Identification, Quantification, and Functional Characterization of Transporters in Human Placenta, Developing Gut and Fetal Brain

人胎盘、肠道和胎儿大脑发育中转运蛋白的鉴定、定量和功能表征

• 批准号: 10746192
• 负责人: JASHVANT D Unadkat
• 金额: $ 92.16万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-11 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10746192
• 关键词: ABCB1 gene; ABCG2 gene; ATP-Binding Cassette Transporters; Address; Adult; Articulation; Binding; Biopsy; Birth; Blood - brain barrier anatomy; Brain; Breast Feeding; Carrier Proteins; Cells; Childhood; Clinical; Data; Development; Drug or chemical Tissue Distribution; Exposure to; FDA approved; Family; Fetal Development; Fetus; First Pregnancy Trimester; Folic Acid; Gastrointestinal tract structure; Goals; Growth; HIV; HIV Protease Inhibitors; Human; Human Development; Human Genome; Immunohistochemistry; In Vitro; Infant; Ingestion; Intestines; Knowledge; Mediating; Messenger RNA; Modeling; Mothers; Nutrient; Organ; Organoids; Perfusion; Pharmaceutical Preparations; Physiological; Placenta; Play; Pluripotent Stem Cells; Pregnancy; Pregnant Women; Proteins; Proteomics; Role; SLC19A1 gene; Saquinavir; Second Pregnancy Trimester; Tissues; Toxic effect; Transfection; Treatment Efficacy; Universities; Vascularization; Vitamins; Vulnerable Populations; Washington; Xenobiotics; absorption; brain endothelial cell; dietary supplements; drug efficacy; fetal; fetal blood; human tissue; induced pluripotent stem cell; interest; mRNA Expression; microphysiology system; neonate; novel; prenatal exposure; solute; uptake

SUMMARY The placenta, often referred to as the “intestine” of the fetus, is an essential organ that controls the exchange of nutrients (including vitamins) and xenobiotics (i.e., dietary supplements and FDA approved drugs) between the mother and her fetus. The fetus can also ingest nutrients and xenobiotics via the digestive tract, so do neonates and infants through breastfeeding after birth. The fetal, neonate, and infant blood-brain barrier (BBB) serves a critical role in protecting the developing brain from xenobiotics and supplying nutrients to the brain. Thus, the placenta, the developing BBB and gut are key organs responsible for nutrient and xenobiotic distribution and absorption impacting early human development and xenobiotic toxicity. Transporters can play an essential role in the absorption, systemic exposure, and tissue distribution of nutrients and xenobiotics in the fetus, neonates, and infants across the placental, intestinal, and blood-brain barriers. Identification and quantification of transporters in these tissue barriers is important for understanding and predicting fetal or neonate/infant uptake of, and exposure to, nutrients and xenobiotics, and hence impacting early development as well as the safe and efficacious use of medications/supplements in these vulnerable populations. While the expression and function of a few ABC transporters in human term placenta, such as P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP), have been well-delineated, such data are sorely missing for transporters in early gestation placenta, and in the developing gut and BBB during pregnancy and after birth. In this application, we propose to establish a Transporter Elucidation Center (TEC) at the University of Washington that addresses the goals articulated in RFA-HD-23-003. Using quantitative global and targeted proteomics, we will systematically identify and quantify the ontogeny of transporters in the human placenta (from early gestation to term), the developing gut as well as the developing BBB (from early and mid-gestation and after birth). Then, through in vitro (transporter-transfected cells, immunohistochemistry, immunolocalization) and ex vivo (e.g., placental perfusion, intestinal organoids, and iPSC-derived human fetal BBB models) transport studies, we will determine novel substrates, cellular localization, and transport activity of highly abundant transporters in these tissues. Combined, these studies will address a critical knowledge gap in our understanding of transporters that control essential physiological functions and xenobiotic disposition in the developing fetus and neonate/infant. Consequently, the proposed studies will enhance our ability to predict the toxicity or efficacy of xenobiotics and physiological efficacy of nutrients (or lack thereof) in these vulnerable populations.

总结 胎盘，通常被称为胎儿的“肠”，是控制胎儿与胎儿之间的交换的重要器官。 营养物（包括维生素）和异生物质（即，膳食补充剂和FDA批准的药物）之间 母亲和胎儿胎儿也可以通过消化道摄取营养物质和外源性物质，新生儿也是如此 和婴儿出生后通过母乳喂养。胎儿、新生儿和婴儿血脑屏障（BBB）是一个重要的屏障。 在保护发育中的大脑免受外源性物质的影响和向大脑提供营养方面起着关键作用。因此 胎盘、发育中的血脑屏障和肠道是负责营养和异生物质分布的关键器官， 影响人类早期发育和异生物质毒性的吸收。运输者可以发挥重要作用， 在营养素和外源性物质的吸收、全身暴露和组织分布中的作用 胎儿、新生儿和婴儿通过胎盘、肠和血脑屏障。识别和 定量这些组织屏障中的转运蛋白对于理解和预测胎儿或新生儿的发育是重要的。 新生儿/婴儿摄取和暴露于营养素和外源性物质，因此影响早期发育， 以及在这些弱势群体中安全有效地使用药物/补充剂。而 人胎盘组织中一些ABC转运蛋白的表达和功能，如P-糖蛋白（P-gp）和 乳腺癌耐药蛋白（BCRP），已经被很好地描述，这样的数据是非常缺乏的， 在妊娠早期胎盘中，以及在妊娠期间和出生后的发育中的肠道和BBB中的转运蛋白。在 本申请中，我们建议在华盛顿大学建立一个转运蛋白阐明中心（TEC 该文件旨在实现RFA-HD-23-003中阐述的目标。使用定量的全局和靶向蛋白质组学， 我们将系统地识别和量化人类胎盘中转运蛋白的个体发育（从早期到晚期）， 妊娠至足月）、发育中的肠道以及发育中的BBB（从妊娠早期和中期以及之后 出生）。然后，通过体外（转运蛋白转染细胞、免疫组织化学、免疫定位）和体外（体外）实验， 体内（例如，胎盘灌注、肠类器官和iPSC衍生的人胎儿BBB模型）转运 研究，我们将确定新的底物，细胞定位，和运输活动的高度丰富的 这些组织中的转运蛋白。结合起来，这些研究将解决我们的关键知识差距， 了解控制基本生理功能和外源性处置的转运蛋白， 发育中的胎儿和新生儿/婴儿。因此，拟议的研究将提高我们预测 外源性物质的毒性或功效以及营养素的生理功效（或缺乏营养素）在这些脆弱的 人口。