The circadian rhythm as a lentiviral vector restriction factor

昼夜节律作为慢病毒载体的限制因素

• 批准号: 10475046
• 负责人: TAL KAFRI
• 金额: $ 73.97万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10475046
• 关键词: ARNTL gene; Acute; Affect; Attention; Behavior; Behavioral; Biodistribution; Biological Process; Blood Pressure; Breeding; Cell Nucleus; Cells; Characteristics; Circadian Rhythms; Clock protein; Collection; Core Protein; Cues; Cultured Cells; Cytoplasm; DNA Repair; Data; Dexamethasone; Drug usage; Eating; Epidemic; Etiology; Eukaryotic Cell; Exhibits; Exposure to; Feedback; Gene Delivery; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Genetic Variation; Goals; Hepatic; High Prevalence; Hormonal; Host resistance; Human; Hypothalamic structure; In Vitro; Inbred C3H Mice; Infection; Inflammation; Invaded; Knock-out; Laboratories; Lentivirus Vector; Light; Malignant Neoplasms; Mediating; Melatonin; Melatonin Receptors; Metabolic; Metabolic Diseases; Metabolic Pathway; Molecular; Motor Activity; Mouse Strains; Mus; Neurodegenerative Disorders; Neurons; Neurosecretory Systems; Organ; Orphan; Output; Pathology; Pathway interactions; Pattern; Peripheral; Phenotype; Phosphorylation; Physiological; Physiological Processes; Production; Proteins; Protocols documentation; Receptor Gene; Retina; Retinoids; Signal Transduction; Simplexvirus; Sleep; Societies; Substance abuse problem; Sympathetic Nervous System; Temperature; Testing; Time; Transcriptional Activation; Translating; Translations; Untranslated RNA; Viral; Viral Vector; Virus Diseases; Work; addiction; antagonist; arm; base; circadian; circadian pacemaker; drug addict; gene therapy; genetic association; genomic locus; hormone regulation; immune function; improved; in vivo; mouse model; mutant; opioid withdrawal; paracrine; pathogen; permissiveness; sleep pattern; suprachiasmatic nucleus; transduction efficiency; vector

Abstract: Circadian rhythm (CR) defines the daily oscillation in gene expression that controls major physiologic pathways in eukaryotic cells in vivo and in vitro. These biological processes include DNA repair activity, innate and adaptive immune functions, inflammation, and metabolic pathways. CRs are premised on autonomous peripheral oscillatory cores, which exist in most organs as well as in cultured cells. In vivo, a central master clock located in the suprachiasmatic nucleus (SCN) synchronizes the peripheral oscillatory cores via the sympathetic nervous system and neuroendocrine agents such as melatonin (Mel). Peripheral organs are also entrained by various metabolic/hormonal and physical inputs (e.g. Dexamethasone, food intake, temperature). However, only the master SCN core receives light inputs via the retinal hypothalamic tract. All oscillatory cores are based on similar positive/negative transcriptional/translational feedback loops. Recent studies demonstrated major CR effects on the course of viral infections in murine models. Altering the time of viral application and knocking out genes encoding oscillatory core proteins enhanced viral infection by up to 10-fold. To date, CR effects on the efficacy of Lentiviral vector (LVV) transduction have not been studied. We hypothesize that the CR is a high-level restriction factor to viral-vector transduction. We propose three specific aims to test this hypothesis. In aim 1, we will characterize the effects of normal and disrupted CR behavior on hepatic transduction efficiency by LVVs. We will employ three mouse strains showing distinct circadian behavior and melatonin (Mel) production, including C3H(Mel+), C57B6 (Mel-), and BMAL1 (oscillatory core protein)-deficient mice. The effects of time of vector administration, and exposure to either Mel or the Mel- receptor antagonist on hepatic transduction will be determined. The effects of altered CR behavior patterns on vector transduction will be determined a) following CR disruption by an acute opioid withdrawal protocol in C57B6 and C3H mice, and b) by gene delivery to naïve C57B6 and C3H mice following disruption of the normal day-sleep time period. In aim 2, We will test the hypothesis that normal and disrupted CRs affect LVV transduction efficiency in human and mouse cells in vitro. Specifically, we will quantify the effects of dexamethasone entrainment on LVV transduction of human and mouse cells comprising either normal or mutant BMAL1 gene. In aim 3, We will employ an F2 cross between the CC036 and CC057 mouse strains to determine the existence of a genetic association between the efficiency of LVV mediated hepatic gene delivery and CR patterns and will identify genetic loci contributing to the above host and LVV characteristics.

抽象的： 昼夜节律（CR）定义了控制主要生理途径的基因表达中的每日振荡 在体内和体外的真核细胞中。这些生物学过程包括DNA修复活动，先天和自适应 免疫功能，炎症和代谢途径。 CRS以自主外围为前提 振荡核心，在大多数器官以及培养的细胞中存在。 in Vivo，一个中央大师时钟 在超节核（SCN）中，通过交感神经系统同步外周振荡核心 系统和神经内分泌剂，例如褪黑激素（MEL）。外围器官也被各种 代谢/荷尔蒙和物理输入（例如地塞米松，食物摄入，温度）。但是，只有 主SCN核心通过视网膜下丘脑接收光输入。所有振荡核心均基于相似的 正/负转录/翻译反馈回路。最近的研究表明了对CR的主要影响 鼠模型中的病毒感染过程。改变病毒应用的时间并淘汰基因 编码振荡性核心蛋白可增强病毒感染多达10倍。迄今为止，CR对有效性的影响 慢病毒载体（LVV）翻译的翻译尚未研究。我们假设CR是高级 病毒载体转移的限制因子。我们提出了三个特定的目的来检验这一假设。在AIM 1中，我们 将表征正常和破坏CR行为对LVVS肝翻译效率的影响。 我们将使用三种鼠标菌株显示出不同的昼夜节律行为和褪黑激素（MEL）的产生，包括 C3H（MEL+），C57B6（MEL-）和BMAL1（振荡性核心蛋白）缺陷小鼠。向量时间的影响 给药，并在肝翻译上接触MEL或MEL受体拮抗剂 决定。 CR行为模式改变对向量翻译的影响将确定a） C57B6和C3H小鼠中急性阿片类药物戒断方案的CR破坏，b）基因递送至幼稚 正常日睡眠时间中断后，C57B6和C3H小鼠。在AIM 2中，我们将测试 假设正常和破坏的CR会影响人体和小鼠细胞的LVV转导效率。 具体而言，我们将量化地塞米松融化对人类LVV转导的影响 完成正常或突变BMAL1基因的小鼠细胞。在AIM 3中，我们将使用F2交叉 CC036和CC057小鼠菌株，以确定效率之间存在遗传关联 LVV介导的肝基因递送和CR模式的介导，将确定对上述贡献的遗传基因座 宿主和LVV特征。