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: 7508941
• 负责人: JI Y SZE
• 金额: $ 61.84万
• 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 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7508941
• 关键词: Acetylcholine; Affect; Amendment; Americas; Animal Model; Animals; Antidepressive Agents; Antioxidants; Applications Grants; Autistic Disorder; Award; Behavior; Behavioral; Behavioral Paradigm; Binding; Biochemical; Biochemical Markers; Biological Assay; Biology; Budgets; C. elegans genome; Caenorhabditis elegans; Candidate Disease Gene; Cells; Chemicals; Chromosome Mapping; Citalopram; Classification; Cloning; Collaborations; Data; Development; Diabetes Mellitus; Disease; Dissection; Drug effect disorder; Drug resistance; Fluoxetine; Funding; Future; Gene Expression Profiling; Gene Targeting; Genes; Genetic; Genetic Predisposition to Disease; Genetic Screening; Glutamates; Goals; Grant; Homeostasis; Imipramine; Individual; Laboratories; Life; Maps; Medicine; Mental disorders; Metabolic; Mitochondria; Molecular; Motivation; Mus; Mutagenesis; Mutant Strains Mice; Nervous system structure; Neurons; Neurotransmitters; Organism; Oxidative Stress; Paroxetine; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Physiological; Physiology; Postdoctoral Fellow; Process; Proteins; Prozac; RNA Interference; Receptor Signaling; Recovery; Regulation; Reporter; Resistance; Role; Selective Serotonin Reuptake Inhibitor; Serotonin; Serotonin Agents; Sertraline; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism Map; Stress; Structure; Symptoms; Synapses; Synaptic Transmission; System; Testing; Therapeutic Effect; Time; Translating; Translations; Universities; Wing; Work; base; biological adaptation to stress; college; design; endophenotype; experience; gamma-Aminobutyric Acid; genetic analysis; genome-wide; graduate student; inhibitor/antagonist; insight; mutant; neurotransmission; novel; novel therapeutics; postsynaptic; public health relevance; receptor; research study; response; reuptake; serotonin receptor; success

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）：本提案的目标是通过对选择性5-羟色胺再摄取抑制剂（SSRI）调节的基因和功能途径进行遗传解剖，阐明5-羟色胺信号传导途径。多巴胺能系统的失调与广泛的神经发育和精神疾病有关，SSRIs对这些疾病的许多症状产生了重大影响。SSRIs被认为通过抑制5-羟色胺再摄取转运蛋白（SERT）发挥治疗作用，从而增加5-羟色胺信号传导，导致我们的中心假设，即SSRIs抗性基因的遗传解剖不仅可以识别直接结合药物的基因，而且还可以识别受药物调节的基因和功能途径，因此，将揭示血清素信号的基本生物学及其在疾病中的作用。利用低维C. elegans，以及在活蠕虫中识别细胞中监测基因表达和突触结构的独特能力，该提议在C. elegans来鉴定整个动物的SSRIs抗性基因。这个建议是基于我们的初步数据表明，SSRI氟西汀（百忧解）的目标是5-羟色胺突触后靶点，以调节乙酰胆碱，GABA和谷氨酸神经传递的活性。在四个具体目标中，拟议的实验将在全基因组范围内搜索对氟西汀具有抗性的突变蠕虫，以阐明SSRIs，血清素和其他神经递质之间的遗传相互作用。第一个目标是确定氟西汀耐药基因。我们已经建立了一个行为范式的氟西汀耐药突变体的遗传筛选。使用这种模式，我们已经确定了5个已知参与血清素信号传导的基因。此外，我们已经分离出35个新的氟西汀耐药突变体，通过无偏诱变筛选。我们将描述新的突变体并克隆突变基因。第二个目的是表征新的氟西汀耐药基因在调节乙酰胆碱、GABA和谷氨酸神经传递中的作用，并将耐药基因组织在耦合5-羟色胺和个体神经递质信号传导通路的功能通路中。第三个目的是描述氟西汀耐药基因在5-羟色胺下游通路中的作用，该通路调节对环境压力的发育和生理反应。第四个目标是研究新的氟西汀耐药基因和5-羟色胺受体之间的遗传相互作用。通过检测5-羟色胺缺乏突变体、SERT无效突变体、5-羟色胺受体突变体和氟西汀耐药突变体，我们将评估5-羟色胺、SSRIs和SERT对5-羟色胺受体在调节其他神经递质的突触活性、发育和应激生理学方面的功能的可比性。公共卫生相关性：在模式生物体中对SSRI下游靶点的系统性遗传解剖将揭示在不同生物体中发挥保守功能并且对SSRI的治疗作用重要的β-羟色胺能系统的核心组分和主要功能途径。这些功能通路也可以提供生化标记，用于细分特定疾病，并揭示可能统一不同疾病的共同遗传脆弱性。这些反过来将为发病机制的遗传基础提供线索，并为发现新的治疗方法提供新的窗口。