Functional characterization of SSRI resistant genes using C. elegans

使用线虫对 SSRI 抗性基因进行功能表征

• 批准号: 7835764
• 负责人: JI Y SZE
• 金额: $ 63.53万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-07 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7835764
• 关键词: Acetylcholine; Animal Model; Animals; Behavior; Behavioral; Behavioral Paradigm; Binding; Biochemical Markers; Biological Assay; Biological Models; Biology; C. elegans genome; Caenorhabditis elegans; Candidate Disease Gene; Cell physiology; Cells; Data; Development; Disease; Dissection; Drug Delivery Systems; Drug resistance; Epitopes; Etiology; Excision; Family; Fluoxetine; Gene Expression Profiling; Genes; Genetic; Genetic Predisposition to Disease; Genetic Screening; Genome; Glutamates; Goals; Individual; Insulin Receptor; Life; Maps; Mental disorders; Molecular; Monitor; Mutagenesis; Mutation; Neurons; Neurotransmitters; Organism; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phenotype; Physiological; Physiology; Prozac; Publishing; RNA Interference; Receptor Signaling; Regulation; Resistance; Resolution; Role; Selective Serotonin Reuptake Inhibitor; Serotonin; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism Map; Stress; Structure; Symptoms; Synapses; System; Technology; Testing; Therapeutic Effect; base; biological adaptation to stress; calmodulin-dependent protein kinase II; drug efficacy; efficacy testing; gamma-Aminobutyric Acid; genome-wide; molecular marker; mutant; neurotransmission; novel therapeutics; postsynaptic; public health relevance; receptor; research study; response; reuptake; serotonin receptor; serotonin transporter

DESCRIPTION (provided by applicant): The goal of this proposal is to elucidate the serotonin signaling pathways through genetic dissection of genes and functional pathways regulated by Selective Serotonin Reuptake Inhibitors (SSRIs). Dysregulation of the serotonergic system has been implicated in a wide-spectrum of neurodevelopmental and psychiatric disorders, and SSRIs have made a major impact on many symptoms of these diseases. SSRIs are thought to exert therapeutic effects by inhibiting the serotonin reuptake transporter (SERT), thereby increasing serotonin signaling, leading to our central hypothesis that genetic dissection of SSRIs-resistant genes will identify not only the genes directly binding the drugs, but also the genes and functional pathways regulated by the drugs, and therefore, will shed lights on the fundamental biology of serotonin signaling and its role in the diseases. Taking the advantage of the availability of serotonin-deficient C. elegans, and the unique ability to monitor gene expression and synapse structure in identified cells in the living worm, this proposal uses direct forward genetic approaches in C. elegans to identify SSRIs-resistant genes in a whole animal. This proposal is based on our preliminary data indicating that the SSRI fluoxetine (Prozac ) targets serotonin postsynaptic targets to regulate the activity of acetylcholine, GABA, and glutamate neurotransmission. In four specific aims, proposed experiments will search genome-wide for mutant worms that are resistant to fluoxetine to elucidate the genetic interactions between SSRIs, serotonin, and the other neurotransmitters. The first aim is to identify fluoxetine- resistant genes. We have established a behavioral paradigm for genetic screening for fluoxetine-resistant mutants. Using this paradigm, we have already identified 5 genes known to be involved in serotonin signaling. In addition, we have isolated 35 new fluoxetine-resistant mutants through an unbiased mutagenesis screen. We will characterize the new mutants and clone the mutant genes. The second aim is to characterize the role of new fluoxetine-resistant genes in regulating acetylcholine, GABA and glutamate neurotransmission, and to organize the drug-resistant genes in functional pathways that couple serotonin and individual neurotransmitter signaling pathways. The third aim is to characterize the role of the fluoxetine-resistant genes in serotonin downstream pathways that regulate developmental and physiological responses to environmental stresses. The fourth aim is to examine the genetic interactions between new fluoxetine-resistant genes and serotonin receptors. By testing serotonin deficient mutants, SERT-null mutants, serotonin receptor mutants, and fluoxetine resistant mutants, we will evaluate the comparability of serotonin, SSRIs and SERT on the function of serotonin receptors in regulating the synaptic activity of the other neurotransmitters, development, and stress physiology. PUBLIC HEALTH RELEVANCE: The systematic genetic dissection of SSRIs downstream targets in a model organism will reveal core components and principal functional pathways of the serotonergic system that exert conserved functions in disparate organisms and are important for the therapeutic effects of SSRIs. These functional pathways may also provide biochemical markers for subdividing a particular disease and reveal shared genetic vulnerability that might unify distinct disorders. These in turn will provide clues about the genetic basis underlying the pathogenesis and offer new windows for the discovery of novel therapeutics.

