Developmental neurotoxicity pathways for high throughput testing by metabolomics

通过代谢组学进行高通量测试的发育神经毒性途径

• 批准号: 8334338
• 负责人: Thomas Hartung
• 金额: $ 40.52万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-21 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8334338
• 关键词: Developmental; neurotoxicity; pathways; throughput; testing

PROJECT SUMMARY There exists critical concern that exposures to drugs and chemicals during early life contribute to the increasing incidence of neurodevelopmental disorders, such as lowered IQ, learning disabilities, autism and attention deficit and hyperactivity disorder (ADHD). The developing brain is more susceptible to substance-induced injury compared to the adult brain due to the complex developmental processes, the absence of a functional blood/brain-barrier and a diminished ability to detoxify chemicals. Demanding animal tests have been devised, but because of low frequencies of hazardous substances and manifestations, as well as complex underlying mechanisms, limitations of current approaches are enormous. The area is therefore of key interest for new approaches, as outlined in the NRC vision document for a toxicology in the 21st century and the critical path initiative. The applicants have steering such a paradigm change. As a pilot reproductive toxicity concern, we propose to address the developmental neurotoxicity area, since this is a stand-alone health issue not adequately covered by current testing strategies. It represents a key element in the current transition from the two-generation study to an extended one-generation study and profits from an explosion of knowledge from basic science. In a 6-year process with critical involvement of the applicants, including three international conferences and a workshop co-organized, promising models and prototypic test substances have been identified. Extending our recent metabolomics and genomics approaches, a systems toxicology approach shall be applied now, in order to identify and validate critical pathways of toxicity (PoT). PoT-specific reporter gene models shall then be established allowing higher test throughput. We will make use of our rat primary three- dimensional organotypic in vitro model, i.e. aggregating brain cell cultures, which was shown to closely reproduce the in vivo situation of the CNS and neurotoxic mechanisms. Moreover, neurodevelopment processes have been well characterized, making the model relevant for DNT studies. Cell cultures will be exposed to DNT reference compounds during development and the expression levels of cell-specific genes will be quantified by real-time PCR. Furthermore, low molecular weight metabolites relevant for neurodevelopment will be quantified by mass spectrometry based metabolomics. Results shall give insight into the specific cell types targeted and the neurodevelopmental alterations induced. The creation of reporter gene assays in murine embryonic stem cells shall make PoT-specific testing with higher throughput possible. This will advance our understanding of DNT and test capabilities.

项目总结 存在着严重的担忧，即在生命早期接触药物和化学物质会导致 神经发育障碍的发病率增加，如智商降低，学习能力下降 残疾、自闭症、注意力缺陷和多动障碍(ADHD)。发育中的大脑 与成人大脑相比，由于复杂的大脑，更容易受到物质诱导的损伤 发育过程中，缺乏功能性血/脑屏障和 使化学物质解毒的能力。要求苛刻的动物试验已经被设计出来，但因为低 危险物质和表现形式的频率以及复杂的潜在因素 机制方面，目前方法的局限性是巨大的。因此，该地区至关重要。 对新方法的兴趣，如NRC 21世纪毒理学愿景文件中概述的那样 世纪和关键路径倡议。申请者引领了这样一种范式的转变。 作为一个试验性生殖毒性问题，我们建议解决发育神经毒性。 这是一个独立的健康问题，目前的测试没有充分涵盖这一问题 战略。它代表了当前从两代人的研究向 延续一代人的研究，并从基础知识的爆炸性增长中获益 科学。在申请者关键参与的6年过程中，包括3名 共同举办的国际会议和研讨会，前景看好的模型和原型测试 物质已被鉴定出来。扩展我们最新的代谢组学和基因组学 方法，现在应应用系统毒理学方法，以确定和 验证毒性的关键途径(POT)。然后，特定于POT的报告基因模型应 已建立，允许更高的测试吞吐量。我们将利用我们的老鼠小学三年级- 三维器官型体外模型，即聚集脑细胞培养，表明 紧密复制中枢神经系统的体内情况和神经毒性机制。此外， 神经发育过程已经被很好地描述，这使得该模型与DNT相关 学习。细胞培养在发育过程中将暴露于DNT参比化合物中， 细胞特异性基因的表达水平将通过实时聚合酶链式反应进行量化。此外，Low 与神经发育相关的分子质量代谢物将按质量进行量化 以光谱为基础的代谢组学。结果应提供对目标特定细胞类型的洞察 以及神经发育的改变引起的。报告基因分析技术在中国的创造 小鼠胚胎干细胞将使POT特异性测试成为可能，并有可能获得更高的产量。 这将增进我们对DNT和测试能力的理解。

项目成果

Look back in anger - what clinical studies tell us about preclinical work.

• DOI: 10.14573/altex.2013.3.275
• 发表时间: 2013
• 期刊: ALTEX
• 作者: Hartung T

Biological and medical applications of a brain-on-a-chip.

• DOI: 10.1177/1535370214537738
• 发表时间: 2014-09
• 期刊: Experimental biology and medicine (Maywood, N.J.)
• 作者: Pamies D;Hartung T;Hogberg HT
• 通讯作者: Hogberg HT

Organophosphorus flame retardants are developmental neurotoxicants in a rat primary brainsphere in vitro model.

• DOI: 10.1007/s00204-020-02903-2
• 发表时间: 2021-01
• 期刊: Archives of toxicology
• 影响因子: 6.1
• 作者: Hogberg HT;de Cássia da Silveira E Sá R;Kleensang A;Bouhifd M;Cemiloglu Ulker O;Smirnova L;Behl M;Maertens A;Zhao L;Hartung T
• 通讯作者: Hartung T

Mechanistic validation.

机械验证。

• DOI: 10.14573/altex.2013.2.119
• 发表时间: 2013
• 期刊: ALTEX
• 作者: Hartung,Thomas;Hoffmann,Sebastian;Stephens,Martin
• 通讯作者: Stephens,Martin

Review: toxicometabolomics.

• DOI: 10.1002/jat.2874
• 发表时间: 2013-12
• 期刊: JOURNAL OF APPLIED TOXICOLOGY
• 影响因子: 3.3
• 作者: Bouhifd, Mounir;Hartung, Thomas;Hogberg, Helena T.;Kleensang, Andre;Zhao, Liang
• 通讯作者: Zhao, Liang