miR-150-5p Disrupts Vascular Development in the Brain during Prenatal Alcohol Exposure

miR-150-5p 在产前酒精暴露期间破坏大脑血管发育

• 批准号: 10312695
• 负责人: Gabriela Perales
• 金额: $ 3.06万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10312695
• 关键词: 3' Untranslated Regions; Adverse effects; Affect; Alcohols; Automobile Driving; Binding; Binding Sites; Biological Assay; Blood - brain barrier anatomy; Blood Vessels; Brain; Cell physiology; Cells; Cerebrovascular system; Characteristics; Child; Data; Defect; Development; Diagnosis; Electrical Resistance; Embryo; Endothelial Cells; Ethanol; Face; Fellowship; Fetal Alcohol Exposure; Fetal Alcohol Spectrum Disorder; Fetal Alcohol Syndrome; Fetal Development; Fetal alcohol effects; Genes; Genetic Transcription; Health; In Vitro; Individual National Research Service Award; Injections; Lead; Literature; Luciferases; Mediating; Messenger RNA; MicroRNAs; Microscopy; Molecular; Morphology; Mothers; National Research Service Awards; Neurocognitive Deficit; Neurodevelopmental Disorder; Neuronal Dysfunction; Neurons; Pathology; Pathway interactions; Patients; Pattern; Permeability; RNA; Regulation; Reporting; Research; School-Age Population; Techniques; Testing; Therapeutic Intervention; Transcript; Tube; Vascular Endothelium; Work; Zinc Fingers; alcohol consumption during pregnancy; alcohol effect; angiogenesis; base; blastomere structure; brain endothelial cell; career; cell behavior; differential expression; embryonic alcohol exposure; in utero; in vivo; knock-down; migration; mouse model; neuron development; novel; overexpression; pre-doctoral; prenatal; pup; transcriptome sequencing; vascular abnormality; vascular contributions

PROJECT SUMMARY/ABSTRACT Alcohol consumption during pregnancy may result in fetal alcohol spectrum disorders (FASD), with fetal alcohol syndrome (FAS) being the most severe form and manifesting with facial abnormalities. Less severe forms of FASD are associated with a wide range of neurocognitive deficits and are difficult to diagnose in the absence of facial characteristics. Prenatal alcohol exposure (PAE) produces systemic impacts on the developing fetus, but most studies have been conducted on neuronal-specific changes in the brain and their contribution to PAE- associated neurodevelopmental disorders. Very few PAE studies have explored alcohol-elicited alterations to the vasculature of the brain during development. microRNA-mediated mechanisms that result in alterations to the brain vasculature during PAE are unknown. Our preliminary studies indicate that miR-150-5p is upregulated in brain microvascular endothelial cells (BMVECs) during PAE. This results in a decrease in its regulatory targets, including vascular endothelial zinc finger 1 (Vezf1), a novel target we have identified, which is a critical regulator of vascular development. We have also tested the effects of miR-150-5p on endothelial cell function. We have used migration, tube formation, and permeability assays as a way to analyze angiogenesis and blood brain barrier integrity in vitro. Preliminary data indicated that overexpressing miR-150-5p decreased migration and tube formation, and it increased BMVEC permeability, while miR-150-5p inhibition did the opposite. Additionally, treating cells with alcohol resulted in an increase in miR-150-5p expression which also resulted in decreased migration and tube formation. Our preliminary studies underscore the importance of miR-150-5p on regulating endothelial cell behavior and the adverse effects alcohol has on endothelial cell function. Since microRNAs post- transcriptionally regulate many different mRNA targets, we hypothesized that miR-150-5p is inhibiting vascular targets to adversely affect vascular morphology and function in the cortex during PAE. The following aims will be investigated to test this hypothesis: 1) Identify targets that are inhibited by miR-150-5p in primary BMVECs during prenatal alcohol exposure, 2) Determine the effects of miR-150-5p inhibition and/or target overexpression on primary BMVECs from prenatal alcohol exposed embryos, and 3) Examine the in vivo effects of miR-150-5p inhibition and/or target interference on the developing cortical vasculature during PAE. Successful completion of the aims integrated in this project will provide a better understanding of vascular contributions in the pathology of PAE. In addition, support from the Ruth L. Kirschstein National Research Service Award Individual Predoctoral Fellowship to Promote Diversity in Health-Related Research will contribute significantly to my career trajectory towards independent research.

项目概要/摘要 怀孕期间饮酒可能导致胎儿酒精谱系障碍（FASD），胎儿酒精 综合征（FAS）是最严重的形式，表现为面部异常。不太严重的形式 FASD 与多种神经认知缺陷相关，在缺乏相关信息的情况下很难诊断 面部特征。产前酒精暴露（PAE）会对发育中的胎儿产生系统性影响，但是 大多数研究都是针对大脑神经元特异性变化及其对 PAE 的贡献进行的。 相关的神经发育障碍。很少有 PAE 研究探索酒精引起的改变 发育过程中大脑的脉管系统。 microRNA介导的机制导致改变 PAE 期间的脑血管系统尚不清楚。我们的初步研究表明 miR-150-5p 上调 PAE 期间脑微血管内皮细胞 (BMVEC) 中的变化。这导致其监管目标下降， 包括血管内皮锌指 1 (Vezf1)，这是我们确定的一个新靶标，它是一个关键的调节因子 血管的发育。我们还测试了 miR-150-5p 对内皮细胞功能的影响。我们有 使用迁移、管形成和渗透性测定作为分析血管生成和血脑的方法 体外屏障完整性。初步数据表明，过表达 miR-150-5p 会减少迁移和 管形成，并增加 BMVEC 通透性，而 miR-150-5p 抑制则相反。此外， 用酒精处理细胞会导致 miR-150-5p 表达增加，从而导致 miR-150-5p 表达减少 迁移和管形成。我们的初步研究强调了 miR-150-5p 在调节方面的重要性 内皮细胞行为以及酒精对内皮细胞功能的不利影响。由于 microRNA 后 转录调节许多不同的 mRNA 靶标，我们假设 miR-150-5p 抑制血管 目标是在 PAE 过程中对皮质血管形态和功能产生不利影响。将实现以下目标 进行研究以检验该假设：1) 鉴定原代 BMVEC 中 miR-150-5p 抑制的靶标 产前酒精暴露期间，2) 确定 miR-150-5p 抑制和/或目标过度表达的影响 对来自产前酒精暴露胚胎的原代 BMVEC 的影响，以及 3) 检查 miR-150-5p 的体内作用 PAE 期间对发育中的皮质脉管系统的抑制和/或目标干扰。顺利完成 该项目的目标将有助于更好地了解血管在病理学中的作用 PAE 的。此外，还得到了 Ruth L. Kirschstein 国家研究服务奖个人博士前的支持 促进健康相关研究多样性的奖学金将对我的职业轨迹做出重大贡献 走向独立研究。