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.

项目摘要 颅面畸形伴随着三分之一的出生缺陷，孤立或非综合征唇裂， 全世界1/700的新生儿中仅发生腭部（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。环境因素，如叶酸摄入量和吸烟， 然而，母体遗传效应难以建模和研究。 这项建议的目的是研究有史以来第一次保护性的母亲对腭发育的遗传效应。到 据我们所知，在小鼠模型中没有描述过任何分娩的保护性母体遗传效应。 缺损我们的数据显示Specc 11 DCCD 2/+和Thm 1aln/+单杂合子导致~20%（n=45）和0%（n = 25）的突变。 （n=24）CP组。而Specc 11 DCCD 2/+; Thm 1aln/+双杂合子的CP值为33%（n=30）。 然而，只有当与Specc 1 lDCCD 2/+母体进行杂交时，才观察到CP的发生。 与Thm 1aln/+母体杂交，单杂合子（n=20）和双杂合子的CP均为0 （n=25）。由于Specc 11 DCCD 2/+雄性与野生型雌性杂交，Specc 11 DCCD 2/+中仍产生约20%的CP 杂合子（n=20），我们排除了Specc 11 DCCD 2/+母亲的负作用或Thm 1aln/+的保护作用 父亲.因此，我们假设，Thm 1aln/+女性提供了一个保护性的母性遗传效应CP。 我们将通过研究Aim 1的母体环境和确定Aim 1的分子结构来验证我们的假设。 Aim 2中保护作用的性质。母体环境将通过胚胎移植进行评估 实验和子宫特异性Thm 1杂合性的产生。保护剂的分子性质 将通过评估母体和哺乳动物中的表观遗传、转录组和蛋白质组学变化来确定效果。 胚胎组织Thm 1和Specc 1 l缺陷均影响纤毛发生。因此，我们的细胞和分子 研究将集中在保护作用背后的细胞骨架和纤毛信号变化。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验证的假设 单独的CP复合体疾病