Maternal-fetal amino acid transfer across placenta in a mouse model of prenatal alcohol exposure

项目总结/摘要 此补充的目的是申请资助，4个月的延长我目前的F32奖，从 二零二一年八月一日至二零二一年十二月三十一日。由于COVID-19大流行，我无法继续 进行我的F32项目目标2中描述的实验，因为我目前的F32项目 并没有被我的机构指定为必要研究。此外，我是一个单身母亲， 大流行导致我孩子的学校和当地的许多课外活动关闭， 社区没有托儿服务，我不得不花大量的时间在家教育我的孩子， 这增加了我照顾孩子的责任，也带来了巨大的困难，严重阻碍了我的研究。 中求进工作总申请的延期将补偿因大流行而损失的研究时间， 二零二零年三月至二零二零年七月。在这4个月的延长期内，我将完成免疫组化实验 其研究了暴露于酒精的胎盘中氨基酸（AA）转运蛋白的细胞分布， 如目标2所述，这些实验将完成一份手稿。我也会分析完 从这些暴露于酒精的胎盘中生成的转录组数据集，另一个实验在Aim中描述 2、第二个手稿是接近完成。申请的资金将支付我在这4- 一个月的延长期，完成免疫组化所需的研究用品，以及费用 发表报告这些结果的手稿。总体项目假设保持不变， 指出产前酒精暴露（PAE）导致宫内生长受限，至少部分是通过减少 胎盘AA供应给胎儿，这是下调胎盘mTOR信号传导的结果， AA运输和AA代谢改变。这三个具体目标也保持不变。目标1执行 全面的代谢组学分析，以表征PAE如何降低AA供应和代谢结局 沿着母体-胎盘-胎儿轴。目的2进行转录组学分析，蛋白质印迹， 胎盘中的免疫组织化学来检验胎盘AA转运蛋白的下调 代谢基因导致PAE中AA水平的改变。Aim 3在 胎盘，以检验AA转运和代谢的这些变化伴随以下假设： 抑制胎盘mTOR通路，其是AA可用性和胎儿生长的主要调节剂。这些 研究使用尖端技术来创建PAE如何影响胎盘AA供应的全球画像， 为PAE如何导致宫内生长受限提供了新的机制见解。请求 延期将使我能够完成拟议的F32研究，为未来的研究奠定基础 检查出生后的神经和代谢健康，遗传风险因素的调节作用， 母亲补充AA对改善PAE妊娠结局的有效性。

Project Summary/Abstract The purpose for this supplement is to request a funded, 4-month extension for my current F32 award, from August 1st, 2021 to December 31st, 2021. Due to the COVID-19 pandemic, I was unable to continue performing experiments described in Aim 2 of my F32 project because the project under my current F32 award was not designated by my institution as essential research. Furthermore, I am a single mother with a young child, and the pandemic led to the shutdown of my child’s school and many afterschool programs in the local community. Without access to childcare, I had to spend a large amount of time homeschooling my child, which created an additional childcare responsibility and tremendous hardships that significantly hindered my research progress. The requested extension will compensate for the research time loss due to the pandemic, from March 2020 to July 2020. During this 4-month extension, I will complete the immunohistochemical experiments which investigate the cellular distribution of the amino acid (AA) transporters in the alcohol-exposed placentas, as described in Aim 2; these experiments would wrap up a manuscript. I will also finish analyzing the transcriptomic dataset generated from these alcohol-exposed placentas, another experiment described in Aim 2, for a second manuscript that is near-completion. The requested funds will cover my stipends during this 4- month extension period, research supplies needed to complete the immunohistochemistry, and fees for publishing the manuscripts reporting these results. The overall project hypothesis remains unchanged, which states that prenatal alcohol exposure (PAE) causes intrauterine growth restriction, at least in part, by reducing placental AA supply to the fetus, and that this is a consequence of downregulated placental mTOR signaling, AA transport, and altered AA metabolism. The three specific aims also remain unchanged. Aim 1 performs a comprehensive metabolomics analysis to characterize how PAE decreases AA supply and metabolic fate along the maternal-placental-fetal axis. Aim 2 performs transcriptomics analysis, western blotting and immunohistochemistry in the placenta to test the hypothesis that downregulation of placental AA transporters and metabolic genes contributes to the altered AA levels in PAE. Aim 3 performs western blotting in the placenta to test the hypothesis that these changes in AA transport and metabolism are accompanied by inhibition of placental mTOR pathways, which is a major regulator of AA availability and fetal growth. These studies use cutting-edge techniques to create a global portrait of how PAE affects placental AA supply and offer novel mechanistic insight into how PAE contributes to intrauterine growth restriction. The request extension will allow me to finish the proposed F32 research studies, which lay groundwork for future research that examines postnatal neural and metabolic health, the modulatory effect of genetic risk factors, and the effectiveness of maternal AA supplementation to improve outcomes of PAE pregnancies.