Investigating the protective effect of maternal Thm1 heterozygosity against cleft palate

母体 Thm1 杂合性对腭裂的保护作用研究

• 批准号: 10742414
• 负责人: Irfan Saadi
• 金额: $ 23.25万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10742414
• 关键词: Actins; Affect; Alleles; Birth; Cells; Cilia; Circulation; Cleft Palate; Coiled-Coil Domain; Complex; Congenital Abnormality; Congenital omphalocele; Craniofacial Abnormalities; Cytoskeletal Proteins; Cytoskeleton; Data; Disease; Embryo; Embryo Transfer; Environment; Environmental Risk Factor; Epigenetic Process; Erinaceidae; Etiology; Exencephalies; Fathers; Female; Folic Acid; Future; GLI gene; Generations; Genes; Genetic; Genotype; Goals; Heterozygote; Human; Incidence; Intake; Mediating; Methods; Microtubules; Modeling; Molecular; Monitor; Mothers; Mus; Mutant Strains Mice; Mutation; Nature; Orbital separation excessive; Outcome; Parents; Phenotype; Placenta; Proteomics; Role; Signal Pathway; Signal Transduction; Smoking; Syndrome; Testing; Therapeutic; Tissues; Uterus; autosome; calponin; cilium biogenesis; cleft lip and palate; embryo tissue; experimental study; fetal; gain of function; histone methylation; improved; insight; interest; male; mouse model; mutant; next generation sequencing; novel; orofacial cleft; palatal shelves; palatogenesis; promoter; protective effect; retrograde transport; transcriptome sequencing

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.

项目概要 三分之一的出生缺陷伴有颅面异常，其中包括孤立性或非综合征性唇裂和唇裂。 全球 1/700 的新生儿中仅存在上颚 (CL/P)。这些孤立的 CL/P 具有复杂的病因，包括 遗传和环境因素都有。叶酸摄入量和吸烟等环境因素 然而，母体遗传效应已被证明会影响母体环境，但很难对其进行建模和研究。 该提案的目的是研究有史以来第一个对腭发育的保护性母体遗传效应。到 据我们所知，小鼠模型中的保护性母体遗传效应尚未被描述为任何出生 缺点。我们的数据显示 Specc1lDCCD2/+ 和 Thm1aln/+ 单杂合子导致约 20% (n=45) 和 0% (n=24)分别为CP。相比之下，Specc1lDCCD2/+；Thm1aln/+ 双杂合子显示~33% CP (n=30)。 然而，仅当与 Specc11DCCD2/+ 母本杂交时才观察到这种 CP 的发生。 对于 Thm1aln/+ 母亲，相同的杂交导致单杂合子 (n=20) 和双杂合子的 CP 均为 0% （n=25）。因为，Specc1lDCCD2/+ 雄性与野​​生型雌性杂交仍然导致 Specc1lDCCD2/+ 中约 20% 的 CP 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 父亲们。因此，我们假设 Thm1aln/+ 雌性为 CP 提供保护性母体遗传效应。 我们将通过调查 Aim1 中的母体环境并确定分子水平来检验我们的假设。 Aim2 中保护作用的性质。将通过胚胎移植评估母体环境 子宫特异性 Thm1 杂合性的实验和生成。保护剂的分子性质 效果将通过评估母体和母体内的表观遗传、转录组和蛋白质组变化来确定。 胚胎组织。 Thm1 和 Specc1l 缺陷都会影响纤毛发生。因此，我们的细胞和分子 研究将重点关注保护作用背后的细胞骨架和纤毛信号传导变化。这些研究 将产生关于母体环境在病因学中的作用的新见解和可检验的假设 孤立的 CP 复杂疾病。