Rapid development of in vivo models for countermeasure discovery: organophosphate

描述（由申请人提供）： 化学对策发现的一个主要问题是可能出现新的代理人，没有已知的解毒剂或暴露后治疗。经典的反措施往往是偶然发现的，或者是经过多年的发展。随着新型化学威胁的出现（以新型化学战剂或环境污染物的形式出现），我们能够理解和应对每一种威胁的速度将决定其社会影响的大小。一个有前途的方法，快速识别和操纵的分子途径的反应，任何化学威胁是使用基于表型的化学筛选。可以开发特定毒物导致细胞或整个生物体中可再现表型的模型。通过对这些模型进行高通量筛选（HTS），可以鉴定出通过多种新机制逆转表型的小分子。通过这些筛选发现的小分子将是用于新对策的优秀先导化合物和用于解剖毒性途径的有力工具，这反过来可能指向额外的对策目标。在这个应用中，我们概述了一个过程中快速发展的体内有机磷毒性模型和它们的使用发现新的有机磷对策。虽然我们预计拟议的项目将直接导致新的有机磷对策的发展，我们也希望它作为一个模型，快速对策发展，可广泛应用于其他现有的和新兴的化学威胁。具体而言，我们建议：目标1。建立并验证斑马鱼体内有机磷毒性的检测方法。我们将寻求确定生理反应，是已知的人类有机磷反应的替代品，并可以在体内筛选高通量的规模。目标2.为了在多孔板中扩展这些用于自动化高通量筛选的新测定。我们将开发可以在96孔格式中自动进行的有机磷毒性测定。目标3.确定新的化合物，抵消先前的有机磷暴露的影响。高通量筛选将确定有助于从有机磷暴露中恢复的化合物

DESCRIPTION (provided by applicant): A major problem in chemical countermeasure discovery is the potential emergence of novel agents for which there are no known antidotes or post exposure therapies. Classic countermeasures have often been discovered only serendipitously or have taken years to develop. As novel chemical threats emerge (in the form of novel chemical warfare agents or environmental pollutants), the speed with which we are able to understand and counteract each threat will determine the magnitude of its societal impact. One promising approach for rapidly identifying and manipulating the molecular pathways underlying the response to any chemical threat is the use of phenotype-based chemical screens. Models can be developed in which specific toxicants result in reproducible phenotypes in cells or whole organisms. By subjecting these models to high- throughput screening (HTS), small molecules could be identified that reverse the phenotype through a variety of novel mechanisms. Small molecules discovered by these screens would be excellent lead compounds for novel countermeasures and powerful tools for dissecting toxicity pathways, which may in turn point to additional countermeasure targets. In this application, we outline a process for rapid development of in vivo organophosphate toxicity models and their use in discovery of novel organophosphate countermeasures. Although we expect the proposed project will lead directly to development of novel organophosphate countermeasures, we also expect it to serve as a model for rapid countermeasure development that can be applied broadly to other existing and emerging chemical threats. Specifically, we propose: Aim 1. To develop and validate in vivo assays for organophosphate toxicity in the zebrafish. We will seek to identify physiological responses that are surrogates for known human responses to organophosphates and that can be scaled for high throughput in vivo screening. Aim 2. To scale these novel assays for automated high-throughput screening in multiwell plates. We will develop assays for organophosphate toxicity that can be performed automatically in 96-well format. Aim 3. To identify novel compounds that counteract the effects of prior organophosphate exposure. High-throughput screening will identify compounds that facilitate recovery from organophosphate exposure