Identity, mechanisms and early life impacts of transporter interfering compounds

项目总结 产前接触环境化学物质已被证明会对晚年健康造成不利影响， 通常涉及生殖功能障碍的障碍。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，这样我们就可以预测和 减轻这些风险暴露的负面影响。在这项提案中，我们解决了关于以下两个关键问题 异种生物在胚胎中的积累，特别关注异种转运蛋白在胚胎发育过程中的作用 原始生殖细胞(PGC)形成。首先，我们问发展计划是如何导致 异种生物转运蛋白的表达，从而产生对异种生物的易感性或抵抗力窗口 积累。其次，我们问，包含底物和抑制剂的真实世界化学混合物是如何 转运体，影响这一保守的保护系统的效力。AIM 1使用一种强大的体外分子 进化技术，以快速进化、验证和使用被称为纳米体的类似抗体的结合体来 人PGC样细胞和模型生物胚胎中异种转运蛋白的特性 (海胆和斑马鱼)。目标2应用生化和细胞方法来确定相关的 环境配体对人体和模型系统的异种生物转运蛋白，并利用强大的 分子结构测定管道，以剖析这些相互作用的分子机制。目标3 使用AIMS 1和AIMS 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体防御系统的干扰作用，以及对这种影响的破坏 系统导致异种生物攻击后生殖适合度降低。这一结果将提供新的 洞察环境和发展因素如何共同作用来控制糖尿病的易感性 从新生的胚胎生殖系到致畸剂。

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.