Identity, mechanisms and early life impacts of transporter interfering compounds

转运蛋白干扰化合物的特性、机制和早期生命影响

基本信息

批准号：
10179393
负责人：
GEOFFREY A CHANG
金额：
$ 55.86万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10179393
关键词：
ABCB1 gene ABCC1 gene ABCG2 gene Address Affect Affinity Animal Model Animals Antibodies Binding Biochemical Biological Assay Biological Models Biophysics Carrier Proteins Cell Differentiation process Cells Cellular Assay Chemical Exposure Chemicals Clustered Regularly Interspaced Short Palindromic Repeats Complex Cryoelectron Microscopy Data Development Disease Elderly Embryo Embryonic Development Exposure to Family Fishes Functional disorder Genetic Germ Lines Goals Growth Health Human Image In Vitro Knock-out Laboratory Molecular Evolution Life Ligands Liquid substance Maternal-fetal medicine Measures Modeling Molecular Molecular Structure Molecular Target Mus Performance Pharmaceutical Preparations Predisposition Program Development Regulation Reproductive Health Research Research Personnel Resistance Resolution Risk Role Sea Urchins Signal Transduction Somatic Cell Structure Structure of primordial sex cell System Technology Teratogens Testing Time Toxicant exposure Translating Umbilical Cord Blood Ursidae Family Uterus X-Ray Crystallography Xenobiotics Zebrafish data modeling early life exposure environmental chemical experimental study exposed human population human embryonic stem cell inhibitor/antagonist innovation insight nanobodies pollutant pollutant interaction prenatal exposure reproductive reproductive fitness reproductive system disorder toxicant transcriptome

项目摘要

PROJECT SUMMARY Prenatal exposures to environmental chemicals have been shown to cause adverse later life health effects, often involving disorders of reproductive dysfunction. The overall goal of this research is to understand the mechanisms governing accumulation of environmental chemicals in the embryo, so that we can predict and mitigate the negative effects of these exposures. In this proposal, we address two key questions with regard to xenobiotic accumulation in the embryo, with a specific focus on the role of xenobiotic transporters during primordial germ cell (PGC) formation. First, we ask how the program of development leads to changes in xenobiotic transporter expression, and thus generates windows of susceptibility or resistance to xenobiotic accumulation. Second, we ask how real-world chemical mixtures, containing both substrates and inhibitors of transporters, impact the efficacy of this conserved, protective system. Aim 1 uses a powerful in vitro molecular evolution technology to rapidly evolve, validate, and use antibody-like binders called nanobodies to characterize xenobiotic transporter proteins in human PGC-like cells (PGLCs) and in model organism embryos (sea urchin and zebrafish). Aim 2 applies biochemical and cellular approaches to determine relevant environmental ligands of human and model system xenobiotic transporters, and takes advantage of a powerful molecular structure determination pipeline to dissect the molecular mechanisms of these interactions. Aim 3 uses models and molecular targets from Aims 1 and 2 to test the hypothesis that PGCs are vulnerable to the interfering effects of environmental chemicals on the transporter defense system, and that disruption of this system leads to decreased reproductive fitness after xenobiotic challenge. This results will provide new insights into how environmental and developmental factors act in combination to govern the susceptibility of the nascent embryonic germ line to teratogens.

项目概要产前接触环境化学物质已被证明会对以后的健康产生不利影响，常涉及生殖功能障碍疾病。这项研究的总体目标是了解控制胚胎中环境化学物质积累的机制，以便我们可以预测和减轻这些风险的负面影响。在本提案中，我们解决了两个关键问题：胚胎中的异生素积累，特别关注异生素转运蛋白在胚胎发育过程中的作用原始生殖细胞（PGC）的形成。首先，我们要问发展计划如何导致变化异生物质转运蛋白表达，从而产生对异生物质的敏感性或抗性窗口积累。其次，我们询问现实世界中含有底物和抑制剂的化学混合物如何转运蛋白，影响这个保守的保护系统的功效。 Aim 1 使用强大的体外分子快速进化、验证和使用称为纳米抗体的抗体样结合物的进化技术表征人类 PGC 样细胞 (PGLC) 和模型生物胚胎中的异生物质转运蛋白（海胆和斑马鱼）。目标 2 应用生化和细胞方法来确定相关的人类和模型系统外源转运蛋白的环境配体，并利用强大的优势分子结构测定管道来剖析这些相互作用的分子机制。目标 3 使用目标 1 和 2 中的模型和分子靶标来检验 PGC 易受环境化学物质对转运蛋白防御系统的干扰作用，以及对该系统的破坏系统导致外源物质挑战后生殖适应性下降。这一结果将提供新的深入了解环境和发育因素如何共同作用来控制易感性新生胚胎生殖系致畸。