Uncovering the cellular and molecular mechanisms of folic acid fortification in neural tube defects

揭示叶酸强化治疗神经管缺陷的细胞和分子机制

基本信息

批准号：
10595548
负责人：
Anneke D Kakebeen
金额：
$ 7.2万
依托单位：
UNIVERSITY OF COLORADO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10595548
关键词：
Address Affect Articulation Back Bioinformatics Brain Candidate Disease Gene Cell physiology Cells Chick Chick Embryo Chromatin Clinical Collaborations Communication Communities Compensation Complex Congenital Abnormality Congenital Disorders Data Analyses Data Set Deposition Development Diet Dimensions Disease Educational process of instructing Embryo Embryology Environmental Risk Factor Etiology Extracellular Matrix Failure Family Fetal Mortality Statistics Folic Acid Gene Expression Gene Expression Profile Gene Mutation Genes Genetic Genetic Transcription Histological Techniques Histology Human Image Incidence Knock-out Lead Learning Mentors Mentorship Modeling Molecular Morphogenesis Movement Multiomic Data Mus Mutant Strains Mice Neural Tube Closure Neural Tube Defects Neural tube Pattern Persons Play Population Pregnancy Preparation Prevention Prevention strategy Process Public Health Research Research Personnel Risk Risk Assessment Signal Transduction Spinal Cord Students System Techniques Testing Training United States Validation Work candidate validation cell behavior cell fixing cell motility cell type cellular imaging de novo mutation design environmental stressor experimental study folic acid supplementation fortification gene discovery gene environment interaction gene regulatory network genetic testing high risk improved information model innovation insight meetings mouse genetics mutant mutant mouse model null mutation pregnant prenatal prevent provider communication risk variant screening single-cell RNA sequencing skills teacher transcription factor undergraduate research undergraduate research experience undergraduate student

项目摘要

SUMMARY The objective of this proposal is to elucidate how folic acid prevents neural tube defects on the cellular and molecular level, and to develop a platform to functionally validate and test the FA responsiveness of human NTD candidate genes. Neural tube defects (NTDs) are the second most common congenital abnormality, affecting 6 in 10,000 pregnancies in the United States. Failure to close the neural tube, the precursor to the brain and spinal cord, is complex disorder resulting from genetic and environmental stressors. Because multiple factors play into the development of this disorder, it is critical to understand how environmental factors interact with genetics in the etiology of NTDs. While population-wide folic acid (FA) fortification has reduced the overall NTD rate by 28% in the US, it remains unclear how folic acid prevents NTDs. My proposal aims to address two hypotheses about folic acid and neural tube defects. First, I hypothesize that FA contributes to NTD prevention by acting at the cell movement level as well as inducing transcriptional cascades to compensate for the dysregulated genes. To uncover whether FA can rescue NT closure by acting at the cellular or molecular level, I will characterize the cellular movements and transcriptional profiles for the Alx1 mutant mouse model supplemented with FA. The Alx1 knockout model results in NTDs in 100% of mutant embryos, however prenatal FA supplementation can reduce the incidence to 13%. The Alx1 mutant is a tractable model to dissect the contribution of FA fortification to restoring NT closure because NTDs in this model result from dysregulation of a transcription factor and abnormal cellular movements. We will use imaging-based techniques to characterize cell morphogenesis processes and scRNA-Seq and scATAC-Seq to identify FA-induced gene expression in the Alx1-null background. Results from these complimentary studies will inform if FA acts by supporting cellular movements, inducing transcriptional activity, or a combination. Second, I hypothesize that testing more gene mutations for FA responsiveness will reveal patterns of gene types that benefit from FA fortification as a prevention strategy. To address this hypothesis, I will develop a course-based undergraduate research experience (CURE) entitled The What, the Why, and the How of neural tube defects. The scientific aim of this course will be to functionally validate candidate human NTD genes of unknown significance in NT closure in chick embryos and test the folic acid responsiveness. The societal aim of the CURE is to increase the number of relevant research opportunities for undergraduates while meeting a public health need to screen candidate genes and communicate back to clinicians and families. I expect screening more candidate genes for NTDs and FA responsiveness will reveal new high-risk genes and increase our predictive power over the types of dysregulated genes likely responsive to FA supplementation. Together, the results from this proposal will be important to bolster our understanding of how FA can prevent NTDs and improve risk assessment of preventative strategies against neural tube defects.

概括该建议的目的是阐明叶酸如何防止细胞上的神经管缺陷分子水平，并开发一个平台以在功能上验证和测试人类NTD的FA响应能力候选基因。神经管缺陷（NTD）是第二个常见的先天性异常，影响6 在美国的10,000例怀孕中。无法关闭神经管，这是大脑和脊柱的前体绳索是由遗传和环境压力源引起的复杂疾病。因为多种因素涉及到这种疾病的发展，了解环境因素如何与遗传学相互作用至关重要 NTD的病因。虽然整个人口叶酸（FA）的强化使总NTD率降低了28％在美国，尚不清楚叶酸如何阻止NTD。我的提议旨在解决两个关于叶酸和神经管缺陷。首先，我假设FA通过在细胞上作用有助于NTD预防运动水平以及诱导转录级联反应以补偿失调的基因。到发现FA是否可以通过在细胞或分子水平上作用来挽救NT闭合，我将表征补充FA的ALX1突变小鼠模型的细胞运动和转录曲线。这 ALX1敲除模型导致100％突变胚胎的NTD，但是补充产前FA可以将发病率降低到13％。 ALX1突变体是一种可剖析FA强化贡献的可寓言模型恢复NT闭合，因为该模型中的NTD是由于转录因子的失调和细胞运动异常。我们将使用基于成像的技术来表征细胞形态发生过程以及SCRNA-SEQ和SCATAC-SEQ，以鉴定Alx1-Null中FA诱导的基因表达背景。这些免费研究的结果将告知FA是否通过支持细胞运动来行动，诱导转录活性或组合。其次，我假设测试更多的基因突变 FA响应能力将揭示受益于FA强化作为预防策略的基因类型的模式。为了解决这一假设，我将开发基于课程的本科研究经验（CURE）神经管缺陷的原因，原因和方式。本课程的科学目的是在功能上验证候选人NTD基因在雏鸡胚胎中NT闭合意义不明的基因并测试叶面酸反应能力。治愈的社会目的是增加相关研究机会的数量对于本科生满足公共卫生需要筛查候选基因并与临床医生和家庭。我希望筛选更多的NTD候选基因，而FA响应能力将揭示新的高风险基因，并提高我们对失调基因类型的预测能力可能响应的补充FA。总之，该提案的结果对于增强我们对的理解将很重要 FA如何防止NTD并改善针对神经管缺陷的预防策略的风险评估。