Investigating the protective effect of maternal Thm1 heterozygosity against cleft palate

项目总结 所有出生缺陷中有三分之一伴有头面部畸形，伴有孤立或非综合征性唇裂和唇裂。 仅在全世界出生的婴儿中就有1/700人患有腭裂(CL/P)。这些孤立的CL/P具有复杂的病因，包括 既有遗传因素也有环境因素。环境因素，如叶酸摄入量和吸烟，一直是 然而，母体遗传效应被证明会影响母体环境，因此很难建立模型并进行研究。 这项建议的目的是研究有史以来第一次保护性母体遗传对腭裂发生的影响。至 据我们所知，在小鼠模型中，保护性母体遗传效应尚未被描述为任何出生 叛逃。我们的数据显示，specc1lDCCD2/+和Thm1aln/+单一杂合子导致约20%(n=45)和0% CP组24例。而Spec1lDCCD2/+、Thm1aln/+双重杂合子的CP为33%(n=30)。 然而，只有当与specc1lDCCD2/+母亲进行杂交时，才能观察到这种CP的发生。 在Thm1aln/+母亲中，相同的杂交导致单一(n=20)和双重杂合子的CP为0% 25例。因为，specc1lDCCD2/+雄性与野生型雌性杂交仍会导致specc1lDCCD2/+的CP为20% 杂合子(n=20)，我们排除了specc1lDCCD2/+母亲的负面影响或Thm1aln/+的保护作用 父亲。因此，我们假设Thm1aln/+女性为CP提供了一种保护性母体遗传效应。 我们将通过调查Aim1的母体环境和确定分子水平来验证我们的假设 AIM2的保护作用的性质。母体环境将通过胚胎移植进行评估。 子宫特异性Thm1杂合子的实验和生成。保护性的分子本质 影响将通过评估表观遗传学、转录本和蛋白质组的变化来确定 胚胎组织。Thm1和Spec1缺乏症均影响纤毛发生。因此，我们的细胞和分子 研究的重点将是细胞骨架和纤毛信号的变化，这些变化是保护作用的基础。这些研究 将产生关于母体环境在病因学中的作用的新的见解和可检验的假设 分离的CP复合体疾病。

PROJECT SUMMARY Craniofacial anomalies accompany a third of all birth defects, with isolated or nonsyndromic clefts of the lip and palate (CL/P) alone occurring in 1/700 births worldwide. These isolated CL/P have a complex etiology including both genetic and environmental factors. Environmental factors such as folate intake and smoking have been shown to affect maternal environment, however, maternal genetic effects have been difficult to model and study. The objective of this proposal is to study the first-ever protective maternal genetic effect on palatogenesis. To our knowledge, a protective maternal genetic effect in a mouse model has not been described for any birth defect. Our data show that Specc1lDCCD2/+ and Thm1aln/+ single heterozygotes resulted in ~20% (n=45) and 0% (n=24) CP respectively. In contrast, Specc1lDCCD2/+;Thm1aln/+ double heterozygotes showed ~33% CP (n=30). However, this occurrence of CP was observed only when the cross was performed with Specc1lDCCD2/+ mothers. With Thm1aln/+ mothers, the same cross resulted in 0% CP in both single (n=20) and double heterozygotes (n=25). Since, a Specc1lDCCD2/+ male crossed with wildtype female still resulted in ~20% CP in Specc1lDCCD2/+ heterozygotes (n=20), we ruled out a negative effect by Specc1lDCCD2/+ mothers or protective effect by Thm1aln/+ fathers. Thus, we hypothesized that the Thm1aln/+ female provides a protective maternal genetic effect for CP. We will test our hypothesis by investigating the maternal environment in Aim1 and by determining the molecular nature of the protective effect in Aim2. The maternal environment will be evaluated by embryo transfer experiments and generation of uterine-specific Thm1 heterozygosity. The molecular nature of the protective effect will be determined by assessing epigenetic, trancriptomic, and proteomic changes in maternal and embryonic tissue. Both Thm1 and Specc1l deficiency affects ciliogenesis. Thus, our cellular and molecular studies will focus on cytoskeletal and ciliary signaling changes underlying the protective effect. These studies will generate novel insights and testable hypotheses regarding the role of maternal environment in the etiology of the isolated CP complex disease.