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。在美国，单独的腭裂（CP）占1/1700例。在 此外，有>400种综合征以CP作为表型。环境因素，如叶酸摄入量和 吸烟已被证明会影响母体环境。虽然母亲的膳食补充叶酸 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 以及评估小鼠模型中治疗影响的能力。几项人体研究 据推测，低剂量叶酸不足以预防CL/P，高剂量叶酸更能预防CL/P。 有效我们的初步数据表明，每天高剂量的叶酸给孕鼠腹腔注射， 在两个较短的发育窗口中足以在CP的Specc 11小鼠模型中拯救子宫内的CP。Specc1l 缺乏导致肌动蛋白细胞骨架缺陷，这导致腭架升高延迟。我们已经开发 几个定量的2-D和3-D分析，以表征时间和细胞的变化，发生在 腭架生长和升高。我们认为，在子宫内补充叶酸具有改善作用， 上腭闭合的动力学与神经管缺陷相反，我们认为CP的拯救需要一个 腹膜内给予较高的叶酸日剂量，但时间窗较短。据我们所知， 我们是第一个使用腹腔注射叶酸来抢救CP的。叶酸已广泛用于 人类研究，即使在相对高的剂量下也是安全的。本提案的目的是确定 有效挽救CP的最小可能剂量和治疗窗口，并研究 腭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 和腭架组织的磷酸蛋白质组学分析，以确定叶酸的分子后果 治疗这些研究将为建立研究母体叶酸的实验体系和方法奠定基础 小鼠中CP的酸拯救，其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。