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

项目总结/摘要 怀孕期间饮酒可能导致胎儿酒精谱系障碍（FASD）， 综合征（FAS）是最严重的形式，表现为面部异常。较轻形式的 FASD与广泛的神经认知缺陷相关，并且在缺乏神经认知缺陷的情况下难以诊断。 面部特征产前酒精暴露（PAE）对发育中的胎儿产生全身性影响， 大多数研究都是针对大脑中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研究探讨了酒精引起的改变， 大脑的血管系统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- 5 p上调， 脑微血管内皮细胞（BMVECs）在PAE。这导致其调节目标减少， 包括血管内皮锌指1（Vezf 1），我们已经确定的一个新的目标，这是一个关键的调节因子， 血管的发育。我们还测试了miR-150- 5 p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- 5 p减少了迁移， 管形成，并且它增加BMVEC渗透性，而miR-150- 5 p抑制则相反。此外，本发明还 用酒精处理细胞导致miR-150- 5 p表达增加，这也导致miR-150- 5 p表达减少。 迁移和管形成。我们的初步研究强调了miR-150- 5 p在调节细胞凋亡中的重要性。 内皮细胞行为和酒精对内皮细胞功能的不良影响。由于microRNA后- 由于miR-150- 5 p转录调节许多不同的mRNA靶点，我们假设miR-150- 5 p抑制血管生成， 靶向对PAE期间皮质中的血管形态和功能产生不利影响。以下目标将 1）鉴定在原代BMVEC中被miR-150- 5 p抑制的靶标 在产前酒精暴露期间，2）确定miR-150- 5 p抑制和/或靶向过表达的作用 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- 5 p的体内作用 抑制和/或靶向干扰在PAE期间发育的皮质脉管系统。成功完成 本项目的目的将使我们更好地理解血管在病理学中的作用。 的PAE。此外，Ruth L.基尔施斯坦国家研究服务奖个人博士前 促进健康相关研究多样性的奖学金将为我的职业轨迹做出重大贡献 走向独立研究。

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.