-cell population. Previous studies identified Memo1 as an important component in modulating extracellular cues into intracellular responses, such as cellular migration and proliferation. However, Memo1-null embryos are early embryonic lethal, precluding any analysis of Memo1's role during craniofacial development. Our Memo1 ENU-allele thus provides a unique model to decipher Memo1's role during these processes. Our general goal is to precisely characterize the mechanistic role of Memo1 during craniofacial development using two Aims. (AIM 1) First, by combining our Memo1 ENU-allele with a Memo1 null-allele, we will generate an allelic series, serving as a convenient genetic model system to dissect Memo1's cellular role during craniofacial development. The resultant allelic combinations will be utilized for analysis o gross craniofacial development and cranial neural crest cell biology, including a more detailed in vitro examination of neural crest cell migration. (AIM 2) Secondly, because Memo1 is expressed in multiple craniofacial tissues, we will generate a conditional allele of Memo1 allowing its tissu specific deletion. Given the majority of the defects in our ENU- mutant are found in structures derived from the cranial neural crest cells, we will specifically assess the cell autonomous requirements of Memo1 in this tissue by generating mutants with a neural crest specific deletion of Memo1, and thoroughly characterize how this impacts craniofacial development. In summary, Memo1 has only recently been linked with face formation but further characterization of this gene is likely to generate insight into the broader gene network responsible for normal human facial formation. In the long-term, the work proposed will contribute to a more detailed understanding of processes involved in craniofacial development, providing a clearer path towards detection and prevention of craniofacial disorders.

描述（由申请人提供）：颅面结构的胚胎发育需要一系列紧密协调的细胞和形态学事件。这些包括细胞增殖、生长、迁移和分化的调节。这些过程对扰动的敏感性通过影响头部和相关结构的人类出生缺陷的高发生率（约75%的所有出生缺陷具有颅面成分）而明显，其中最常见的是口面裂（约1：600- 1，000活产）。这些缺陷将使患者的生活质量显著下降，并带来与治疗相关的重大经济负担。为了更好地了解并最终预防这些疾病产前基因控制上述细胞过程中正常颅面发育的精确理解是必要的。为此，通过基于ENU的诱变筛选，我们已经确定ErbB 2驱动的细胞运动1（Memo 1）的调解人作为颅面发育的多个方面，包括颅底和腭的适当形成的一种新的调节剂。有趣的是，Memo 1 enu/enu突变体中大多数受影响的结构来自颅神经嵴细胞，这暗示了Memo 1在胚胎干细胞群中的作用。以前的研究确定Memo 1作为一个重要的组成部分，在调节细胞外的线索到细胞内的反应，如细胞迁移和增殖。然而，Memo 1基因缺失的胚胎是早期胚胎致死的，排除了Memo 1在颅面发育过程中的作用的任何分析。因此，我们的Memo 1 ENU等位基因提供了一个独特的模型来破译Memo 1在这些过程中的作用。我们的总体目标是精确地描述Memo 1在颅面发育过程中的机械作用，使用两个目标。(AIM 1）首先，通过将我们的Memo 1 ENU等位基因与Memo 1空等位基因相结合，我们将产生一个等位基因系列，作为一个方便的遗传模型系统来剖析Memo 1在颅面发育过程中的细胞作用。所得等位基因组合将用于大体颅面发育和颅神经嵴细胞生物学分析，包括神经嵴细胞迁移的更详细体外检查。(AIM 2）其次，由于Memo 1在多种颅面组织中表达，我们将产生Memo 1的条件等位基因，允许其组织特异性缺失。鉴于我们的ENU突变体中的大多数缺陷存在于源自颅神经嵴细胞的结构中，我们将通过产生具有Memo 1的神经嵴特异性缺失的突变体来具体评估Memo 1在该组织中的细胞自主需求，并彻底表征这如何影响颅面发育。总之，Memo 1只是最近才与面部形成有关，但对该基因的进一步表征可能会深入了解负责正常人类面部形成的更广泛的基因网络。从长远来看，拟议的工作将有助于更详细地了解颅面发育过程，为颅面疾病的检测和预防提供更清晰的途径。