The role of Zhx2 in CYP2D regulation, oxycodone metabolism, and opioid addiction model behaviors.

Zhx2 在 CYP2D 调节、羟考酮代谢和阿片类药物成瘾模型行为中的作用。

基本信息

批准号：
10466122
负责人：
William Lynch
金额：
$ 4.77万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
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：
2023-08-08 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10466122
关键词：
Accounting Analgesics Behavior Behavioral Biological Assay Brain CRISPR/Cas technology CYP2D6 gene Candidate Disease Gene Cessation of life Chromosome 15 Chromosome Mapping Clinical Code Complex Corpus striatum structure Cytochrome P450 DNA Insertion Elements Data Elements Environment Enzymes Epidemic Excision Female Genes Genetic Genetic Transcription Genotype Goals Harvest Hepatic Heritability High Pressure Liquid Chromatography Hippocampus Homeobox Human Inbred BALB C Mice Inbred Mouse Learning Link Liver Liver Extract Locomotion Maps Mass Spectrum Analysis Messenger RNA Metabolism Modeling Molecular Monitor Mus Neurobiology Opiate Addiction Opioid Oral Orthologous Gene Oxycodone Oxymorphone Phenotype Plasma Predisposition Privatization Production Proteins Proteome Proteomics Public Health Quantitative Trait Loci Regulation Rewards Rodent Rodent Model Role Sample Size Self Administration Site Substance Use Disorder System Testing Therapeutic Therapeutic Uses Tissues Training Transcription Repressor United States Validation Variant Zinc Fingers Zinc deficiency addiction adeno-associated viral vector behavioral phenotyping brain tissue candidate validation causal variant conditioned place preference drug metabolism genetic manipulation genetic variant genome wide association study genome-wide in vivo individualized medicine insight interdisciplinary approach liver metabolism model organism mouse genetics novel opioid mortality opioid use disorder overexpression personalized medicine pharmacologic prescription opioid protein expression rodent genome small hairpin RNA therapeutic opioid trait transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT ABSTRACT Opioid Use Disorder (OUD) remains a major public health issue, with 2020 containing the highest number of OUD-related deaths on record. Prescription opioids contribute towards a substantial portion of OUD-related deaths, with the opioid oxycodone (OXY) among the most prescribed opioids and underlying a significant percentage of OUDs. OUD-related traits, including use and misuse, behavioral traits, and metabolism have a heritable component, yet genetic variants underlying OUD traits remain largely undiscovered. Rodent genome- wide association studies (GWAS) with quantitative trait locus (QTL) mapping can efficiently identify quantitative trait genes and mechanisms through robustly powered sample-sizes, controlled environmental conditions, and the ability to perform direct genetic manipulations to validate candidate genes; all distinct advantages over human system genetics. We previously identified increases in OXY state-dependent reward learning and locomotion in female BALB/cJ (J) mice compared to the closely-related BALB/cByJ (ByJ) mice, despite these substrains differing by only ~8,000 genetic variants. These behavioral changes corresponded to increases in the highly potent OXY metabolite oxymorphone (OMOR) within the brain in female Js. A reduced complexity cross between Js and ByJs identified a robust QTL on chromosome 15, accounting for 29% of variance underlying increased whole brain [OMOR] in J females. Further expression QTL analysis using striatal and hippocampal brain tissue identified the transcriptional repressor gene Zhx2 as our top candidate gene. Js contain a mouse endogenous retroviral element (MERV) within Zhx2, decreasing transcription and subsequent protein expression. Zhx2 regulates expression of multiple cytochrome P450 (CYP) enzymes in the liver, the primary site of drug metabolism, and interestingly we observed increases of brain CYP2D11, a mouse ortholog of human CYP2D6 that metabolizes OXY to OMOR, correlated to decreased ZHX2 in female Js. Our findings suggest that decreased ZHX2 expression increases CYP2D expression, consequentially increasing both brain [OMOR] and OXY addiction model behaviors. The primary objective of this proposal is to assess the contribution of Zhx2 to OXY addiction model behaviors through regulation of CYP2D expression and subsequent OMOR production in the brain vs. the liver. In Aim 1, we will identify hepatic transcriptomic and proteomic associations with [OMOR] and OXY addiction model behaviors through RNA-seq and protein mass-spectrometry. In Aim 2, we will delete the Zhx2 MERV through CRISPR/Cas9 gene editing to determine the necessity of this genetic variant in increasing OMOR production and OXY addiction model behaviors. In Aim 3, we will identify the relative role of brain vs. liver Zhx2 in regulating CYP2D expression, OMOR production, and OXY addiction model behaviors through tissue-specific AAV genetic manipulations. Our results will provide critical insight into the genetic, molecular, and tissue-specific mechanisms underlying OXY metabolism and addiction model behaviors that could have implications for personalized medicine in opioid therapeutics and OUD treatments.

项目摘要阿片类药物使用障碍（OUD）仍然是一个主要的公共卫生问题，2020年包含最高数量记录到与Oud相关的死亡。处方阿片类药物有助于大量与Oud相关的死亡，在规定的阿片类药物中，阿片类药物羟考酮（氧）（氧气）是显着的 Ouds的百分比。与Oud相关的特征，包括使用和滥用，行为特征和代谢具有可遗传的组成部分，但基本的OUD特征的遗传变异仍然很大程度上未被发现。啮齿动物基因组具有定量性状基因座（QTL）映射的广泛关联研究（GWAS）可以有效地识别定量特质基因和机制通过强大的样本量，受控环境条件和执行直接遗传操作以验证候选基因的能力；所有不同的优势人类系统遗传学。我们以前确定了氧依赖性的奖励学习和与密切相关的BALB/CBYJ（BYJ）小鼠相比，雌性BALB/CJ（J）小鼠的运动底物仅差异约8,000个遗传变异。这些行为变化对应于增加在女性JS中，大脑中高度有效的氧代谢物矛盾（OMOR）。降低的复杂性交叉在JS和BYJ之间确定了15号染色体上的鲁棒QTL，占基础差异的29％女性的全脑[omor]增加。使用纹状体和海马的进一步表达QTL分析脑组织将转录抑制剂基因ZHX2鉴定为我们的顶级候选基因。 JS包含鼠标 ZHX2内的内源性逆转录病毒元件（MERV），降低转录和随后的蛋白质表达。 ZHX2调节肝脏中多种细胞色素P450（CYP）酶的表达（主要部位）药物代谢，有趣的是，我们观察到脑CYP2D11的增加，人类的小鼠直系同源将Oxy代谢为Omor的CYP2D6与女性JS的ZHX2降低相关。我们的发现表明 ZHX2表达降低会增加CYP2D的表达，从而增加大脑[Omor]和氧成瘾模型行为。该提案的主要目的是评估ZHX2对通过调节CYP2D表达和随后的Omor产生，氧气成瘾模型的行为大脑与肝脏。在AIM 1中，我们将与[Omor]确定肝转录组和蛋白质组学关联通过RNA-seq和蛋白质质谱法的氧气成瘾模型行为。在AIM 2中，我们将删除通过CRISPR/CAS9基因编辑ZHX2 MERV，以确定此遗传变异的必要性增加了Omor的生产和氧气成瘾模型行为。在AIM 3中，我们将确定在调节CYP2D表达，Omor产生和氧成瘾模型行为中，大脑与肝脏ZHX2 通过组织特异性的AAV遗传操作。我们的结果将提供对遗传，分子和组织特异性机制的基础氧代谢和成瘾模型行为，在阿片类药物治疗和OUD治疗中可能对个性化医学有影响。