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: 10675626
• 负责人: TAL KAFRI
• 金额: $ 72.26万
• 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/10675626
• 关键词: ARNTL gene; Acute; Affect; Attention; Behavior; Behavioral; Biodistribution; Biological Process; Blood Pressure; Breeding; Cell Nucleus; Cells; Characteristics; Circadian Dysregulation; Circadian Rhythms; Clock protein; Collection; Core Protein; Crying; Cues; Cultured Cells; Cytoplasm; DNA Repair; Darkness; 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; 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; Repression; 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; circadian; circadian pacemaker; drug addict; gene therapy; genetic association; genomic locus; hormone regulation; immune function; improved; in vivo; knockout gene; light entrainment; 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的每日波动 这些生物过程包括先天性和适应性的 DNA 修复活性。 免疫功能、炎症和代谢途径以自主外周为前提。 振荡核心，存在于大多数器官以及培养的细胞中，是位于体内的中央主时钟。 视交叉上核 (SCN) 通过交感神经同步外周振荡核心 系统和神经内分泌剂如褪黑素（Mel）也被各种夹带。 代谢/激素和身体输入（例如地塞米松、食物摄入量、温度）。 主 SCN 核心通过视网膜下丘脑束接收光输入 所有振荡核心都基于类似的基础。 最近的研究表明，正/负转录/翻译反馈循环对 CR 具有重要影响。 改变小鼠模型中病毒感染的过程并敲除基因。 迄今为止，CR 对疗效的影响高达 10 倍。 我们还没有研究慢病毒载体（LVV）转导的CR是高水平的。 我们提出了三个具体目标来检验这一假设。 将表征正常和中断的 CR 行为对 LVV 肝转导效率的影响。 我们将采用三种表现出不同昼夜节律行为和褪黑激素 (Mel) 产生的小鼠品系，包括 C3H(Mel+)、C57B6 (Mel-) 和 BMAL1（振荡核心蛋白）缺陷型小鼠载体时间的影响。 给药，并且在肝转导中暴露于 Mel 或 Mel 受体拮抗剂将是 将确定改变的 CR 行为模式对载体转导的影响 a) 以下。 C57B6 和 C3H 小鼠中急性阿片类药物戒断方案导致的 CR 破坏，以及 b) 通过基因递送至初始小鼠 在目标 2 中，我们将测试正常日间睡眠时间中断后的 C57B6 和 C3H 小鼠。 假设正常和破坏的 CR 会影响体外人和小鼠细胞的 LVV 转导效率。 具体来说，我们将量化地塞米松夹带对人类和 LVV 转导的影响。 包含正常或突变 BMAL1 基因的小鼠细胞 在目标 3 中，我们将采用 F2 杂交。 CC036 和 CC057 小鼠品系确定之间是否存在遗传关联的效率 LVV 介导的肝脏基因传递和 CR 模式的研究，并将识别导致上述情况的遗传位点 宿主和 LVV 特征。

项目成果

Structural, functional, and immunogenicity implications of F9 gene recoding.

• DOI: 10.1182/bloodadvances.2022007094
• 发表时间: 2022-07-12
• 期刊: BLOOD ADVANCES
• 影响因子: 7.5
• 作者: Katneni, Upendra K.;Alexaki, Aikaterini;Hunt, Ryan C.;Hamasaki-Katagiri, Nobuko;Hettiarachchi, Gaya K.;Kames, Jacob M.;McGill, Joseph R.;Holcomb, David D.;Athey, John C.;Lin, Brian;Parunov, Leonid A.;Kafri, Tal;Lu, Qi;Peters, Robert;V. Ovanesov, Mikhail;Freedberg, Daron I.;Bar, Haim;Komar, Anton A.;Sauna, Zuben E.;Kimchi-Sarfaty, Chava
• 通讯作者: Kimchi-Sarfaty, Chava