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.

项目摘要/摘要 怀孕期间的饮酒可能导致胎儿酒精疾病（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有关，并且在没有的情况下很难诊断 面部特征。产前酒精暴露（PAE）会对发育中的胎儿产生全身影响，但 大多数研究都是针对大脑神经元特异性变化及其对PAE-的贡献进行的研究。 相关的神经发育障碍。很少有PAE研究探索了酒精引诱的改变 发育过程中大脑的脉管系统。 microRNA介导的机制，导致变化 PAE期间的脑脉管系统尚不清楚。我们的初步研究表明miR-150-5p上调 在PAE期间，在脑微血管内皮细胞（BMVEC）中。这导致其监管目标减少， 包括血管内皮锌指1（VEZF1），这是我们确定的新目标，这是一个关键的调节剂 血管发育。我们还测试了miR-150-5p对内皮细胞功能的影响。我们有 使用的迁移，管形成和渗透性测定是一种分析血管生成和血脑的一种方式 体外障碍完整性。初步数据表明，过表达miR-150-5p的迁移和 管形成，并增加了BMVEC渗透性，而miR-150-5p抑制作用也相反。此外， 用酒精治疗细胞导致miR-150-5p表达增加，这也导致下降 迁移和管形成。我们的初步研究强调了miR-150-5p对调节的重要性 内皮细胞行为和酒精对内皮细胞功能的不利影响。自从micrornas以后 转录调节许多不同的mRNA靶标，我们假设miR-150-5p抑制了血管 对PAE期间皮质中皮质的血管形态和功能的不利影响。以下目标将 进行研究以检验以下假设：1）确定由miR-150-5p抑制的靶标在原代BMVEC中 在产前酒精暴露期间，2）确定miR-150-5p抑制和/或目标过表达的影响 在产前酒精暴露的胚胎的一级BMVEC上，3）检查miR-150-5p的体内效应 抑制和/或目标干扰PAE期间发育中的皮质脉管系统。成功完成 该项目集成的目标将更好地了解病理中的血管贡献 Pae。此外，Ruth L. Kirschstein国家研究服务奖的支持 促进与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 致力于独立研究。