Deciphering biased agonistic activation of mu-opioid receptor by novel optogenetic hydrogen peroxide sensor
新型光遗传学过氧化氢传感器破译μ阿片受体的偏向激动激活
基本信息
- 批准号:10604662
- 负责人:
- 金额:$ 4.16万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2023
- 资助国家:美国
- 起止时间:2023-03-01 至 2025-02-28
- 项目状态:未结题
- 来源:
- 关键词:AcuteAddictive BehaviorAddressAnalgesicsAnimalsAnxietyArrestinsBehaviorBinding ProteinsBrainCellsChronicClinicCodeineComplexCouplingDependenceDetectionDirected Molecular EvolutionDiseaseDoseDrug AddictionDrug DesignDrug ExposureDrug ToleranceDrug Withdrawal SymptomsEmbryoEmotionsEngineeringEnhancersEnterobacteria phage P1 Cre recombinaseEpidemicEquipmentExhibitsFaceFentanylFundingFutureG Protein-Coupled Receptor SignalingGTP-Binding ProteinsGoalsGreen Fluorescent ProteinsHealthHomologous GeneHumanHydrogen PeroxideIn VitroInjectionsKineticsLigandsLinkMAPK8 geneMachine LearningMeasuresMediatingMolecularMolecular TargetMonitorMorphineMusNADPH OxidaseNational Institute of Drug AbuseNeurobiologyNeurodegenerative DisordersNeuronal PlasticityNeuronsOpioidOpioid AnalgesicsOpioid ReceptorOpioid agonistOutputPainPain managementPathologicPathway interactionsPerformancePersonsPharmaceutical PreparationsPharmacologyPhysiological AdaptationProblem SolvingProductionPropertyProtein Activation PathwayProtein EngineeringProteinsPublic HealthReactionReaction TimeReactive Oxygen SpeciesReceptor ActivationReporterResearchRodent ModelSignal PathwaySignal TransductionSiteSliceSourceSpecificityStressStructureSystemTechnologyTimeTissuesToxic effectUnited States Dept. of Health and Human ServicesUniversitiesValidationVentilatory DepressionVentral Tegmental AreaVirusWashingtonaddictionanimal tissuebasebrain tissuecell typeclinically relevantdesensitizationdesigndisease phenotypedrug developmentdrug withdrawalfluorescence imaginghigh throughput screeningimprovedin vivoinnovationinsightkappa opioid receptorskidney cellmotivated behaviormu opioid receptorsneurotransmissionnovelopioid epidemicopioid exposureopioid overdoseopioid useoptogeneticsoverdose deathperoxiredoxinpreventprototypequantumreceptorresponsesensorside effectspatiotemporaltargeted imagingtool
项目摘要
ABSTRACT: In 2017 the U.S. Department of Health and Human Services declared the ongoing opioid epidemic
a public health crisis and more than 100,000 people died due to opioid overdose in 2021. A critical part of the
solution is to understand the fundamental reaction and adaptation of brain circuits to stimulation by opioids. For
example, the desensitization of opioid receptors is a critical problem in pain management because it requires
increasing doses of analgesic compounds, which could contribute to developing a drug addiction. Recently, it
has been shown that the activation of mu and kappa opioid receptors in neurons causes the production of
reactive oxygen species (ROS) through a pathway involving NADPH oxidase and c-Jun N-terminal kinase.
Therefore, this distinct response, downstream from the receptor, could be utilized to detect specific opioid
receptor activation and modulation. However, we currently lack sensitive fluorescent sensors, which would allow
us to directly monitor pathways downstream from mu-opioid receptor (MOR) activation in real-time. Current
limitations of contemporary sensors are slow response time, low specificity, low signal output, low brightness,
and toxicity. My goal is to develop a genetically encoded sensor protein that detects ROS levels at
endogenous levels with response time and signal amplitudes that will enable monitoring of neuronal
systems upon MOR activation in brain tissue. I have developed a novel fluorescent ROS sensor with
significantly improved signaling amplitude, sensitivity, and response kinetics compared to previous sensors. I
used a newly identified insertion site on OxyR, a bacterial hydrogen peroxide binding protein, that putatively
improved allosteric coupling to the fluorescent reporter domain. We will increase the fidelity of this tool with new
green fluorescent protein (AausFP1) that exhibits exceptional quantum yield and brightness. I will optimize a
new ROS sensor at an unprecedented rate through a multifaceted approach that combines rational,
computational, and evolutionary protein engineering. My objective is to express this novel tool in MOR positive
neurons and to link ROS signals to MOR activity pharmacologically. I hypothesize that ROS signals in MOR
neurons will increase under morphine but not fentanyl through a pathway including c-Jun N-terminal Kinase,
Peroxiredoxin 6 and NADPH oxidase. Furthermore, I hypothesize that we will observe a decrease in ROS
transients under repeated drug exposure reflecting the desensitization of MORs. At the end, I will have a novel
and highly specific sensor for monitoring opioid receptor activity and adaptivity. My proposal is significant
because, for the first time, we will be able to monitor the adaptation of this clinically relevant signaling pathway
to opioid exposure. My approach is innovative because it combines novel protein engineering and monitoring
of opioid-triggered signals to dissect a difficult-to-access neuronal signaling pathway. Furthermore, this approach
could be broadly applied in future studies to monitor the activity of opioid receptors during drug exposure and
link the subsequent changes in neuronal signaling and plasticity to motivated behaviors, or analgesic tolerance.
摘要:2017年,美国卫生与公众服务部宣布阿片类药物持续流行
项目成果
期刊论文数量(0)
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科研奖励数量(0)
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专利数量(0)
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Justin Lee其他文献
Justin Lee的其他文献
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{{ truncateString('Justin Lee', 18)}}的其他基金
Ultrasound-Controlled Immunotherapy for Targeted Treatment of Solid Tumors
超声控制免疫疗法用于实体瘤的靶向治疗
- 批准号:
10399420 - 财政年份:2021
- 资助金额:
$ 4.16万 - 项目类别:
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