Genetic Variation of Ultra-Potent Synthetic Opioid Sensitivity in Mice

小鼠超强合成阿片类药物敏感性的遗传变异

基本信息

批准号：
10743432
负责人：
Jason A Bubier
金额：
$ 45.61万
依托单位：
JACKSON LABORATORY
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：
2023-07-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10743432
关键词：
Acute Address Affect African American population Apnea Automobile Driving Biological Blood Blood gas Brain Stem Candidate Disease Gene Carbon Dioxide Cardiovascular system Cations Cause of Death Cells Central Nervous System Cessation of life Chest wall structure Clustered Regularly Interspaced Short Palindromic Repeats Coupled Data Development Dose Epidemic Exhibits Fentanyl Foundations Gene Expression Genes Genetic Genetic Variation Genetic study Genotype Goals Human Inbred Strains Mice Individual Differences Knock-out Lethal Dose 50 Lung Measures Molecular Monitor Morphine Moscow Mouse Strains Mus Naloxone Opioid Opioid Analgesics Opioid Antagonist Opioid Receptor Overdose Oxygen Pathway interactions Patients Peripheral Nervous System Phenotype Physiological Plethysmography Population Recovery Respiratory Mechanics Signal Transduction Street Drugs Testing Therapeutic Tidal Volume Time Tissue-Specific Gene Expression Toxic effect Toxicokinetics Variant Ventilatory Depression Withdrawal Wooden Chest Syndrome Work addiction aerosolized analog behavioral phenotyping carfentanil cell type cohort detection platform differential expression electric impedance genome wide association study hemodynamics individual variation innovation insight molecular phenotype neural novel opioid overdose overdose death overdose risk preBotzinger complex prescription opioid psychologic respiratory response segregation sex single nucleus RNA-sequencing synthetic opioid targeted treatment therapeutic development translational impact ventilation weapons

Acute Address Affect African American population Apnea Automobile Driving Biological Blood Blood gas Brain Stem Candidate Disease Gene Carbon Dioxide Cardiovascular system Cations Cause of Death Cells Central Nervous System Cessation of life Chest wall structure Clustered Regularly Interspaced Short Palindromic Repeats Coupled Data Development Dose Epidemic Exhibits Fentanyl Foundations Gene Expression Genes Genetic Genetic Variation Genetic study Genotype Goals Human Inbred Strains Mice Individual Differences Knock-out Lethal Dose 50 Lung Measures Molecular Monitor Morphine Moscow Mouse Strains Mus Naloxone Opioid Opioid Analgesics Opioid Antagonist Opioid Receptor Overdose Oxygen Pathway interactions Patients Peripheral Nervous System Phenotype Physiological Plethysmography Population Recovery Respiratory Mechanics Signal Transduction Street Drugs Testing Therapeutic Tidal Volume Time Tissue-Specific Gene Expression Toxic effect Toxicokinetics Variant Ventilatory Depression Withdrawal Wooden Chest Syndrome Work addiction aerosolized analog behavioral phenotyping carfentanil cell type cohort detection platform differential expression electric impedance genome wide association study hemodynamics individual variation innovation insight molecular phenotype neural novel opioid overdose overdose death overdose risk preBotzinger complex prescription opioid psychologic respiratory response segregation sex single nucleus RNA-sequencing synthetic opioid targeted treatment therapeutic development translational impact ventilation weapons

