Transporter Elucidation Center at the University of California, San Francisco

加州大学旧金山分校转运蛋白阐明中心

基本信息

批准号：
10747230
负责人：
KATHLEEN M GIACOMINI
金额：
$ 96.9万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-19 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10747230
关键词：
ATP-Binding Cassette Transporters Address Antibodies Binding Biological Assay Biological Process Biology Blood - brain barrier anatomy Blood capillaries Brain California Capillary Endothelial Cell Categories Cell Line Cell model Cerebrovascular system Chemicals Communities Computational Biology Cryoelectron Microscopy Data Docking Drug Prescriptions Drug Targeting Endothelial Cells Epithelial Cells Epitopes Fetal Development G-Protein-Coupled Receptors Gatekeeping Gene Family Goals Homeostasis Hormones Human Human Genome Immunohistochemistry Infant Informatics Knowledge Libraries Ligands Macronutrients Nutrition Mammary gland Membrane Membrane Transport Proteins Mitochondria Modeling Molecular Structure Movement Nutrient Opioid Orphan Pharmaceutical Preparations Pharmacology Placenta Play Poison Proteomics Psychotropic Drugs Recombinants Research Research Personnel Research Support Resources Role Sampling San Francisco Scientist Sea Signaling Molecule Specificity Structural Models Structure System Testing Tight Junctions Tissues Transmembrane Transport Transport Process Universities Work Xenobiotics blood-brain barrier function cerebral capillary comparative dietary constituent dietary supplements drug discovery in silico inhibitor interest member multidisciplinary polarized cell programs protein data bank protein structure psychostimulant response solute structural biology tool virtual screening

项目摘要

PROJECT SUMMARY Membrane transporters in the Solute Carrier (SLC) and ATP Binding Cassette (ABC) superfamilies play essential roles in the transmembrane movement of solutes including drugs, dietary supplements and nutrients. Transporters on the polarized membranes of epithelial cells such as those in the placenta, mammary gland and gut or polarized endothelial cells such as those of the Blood Brain Barrier (BBB) play essential roles in transcellular flux of their substrates. A significant gap in our understanding of both SLC and ABC transporters exists as many transporters have not been systematically characterized and the range of substrates is not known or poorly understood. In response to RFA-HD-23-003, we propose to establish a multi-disciplinary Transporter Elucidation Center at the University of California San Francisco (TECUCSF), which brings together expert scientists in transporter biology and pharmacology, membrane transporter structural biology, computational biology, and chemo-informatics. The overall goal of the TECUCSF is to elucidate the ligand specificity, protein structure and cellular localization of SLC and ABC transporters in the human BBB. Based on preliminary studies from our global proteomic analyses, we will prioritize a list of 30 highly expressed and understudied/orphan SLC and ABC transporters in the human BBB (BBB-30). Key knowledge gaps of these 30 transporters will be addressed by research conducted under three aims. For Aim 1, we will perform in silico docking of neuroactive drugs, nutrients, and dietary supplements using experimentally determined structures or structures based on computed structural models of transporters in the BBB-30. For each of these transporters, we will create recombinant cell lines expressing the transporter and functional assays to validate the in silico results, which will result in the discovery of ligands for both understudied and orphan transporters. For Aim 2, we will use cryoEM to determine the structure of selected transporters in the BBB-30, which do not have available structures. Finally, for Aim 3, we will perform localization of the BBB-30 in human endothelial cell models using available antibodies and flux studies will be performed to confirm the localization. To support our studies and enhance studies in the Transporter Elucidation Network (TEN), three cores will be established: TECUCSF Informatics and Modeling Core; TECUCSF Structure Core; and TECUCSF Function Core. The cores will provide support for research proposed in each of the three aims, as well for investigators in the TEN. TECUCSF will work together with other TECs within the TEN to generate knowledge and resources that will be shared with the broader research community through RESOLUTE to advance our understanding of nutrient, drug, and dietary supplement constituent transport across the BBB.

项目概要溶质载体 (SLC) 和 ATP 结合盒 (ABC) 超家族中的膜转运蛋白至关重要溶质（包括药物、膳食补充剂和营养素）跨膜运动中的作用。上皮细胞极化膜上的转运蛋白，例如胎盘、乳腺和肠道或极化内皮细胞，例如血脑屏障 (BBB) 的内皮细胞，在其底物的跨细胞通量。我们对 SLC 和 ABC 运输商的理解存在重大差距存在的原因是许多转运蛋白尚未被系统表征并且底物的范围未知或了解甚少。为了响应 RFA-HD-23-003，我们建议建立一个多学科运输机构加州大学旧金山分校 (TECUCSF) 阐明中心汇集了专家转运蛋白生物学和药理学、膜转运蛋白结构生物学、计算科学家生物学和化学信息学。 TECUCSF 的总体目标是阐明配体特异性、蛋白质人类 BBB 中 SLC 和 ABC 转运蛋白的结构和细胞定位。根据初步研究根据我们的全球蛋白质组分析，我们将优先列出 30 个高度表达且研究不足/孤儿 SLC 的列表和人类 BBB 中的 ABC 转运蛋白 (BBB-30)。这 30 个运输商的关键知识差距将是通过在三个目标下进行的研究来解决这一问题。对于目标 1，我们将进行神经活性物质的计算机对接使用实验确定的结构或基于以下结构的药物、营养素和膳食补充剂计算 BBB-30 中转运蛋白的结构模型。对于每个运输商，我们将创建表达转运蛋白的重组细胞系和功能测定以验证计算机结果，这将导致发现了未被研究的转运蛋白和孤儿转运蛋白的配体。对于目标 2，我们将使用冷冻电镜确定 BBB-30 中选定的转运蛋白的结构，这些转运蛋白没有可用的结构。最后，对于目标 3，我们将使用可用的抗体在人内皮细胞模型中进行 BBB-30 的定位将进行通量研究以确认定位。支持我们的研究并加强研究转运蛋白阐明网络（TEN），将建立三个核心：TECUCSF信息学和建模核心； TECUCSF结构核心；和 TECUCSF 功能核心。这些核心将为中提出的研究提供支持这三个目标中的每一个目标，以及十个研究人员的目标。 TECUCSF 将与其他 TEC 合作十项创造知识和资源，通过以下方式与更广泛的研究界共享决心增进我们对营养素、药物和膳食补充剂成分跨界运输的理解 BBB。