Genetic Regulation of Valproic Acid Teratogenicity

描述（由申请人提供）： 宫内暴露于药物化合物Depakene（丙戊酸; VPA）会增加人类先天性畸形和神经发育缺陷的发生率。本研究的目的是鉴定和表征VPA诱导的神经管缺陷（NTDs）的遗传修饰剂。拟议的研究是在小鼠模型中进行的，但所得到的数据预计将使临床医生在FDA批准下，根据人类患者群体中同源基因变异带来的风险开发个性化的药物策略。这一点越来越重要，因为VPA现在不仅用于癫痫治疗，还用于治疗偏头痛、双相情感障碍和双相情感障碍，增加了育龄妇女的暴露。我们假设NTD和神经发育缺陷的病因涉及胎儿遗传贡献，并且其蛋白质产物正在改变VPA诱导神经异常的能力。具体而言，我们建议测试的假设，酰基辅酶A合成酶中链（ACSM）基因内的变化是更普遍的NTD小鼠胚胎暴露于致畸浓度的VPA后，比在类似暴露的胚胎，似乎正常。我们建议使用我们建立的VPA致畸小鼠模型，对7号染色体上的关键区域进行精细定位，以确定负责VPA诱导的NTDs和神经发育缺陷的新基因变体。我们建议重新测序所有已知的基因映射到关键区域在受影响和未受影响的小鼠胎儿在子宫内VPA暴露。将使用样本的靶向富集（NimbleGen序列捕获阵列和Agilent Technologies SureSelect系统），通过下一代测序技术结合使用桑格测序和靶向重测序，彻底询问VPA敏感性基因座。在下一代测序之前进行靶点富集的目标是扩展我们的全基因组关联研究，以：1）定义SNP和独特的基因组变异; 2）将这些变异与VPA诱导的神经发育疾病相关联。然后，我们将使用现有的靶向胚胎干细胞克隆开发新的ACSM突变小鼠品系，并验证该基因家族对VPA诱导的NTD的易感性的调节作用。

DESCRIPTION (provided by applicant): The in utero exposure to the pharmaceutical compound Depakene (Valproic Acid; VPA) increases the incidence of congenital malformations and neurodevelopmental defects in humans. The objective of this research proposal is to identify and characterize the genetic modifiers of VPA-induced neural tube defects (NTDs). The proposed research is conducted in mouse models, but the resultant data is expected to enable clinicians, under FDA approval, to develop personalized medicine strategies based on an the risk posed by homologous gene variants in human patient populations. This is increasingly important as VPA is now used not only in epilepsy therapy, but also to treat migraine headaches, and bipolar and schizoaffective disorders, increasing exposure to women of reproductive age. We hypothesize that the etiology of NTDs and neurodevelopmental deficits involves fetal genetic contributions, and that their protein products are modifying the ability of VPA to induce neural abnormalities. Specifically, we propose to test the hypothesis that variation within Acyl-CoA Synthetase Medium-Chain (ACSM) genes are more prevalent in mouse embryos with NTDs following exposure to teratogenic concentrations of VPA, than in similarly exposed embryos that appear normal. We propose to fine map the critical region on chromosome 7 in order to identify novel gene variants responsible for NTDs and neurodevelopmental deficits induced by VPA, using our established mouse models of VPA-teratogenesis. We propose to re-sequence all known genes mapped to the critical region in affected and unaffected mouse fetuses following in utero VPA exposure. A combination of Sanger sequencing and targeted resequencing by next-generation sequencing techniques, using targeted enrichment (NimbleGen Sequence Capture Arrays and Agilent Technologies SureSelect System) of samples will be used to thoroughly interrogate the VPA-sensitivity loci. The goal of target enrichment prior to next-generation sequencing is to expand upon our whole genome association studies to: 1) define SNPs and unique genomic variations; and 2) correlate these variants to VPA-induced neurodevelopmental disease. We will then develop novel ACSM mutant mouse lines using existing targeted embryonic stem cell clones and validate this gene family as regulating susceptibility to VPA-induced NTDs.