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A 3D Model of Human Brain Development for Studying Gene/Environment Interactions

用于研究基因/环境相互作用的人脑发育 3D 模型

基本信息

批准号：
8667546
负责人：
Thomas Hartung
金额：
$ 7.17万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-24 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8667546
关键词：
Address Adherent Culture Affect Animal Testing Apoptosis Applications Grants Astrocytes Biochemical Biological Assay Brain Calcium Cell Communication Cell Culture Techniques Cell Line Cells Chemicals Chromosome abnormality Coculture Techniques Complex Development Disease Doctor of Philosophy Down Syndrome Epigenetic Process Fluorescent Probes Gene Expression Genetic Genetic Polymorphism Glutamates Goals Human Immunohistochemistry Individual Inflammation Intervention Measures Mitochondria Modeling Modification Monitor Morphology Mutation Neurodevelopmental Disorder Neuronal Differentiation Neurons Neurotoxins Organism Oxidative Stress Pathology Patients Pharmaceutical Preparations Physiology Predictive Value Predisposition Process Psychological reinforcement Rattus Reactive Oxygen Species Reporter Genes Research Reverse Transcriptase Polymerase Chain Reaction Rodent Signal Pathway Staging Synapses Synaptic plasticity Syndrome Testing Time Toxicity Tests Tuberous sclerosis protein complex axon growth base brain cell cell type developmental neurotoxicity drug testing experience gene environment interaction human stem cells in vitro Model induced pluripotent stem cell migration myelination neurodevelopment neurotransmitter biosynthesis relating to nervous system response synaptogenesis three-dimensional modeling tool toxicant

项目摘要

DESCRIPTION (provided by applicant): A 3 D model will be developed in aggregate cultures to conduct developmental neurotoxicity testing (DNT). The model will capture the complex morphological and biochemical interactions between astrocytes and neurons that take place during development. Astrocytes participate in neuronal differentiation by providing a matrix for axonal growth and dendritic arborization. They participate in synapse formation and monitor synaptic activity through their processes that ensheath synapses. Astrocytes remove glutamate from the synapse, they respond to glutamate and ATP through a calcium response, and they detoxify reactive oxygen species generated by the active mitochondria and by neurotransmitter biosynthesis. Aggregate cultures were found to promote neuronal differentiation, synapse formation, and myelination in brains cells derived from rodents and humans. In the specific aims, neuronal differentiation, synapse formation, synaptic plasticity, and responsiveness to substances will be compared between co-cultures of astrocytes and neurons in aggregate and monolayer cultures. Astrocytes and neurons will be developed from human induced pluripotent stem cell lines (iPSC). Synapse formation will be assessed morphologically. Astrocyte participation in synapse formation will be determined morphologically by measuring the juxtaposition of astrocyte processes with synapses. Functional assessment of astrocyte participation will be accomplished with genetically driven calcium sensitive fluorescent probes. Similarly, the responses of neurons and astrocytes to different substances that increase oxidative stress and inflammation will be measured with biochemical assays and reporter gene constructs containing fluorescent probes. Because genetic variability will influence responses to drugs and other substances, the DNT model will also be developed with iPSC from different genetic backgrounds. To demonstrate the feasibility that genetic background is important in DNT, iPSC from individuals with Down's Syndrome and Tuberous Sclerosis Complex will be tested. In both syndromes, the genetic modification increases sensitivity to substances that generate oxidative stress. Several DNT substances will be examined in aggregate cultures of neurons and astrocytes induced from iPSC that are derived from both syndromes. Overall, the model developed in this proposal will have the potential to assess DNT in human neurons and astrocyte derived from individuals with different genetic backgrounds.

描述（由申请方提供）：将在聚合物培养物中开发3D模型，以进行发育神经毒性试验（DNT）。该模型将捕捉在发育过程中发生的星形胶质细胞和神经元之间复杂的形态和生化相互作用。星形胶质细胞通过为轴突生长和树突分支提供基质参与神经元分化。它们参与突触的形成，并通过包裹突触的过程监测突触活动。星形胶质细胞从突触中清除谷氨酸，它们通过钙反应对谷氨酸和ATP做出反应，并且它们对活性线粒体和神经递质生物合成产生的活性氧进行解毒。聚集体培养物被发现可以促进啮齿动物和人类脑细胞的神经元分化、突触形成和髓鞘形成。在具体的目标，神经元分化，突触形成，突触可塑性，和物质的反应性之间的共培养物的星形胶质细胞和神经元的聚集体和单层培养物进行比较。星形胶质细胞和神经元将从人诱导多能干细胞系（iPSC）发育。将从形态学上评估突触形成。星形胶质细胞参与突触形成将通过测量星形胶质细胞过程与突触的并置在形态学上来确定。星形胶质细胞参与的功能评估将用遗传驱动的钙敏感荧光探针完成。类似地，神经元和星形胶质细胞对增加氧化应激和炎症的不同物质的反应将用生化测定和含有荧光探针的报告基因构建体来测量。由于遗传变异性会影响对药物和其他物质的反应，因此DNT模型也将使用来自不同遗传背景的iPSC进行开发。为了证明遗传背景在DNT中重要的可行性，将测试来自患有唐氏综合征和多发性硬化症的个体的iPSC。在这两种综合征中，基因修饰增加了对产生氧化应激的物质的敏感性。几种DNT物质将在由两种综合征衍生的iPSC诱导的神经元和星形胶质细胞的聚集培养物中进行检查。总的来说，本提案中开发的模型将有可能评估来自不同遗传背景个体的人类神经元和星形胶质细胞中的DNT。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

State-of-the-art of 3D cultures (organs-on-a-chip) in safety testing and pathophysiology.

DOI：
10.14573/altex.1406111
发表时间：
2014
期刊：
ALTEX
影响因子：
0
作者：
Alépée N;Bahinski A;Daneshian M;De Wever B;Fritsche E;Goldberg A;Hansmann J;Hartung T;Haycock J;Hogberg H;Hoelting L;Kelm JM;Kadereit S;McVey E;Landsiedel R;Leist M;Lübberstedt M;Noor F;Pellevoisin C;Petersohn D;Pfannenbecker U;Reisinger K;Ramirez T;Rothen-Rutishauser B;Schäfer-Korting M;Zeilinger K;Zurich MG
通讯作者：
Zurich MG

Human IPSC 3D brain model as a tool to study chemical-induced dopaminergic neuronal toxicity.

人IPSC 3D脑模型是研究化学诱导的多巴胺能神经元毒性的工具。

DOI：
10.1016/j.nbd.2022.105719
发表时间：
2022-07
期刊：
NEUROBIOLOGY OF DISEASE
影响因子：
6.1
作者：
Pamies, David;Wiersma, Daphne;Katt, Moriah E.;Zhao, Liang;Burtscher, Johannes;Harris, Georgina;Smirnova, Lena;Searson, Peter C.;Hartung, Thomas;Hogberg, Helena T.
通讯作者：
Hogberg, Helena T.

Rotenone exerts developmental neurotoxicity in a human brain spheroid model.

DOI：
10.1016/j.taap.2018.02.003
发表时间：
2018-09-01
期刊：
Toxicology and applied pharmacology
影响因子：
3.8
作者：
Pamies D;Block K;Lau P;Gribaldo L;Pardo CA;Barreras P;Smirnova L;Wiersma D;Zhao L;Harris G;Hartung T;Hogberg HT
通讯作者：
Hogberg HT

21st Century Cell Culture for 21st Century Toxicology.