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项目摘要

PROJECT SUMMARY/ABSTRACT The weaponization of Ultrapotent synthetic opioids (UPS) has made finding a novel reversal agent a priority. The current opioid response agent, Naloxone, is not as effective against UPS opioids and does not reverse one of its known effects, wooden chest syndrome. Our long-term goal is to define the biological basis of opioid overdose risk and promote the discovery of safe and effective agents that reverse fentanyl lethality. A key objective is determining the molecular mechanisms underlying individual variability to fentanyl toxicity using genetically diverse mice. In aim 1 we plan to identify genes and variants that modify the influence of Mcoln1 on acute UPS opioid toxicity. Mcoln1 was identified in a GWAS study of overdose risk, and preliminary data support a genetic knockout of Mcoln1 resulting in death more rapidly from morphine or fentanyl. We will create an additional CRISPR knockout of Mcoln1 on a more sensitive genetic background, NOD/ShiLtJ and compare it with the C57BL/6 knockout we have for response to the UPS opioid fentanyl. The LD50 will be determined for these strains using our piezoelectric respiratory depression detection system. We will also study respiratory mechanics and pulmonary and chest wall impedance in response to three doses of fentanyl. In another cohort of mice, they will be tested by plethysmography to acquire respiratory metrics such as tidal volume and minute ventilation. We will also collect arterial blood to measure blood gases of oxygen and carbon dioxide during the plethysmography session to monitor the response to fentanyl at that level. Finally, another cohort of naïve and fentanyl-treated mice will be dissected for brain stems. The pre-bötzinger complex will be identified and analyzed by single-nucleus RNA-Seq, comparing the cellular populations and differential gene expression across genotypes, sexes and treatments. In aim 2 we plan to identify the physiological, neural, and molecular mechanisms of variable fentanyl-induced toxicity and lethality among eight inbred mouse strains. These eight strains, which served as the foundation for the advanced mouse populations of the Collaborative Cross and Diversity Outbred mice, contain approximately 45 million SNPs segregating between them. We have determined that the LD50 for fentanyl varies > 150-fold across both sexes of the eight strains. As in aim 1, in aim 2 we will phenotype cohorts of mice to detect the diverse phenomena associated with UPS opioids, including Opioid- Induced Respiratory Depression (OIRD), Opioid-Induced Persistent Apnea (OIPA), Wooden Chest Syndrome (WCS), closure/collapse of the upper and cardiovascular/hemodynamic disturbances. This phenotyping will be coupled to identifying the cellular populations, through single nucleus RNA-Seq, within the brainstem pre- bötzinger region that varies across naïve and fentanyl-treated strains of both sexes of mice. The differentially expressed genes that define these populations will help us identify targets for therapeutic development associated with the different fentanyl lethality phenotypes.

项目摘要/摘要超能力合成阿片类药物（UPS）的武器化使发现一种新颖的逆转剂成为优先事项。这当前的阿片类药物反应剂纳洛酮对UPS阿片类药物不那么有效，也不会逆转一种它已知的效果，木质胸部综合征。我们的长期目标是定义阿片类药物的生物学基础过量的风险，并促进发现逆转芬太尼致死性的安全有效药物。钥匙目的是确定使用个人变异性的分子机制使用遗传多样的小鼠。在AIM 1中，我们计划确定改变McOLN1对的基因和变体急性UPS阿片类药物毒性。在GWAS的过量风险和初步数据支持中，MCOLN1被确定 MCOLN1的遗传敲除导致吗啡或芬太尼更快地导致死亡。我们将创建一个在更敏感的遗传背景上对McOLN1的其他CRIS敲除，点头/shiltj并进行比较使用C57BL/6敲除我们可以响应UPS阿片类药物芬太尼。 LD50将确定这些菌株使用我们的压电呼吸道抑郁检测系统。我们还将研究呼吸道响应三剂芬太尼的力学，肺和胸壁阻抗。在另一个队列中在小鼠中，它们将通过散布图表进行测试，以获取呼吸指标，例如潮汐量和微小通风。我们还将收集动脉血，以测量氧气和二氧化碳的血液气体杂质学会话以监测该水平的芬太尼响应。最后，另一个幼稚和芬太尼治疗的小鼠将针对大脑步骤进行剖析。将识别和分析前胞津犬综合体通过单核RNA-Seq，比较了细胞群体和差异基因表达基因型，性别和治疗方法。在AIM 2中，我们计划识别物理，神经和分子芬太尼诱导的毒性和杀伤性的八个近交小鼠菌株之间的机制。这八个菌株，是协作十字架的高级鼠标的基础多样性杂种小鼠在它们之间占有约4500万个SNP。我们已经确定芬太尼品种的LD50在八个菌株的两个性别中> 150倍。与AIM 1一样，在AIM 2中，我们将小鼠的表型人群检测与UPS阿片类药物相关的潜水现象，包括阿片类药物诱导呼吸抑郁症（OIRD），阿片类药物诱导的持续呼吸暂停（OIPA），木制胸部综合征（WCS），上部和心血管疾病的闭合/塌陷。这种表型将是通过单核RNA-seq在脑干预先识别细胞群体的结合 Bötzinger地区的两种男性两性菌株变化和芬太尼处理的菌株。定义这些人群的表达基因将有助于我们确定热发展的目标与不同的芬太尼致命表型相关。