描述（由申请人提供）：该提案的目的是通过基因的遗传解剖和由选择性5-羟色胺再摄取抑制剂（SSRIS）调节的功能途径来阐明5-羟色胺信号通路。血清素能系统的失调与广泛的神经发育和精神疾病有关，SSRI对这些疾病的许多症状产生了重大影响。 SSRIs are thought to exert therapeutic effects by inhibiting the serotonin reuptake transporter (SERT), thereby increasing serotonin signaling, leading to our central hypothesis that genetic dissection of SSRIs-resistant genes will identify not only the genes directly binding the drugs, but also the genes and functional pathways regulated by the drugs, and therefore, will shed lights on the fundamental biology of serotonin信号及其在疾病中的作用。利用缺乏5-羟色胺的秀丽隐杆线虫的可用性，以及监测活蠕虫中鉴定出的细胞中基因表达和突触结构的独特能力，该提案使用秀丽隐杆线虫中的直接前向遗传方法来识别整个动物中对SSRIS耐药基因。该建议基于我们的初步数据，表明SSRI氟西汀（Prozac）靶向5-羟色胺后突触靶标，以调节乙酰胆碱，GABA和谷氨酸神经传递的活性。在四个特定目标中，提出的实验将在全基因组中搜索对氟西汀具有抗性的突变蠕虫，以阐明SSRIS，5-羟色胺和其他神经递质之间的遗传相互作用。第一个目的是鉴定抗氟西汀 - 耐药基因。我们已经建立了一种行为范例，用于筛选抗氟西汀的突变体的基因筛查。使用此范式，我们已经确定了5个已知参与5-羟色胺信号传导的基因。此外，我们通过无偏的诱变筛选分离了35个新的抗氟西汀耐药突变体。我们将表征新的突变体并克隆突变基因。第二个目的是表征新的抗氟西汀基因在调节乙酰胆碱，GABA和谷氨酸神经传递中的作用，并在功能途径中组织药物耐药基因，这些途径将伴随5-羟色胺和单个神经递质信号传导途径。第三个目的是表征氟西汀耐药基因在调节环境应激的发育和生理反应的血清素下游途径中的作用。第四个目的是检查新的耐氟西汀基因和5-羟色胺受体之间的遗传相互作用。通过测试5-羟色胺缺乏突变体，Sert-Null突变体，5-羟色胺受体突变体和抗氟西汀抗性突变体，我们将评估5-羟色胺，SSRIS和SERT的可比性在调节其他神经交易者突触活性中的5-羟色胺，SSRIS和SERT对其他神经递质的突触活性。公共卫生相关性：模型生物体中SSRIS下游靶标的系统遗传解剖将揭示血清素能系统的核心成分和主要功能途径，这些核能系统在不同的生物体中发挥了保守作用，并且对SSRIS的治疗作用很重要。这些功能途径也可能提供生化标志物来细化特定疾病，并揭示共同的遗传脆弱性，可能统一不同的疾病。这些反过来将提供有关发病机理的遗传基础的线索，并为发现新型治疗学提供新的窗口。

项目成果

Disruption of Transient SERT Expression in Thalamic Glutamatergic Neurons Alters Trajectory of Postnatal Interneuron Development in the Mouse Cortex.

丘脑谷氨酸能神经元瞬时 SERT 表达的破坏会改变小鼠皮层出生后中间神经元发育的轨迹。

• DOI: 10.1093/cercor/bhz191
• 发表时间: 2020
• 期刊: Cerebral cortex (New York, N.Y. : 1991)
• 作者: DeGregorio,Roberto;Chen,Xiaoning;Petit,EmilieI;Dobrenis,Kostantin;Sze,JiYing
• 通讯作者: Sze,JiYing

Spatiotemporal SERT expression in cortical map development.

• DOI: 10.1016/j.neuint.2016.05.010
• 发表时间: 2016-09
• 期刊: Neurochemistry international
• 影响因子: 4.2
• 作者: Chen X;Petit EI;Dobrenis K;Sze JY
• 通讯作者: Sze JY