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。在美国，仅孤立性腭裂 (CP) 病例就占 1/1700。在 此外，还有超过 400 种以 CP 为表型的综合征。环境因素，例如叶酸摄入量和 吸烟已被证明会影响母亲的环境。母亲在膳食中补充叶酸的同时 一直显示出对神经管缺陷的改善作用，其对减少 CL/P 发生的作用 在人类和小鼠模型中都存在差异。这种变异性的来源包括剂量和递送 叶酸的含量以及评估治疗对小鼠模型的影响的能力。多项人体研究 推测低叶酸剂量不足以预防 CL/P，而较高剂量则更有效。 有效的。我们的初步数据表明，每天给怀孕小鼠腹腔注射高剂量的叶酸 在 CP 的 Specc1l 小鼠模型中，在两个较短的发育窗口内足以挽救子宫内的 CP。规格1l 缺乏会导致肌动蛋白细胞骨架缺陷，从而导致腭架抬高延迟。我们开发了 一些定量的 2-D 和 3-D 分析来表征过程中发生的时间和细胞变化 腭架生长和抬高。我们认为在子宫内补充叶酸具有改善作用 上颚闭合的动力学。与神经管缺陷相反，我们认为挽救 CP 需要 腹腔内给予叶酸的每日剂量较高，但时间窗口较短。据我们所知， 我们率先采用腹腔注射叶酸来抢救CP。叶酸已广泛应用于 人类研究表明，即使在相对较高的剂量下也是安全的。该提案的目的是确定 有效挽救脑瘫的最小可能剂量和治疗窗口，并研究哪些方面 上颚闭合动力学受到叶酸补充的影响。我们还将使用复杂的蛋白质组学 对腭架组织进行磷酸蛋白质组学分析，以确定叶酸的分子结果 治疗。这些研究将共同​​建立研究母体叶酸的实验系统和方法 对小鼠 CP 的酸拯救可应用于其他小鼠模型。重要的是，这些研究将导致 降低人类 CL/P 发病率的未来治疗策略。