In utero rescue of cleft palate using maternal administration of folic acid

使用叶酸在子宫内挽救腭裂

• 批准号: 10646021
• 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: 2023-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10646021
• 关键词: Actins; Actomyosin; Affect; Alleles; Binding; Birth; Cell Proliferation; Cells; Cleft Palate; Coiled-Coil Domain; Complex; Congenital Abnormality; Craniofacial Abnormalities; Cytoskeleton; Data; Defect; Development; Dose; Early Intervention; Embryo; Environment; Environmental Risk Factor; Etiology; F-Actin; Female; Folic Acid; Future; Genetic; Goals; Growth; Hour; Human; Incidence; Intake; Intervention; Intervention Studies; Intraperitoneal Injections; Knock-in; Knowledge; Mediating; Mesenchyme; Methodology; Methods; Microtubules; Modeling; Molecular; Molecular Target; Mus; Mutant Strains Mice; Mutation; Myosin Type II; Neural Tube Closure; Neural Tube Defects; Palate; Patients; Phenotype; Process; Proteomics; Scaffolding Protein; Smoking; Source; Specific qualifier value; Stains; Syndrome; System; Therapeutic; Three-dimensional analysis; Time; Tissues; cell growth regulation; cleft lip and palate; dietary; dosage; experimental study; folic acid supplementation; improved; in utero; innovation; insight; intraperitoneal; mouse model; mutant; non-muscle myosin; novel; orofacial cleft; palatal shelves; palatogenesis; phosphoproteomics; pregnant; prevent; translational potential

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. Isolated cleft palate (CP) alone accounts for 1/1700 cases in the US. In addition, there are >400 syndromes with CP as a phenotype. Environmental factors such as folate intake and smoking have been shown to affect maternal environment. While dietary maternal folic acid supplementation has consistently shown ameliorative effects on neural tube defects, its effect on reducing the occurrence of CL/P has been variable both in human and in mouse models. Sources of this variability include the dosage and delivery of folic acid as well as ability to assess the impact of the treatment in mouse models. Several human studies have speculated that low folic acid dose is not sufficient to prevent CL/P, and that higher doses are more effective. Our preliminary data indicate that a high daily dose of folic acid given intraperitoneally to pregnant mice in two short developmental windows is sufficient to rescue CP in utero in Specc1l mouse model of CP. Specc1l deficiency leads to actin cytoskeletal defects, which results in palatal shelf elevation delay. We have developed several quantitative 2-D and 3-D analyses to characterize the timing and cellular changes that occur during palatal shelf growth and elevation. We posit that in utero folic acid supplementation has an ameliorative effect on the dynamics of palate closure. In contrast to neural tube defects, we argue that rescue of CP requires a higher daily dose of folic acid administered intraperitoneally but for a short window of time. To our knowledge, we are the first to use intraperitoneal injection of folic acid to rescue CP. Folic acid has been used extensively in human studies and has been safe even at relatively high doses. The objective of this proposal is to determine the minimum possible dose and treatment window for an effective rescue of CP and to study which aspects of palate closure dynamics are impacted by folic acid supplementation. We will also use sophisticated proteomic and phosphoproteomic analysis of palatal shelf tissue to determine the molecular consequence of folic acid treatment. Together, these studies will establish the experimental system and methods to study maternal folic acid rescue of CP in mice that can be applied to other mouse models. Importantly, these studies will lead to future therapeutic strategies to reduce the incidence of CL/P in humans.

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。在美国，仅孤立性腭裂(CP)就占1700例中的1例。在……里面 此外，还有以CP为表型的&gt；400综合征。环境因素，如叶酸摄入量和 吸烟已被证明会影响母体环境。而孕妇膳食中的叶酸补充 一直显示出对神经管缺陷的改善作用，在减少CL/P的发生方面的作用 在人类和小鼠模型中都是可变的。这种变异性的来源包括剂量和投放 以及评估治疗对小鼠模型的影响的能力。几项人体研究 曾推测，低剂量的叶酸不足以预防CL/P，而高剂量的叶酸 有效。我们的初步数据表明，怀孕小鼠每天给予大剂量叶酸 在两个较短的发育窗内，就足以挽救Cp小鼠模型子宫内的Cp。规格1L 缺失导致肌动蛋白细胞骨架缺陷，从而导致腭架抬高延迟。我们已经开发出 几个定量的2-D和3-D分析，以表征时间和细胞变化 腭架的生长和抬高。我们假设在子宫内补充叶酸有改善作用。 关于上颌闭合的动力学。与神经管缺陷不同，我们认为抢救CP需要一个 每天给予较高剂量的叶酸，但持续时间较短。据我们所知， 我们首次使用叶酸腹腔注射抢救慢性胰腺炎。叶酸已被广泛用于 人体研究，即使在相对较高的剂量下也是安全的。这项提案的目标是确定 有效抢救CP的最小可能剂量和治疗窗口，并研究 补充叶酸会影响上颌闭合的动力。我们还将使用复杂的蛋白质组学 以及对腭架组织进行磷酸蛋白质组学分析以确定叶酸的分子后果 治疗。总之，这些研究将建立研究母体叶酸的实验系统和方法 酸挽救小鼠的CP，可应用于其他小鼠模型。重要的是，这些研究将导致 未来减少人类CL/P发生率的治疗策略。