Defining the Chemical Perturbome of Neural Development and Activity

项目摘要 数以百万计的精确细胞决策是大脑准确发育的基础。小分子可以 通过与在谱系轨迹中起关键作用的蛋白质结合来影响这些决定。他们可以调谐 通过增强或耗尽特定类型的神经元或它们的连接来促进或耗尽电路。一本化学“词典” 化合物及其对大脑的影响将是环境分类的宝贵资源 神经毒素和神经发育障碍的发现疗法。然而，我们缺乏有效的方法来 量化这些影响。 我们提出了一种多管齐下的方法来筛选化学物质的神经发育作用，整合 全生物筛选、蛋白质工程和计算机药物发现。幼虫斑马鱼将会是 我们对这些分子和新工具的试验场。首先，我们将依靠经过验证的 用于行为、大脑活动和大脑结构的高通量分析的管道。我们最近利用 开发了自由游泳动物全脑活动图谱的方法，建立在我们的屏幕上 增加神经发育疾病风险的基因突变。我们的努力将集中在自闭症上 和智力障碍：评估环境污染物表明会增加这些障碍的发生率 以及筛选临床开发中的药库，以加快治疗。第二，超越 整个动物筛选的吞吐量限制，我们将设计一个工具来转换关于 DNA读数中特定神经元类型的存在和活动。96口井中接触化合物的幼虫 车牌将与下一代测序进行多路传输。最后，药物发现的前沿是 计算性的。有数十亿个潜在的可合成分子，即使是测试一个 试验性的小百分比。我们正在开发一种新的基于计算的预测算法 将基于知识的方法与物理能量势相结合的化学-蛋白质相互作用 罗塞塔建模程序。我们正在进行的与基因相关的基础生物学研究 神经发育障碍将产生用于建模的蛋白质靶标。 随着每一个拟议的技术进步，我们将增加我们可以分析的分子的数量 数量级。我们策略的力量在于撒下一张大网来定义具有多样性的分子 操纵神经发育和活动的特定手段。我们将推进预防， 了解和治疗神经发育障碍 驱动这些疾病和那些可以逆转表型的疾病来治疗它们。

Project Summary Millions of precise cellular decisions underlie the accurate development of a brain. Small molecules can impinge on these decisions by binding to proteins with critical roles in lineage trajectories. They can tune circuits by boosting or depleting specific types of neurons or their connections. A “dictionary” of chemical compounds and their effects on the brain would be an invaluable resource for classifying environmental neurotoxins and discovering treatments for neurodevelopmental disorders. Yet, we lack efficient approaches to quantify these effects. We propose a multi-pronged approach to screen chemicals for their neurodevelopmental roles, integrating whole-organism screening, protein engineering, and computational drug discovery. The larval zebrafish will be our testing ground for these molecules and new tools. First, we will rely on improved versions of our validated pipeline for high-throughput analysis of behavior, brain activity, and brain structure. We leverage recently developed methods for whole-brain activity mapping in freely swimming animals, building upon our screens of genetic mutants for genes that increase risk of neurodevelopmental disease. Our efforts will focus on autism and intellectual disability: assessing environmental contaminants indicated to increase rates of these disorders and screening libraries of drugs in clinical development to accelerate treatments. Second, to surpass the throughput limitations of whole animal screening, we will engineer a tool to convert information about the presence and activity of specific neuron types into a DNA readout. Larvae exposed to compounds in 96-well plates will be multiplexed with next-generation sequencing. Finally, the frontier of drug discovery is computational. There are billions of potentially synthesizable molecules, and it is inconceivable to test even a small percentage experimentally. We are developing a new algorithm for computational-based prediction of chemical-protein interactions that integrates a knowledge-based approach with the physical energy potential of the Rosetta modeling program. Our ongoing studies of the basic biology of the genes linked to neurodevelopmental disorders will yield the protein targets for modeling. With each proposed technological advance, we will increase the number of molecules we can analyze by an order of magnitude. The power of our strategy lies in casting a wide net to define molecules with diverse yet specific means of manipulating neural development and activity. We will push forward the prevention, understanding, and treatment of neurodevelopmental disorders by considering both the chemicals that can drive these disorders and the ones that can reverse phenotypes to treat them.