GENE DISCOVERY FOR CRANIOFACIAL DISORDERS

描述(申请人提供)：出生缺陷影响大约5%的美国婴儿，四分之三涉及头部、面部和口腔组织。最常见的头面部出生缺陷是口面部裂：在美国，唇裂+/-腭裂(CL/P)在每1100名新生儿中约有1例，而腭裂在每1600名新生儿中约有1例。因此，口裂是一个相当大的公共卫生问题和费用，而且大多数病例现在都无法预测或预防。孤立的“非综合征”CL/P(nsCL/P)是最常见的头面部出生缺陷，约占所有CL/P病例的70%。nsCL/P是一种非孟德尔式的多基因疾病，由于多个基因，每个基因发挥的作用相对较小，彼此之间和环境因素相互作用，最终导致特定通路和遗传网络在胎儿颅面发育过程中的缺陷作用。然而，目前对影响nsCL/P的基因很少，也没有任何环境影响是确定的。该方案的目的是确定与颅面发育有关的基因、途径和遗传网络，这些基因、途径和遗传网络代表了nsCL/P的遗传和非遗传决定因素的潜在靶点。识别这些靶点对于设计最终旨在预防这种衰弱和毁容的出生缺陷的治疗策略是必要的。目前，准确研究人类头面部发育过程中的基因活动是非常困难的。因此，我们计划对小鼠发育期面部的基因表达谱进行仔细的微阵列研究，其中遗传背景(C57BL/6J)、仔细的胎龄选择、多个时间点的采样以及对许多重复样本的分析都可以很容易地实现。此外，我们计划在C57BL/6J背景下，对Ski基因缺失敲除等位基因纯合的小鼠面部发育进行类似的研究。这些Ski-/-小鼠具有极高的正中线面部裂纹率，为异常颅面发育过程中的基因表达提供了宝贵的比较，特别是应该识别对Ski反应的发育相的遗传路径和网络。对于在面部发育过程中显得特别重要的基因，我们将识别和验证人类SNPs，用于未来nsCL/P的连锁和关联研究。我们将应用最先进的生物信息学工具来分析和解释数据，所有这些数据都将存放在适当的公共数据库中。

DESCRIPTION (provided by applicant): Birth defects affect approximately 5% of all infants in the USA, three-fourths involving the head, face, and oral tissues. The most frequent craniofacial birth defects are orofacial clefts: cleft lip +/- cleft palate (CL/P) affects ~1 per 1100 births in the USA, and cleft palate ~1 per 1600 births. Orofacial clefts thus represent a considerable public health problem and expense, and most cases cannot now be predicted or prevented. Isolated, "non-syndromic" CL/P (nsCL/P) is the most common craniofacial birth defect, accounting for approximately 70% of all cases of CL/P. nsCL/P is a non-Mendelian, multifactorial disorder, due to multiple genes, each exerting a relatively small effect, interacting with each other and with environmental factors to ultimately result in defective action of specific pathways and genetic networks during fetal craniofacial development. However, few of the genes, and none of the environmental influences, that contribute to nsCL/P are currently known with certainty. The goal of this proposal is to identify the genes, pathways, and genetic networks that are involved in craniofacial development and that thus represent potential targets for genetic and non-genetic determinants of nsCL/P. Identification of these targets will be necessary to devise therapeutic strategies ultimately aimed at preventing this debilitating and disfiguring birth defect. It is currently very difficult to accurately study gene action during craniofacial development in the human. Accordingly, we plan a careful microarray study of gene expression profiles in the developing face of the mouse, in which genetic background (C57BL/6J), careful timing of fetal age, sampling at numerous timepoints, and analyses of many replicate samples can all be readily achieved. In addition, we plan an analogous study of facial development in mice homozygous for a null knockout allele of the Ski locus, carried on the C57BL/6J background. These Ski -/- mice have an exceedingly high rate of midline facial clefts, providing an invaluable comparison of gene expression during aberrant craniofacial development that, in particular, should identify genetic pathways and networks of the developing facies that are responsive to Ski. For genes that appear of particular importance during facial development, we will identify and validate human SNPs for use in future linkage and association studies of nsCL/P. We will apply state-of-art bioinformatics tools to analyze and interpret the data, all of which we will deposit in appropriate public data repositories.