Controlling FKBP51 for the treatment of PTSD

控制 FKBP51 治疗 PTSD

• 批准号: 10421246
• 负责人: Laura J Blair
• 金额: --
• 依托单位: JAMES A. HALEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-10-01 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10421246
• 关键词: Aging; Animal Model; Antisense Oligonucleotides; Antisense Technology; Behavior; Behavior assessment; Behavioral; Biological Assay; Biology; Brain; Cell Line; Cells; Circadian Dysregulation; Circadian Rhythm Sleep Disorders; Circadian Rhythms; Circadian desynchrony; Complex; Conflict (Psychology); DNA; Disease; Drops; Drug Targeting; Effectiveness; Ensure; Exposure to; Extinction (Psychology); FK506 binding protein 5; Feedback; Freedom; Functional disorder; Genes; Glucocorticoids; Goals; Grant; Half-Life; Heat shock proteins; Hormones; Hydrocortisone; In Vitro; Incidence; Knowledge; Label; Lead; Libraries; Link; Major Depressive Disorder; Measures; Mediating; Mental Depression; Mental disorders; Military Personnel; Molecular Chaperones; Mood Disorders; Mus; Neurons; Oligonucleotides; Periodicity; Population; Post-Traumatic Stress Disorders; Predisposition; Prosencephalon; Proteins; Psychopathology; Regulation; Reporting; Resistance; Response Elements; Risk; Role; Route; Serum; Shock; Single Nucleotide Polymorphism; Sleep disturbances; Soldier; Stress; Sucrose; Symptoms; System; Tacrolimus Binding Proteins; Tail Suspension; Testing; Time; Transgenic Mice; Translations; Triage; Veterans; Wild Type Mouse; Work; base; behavioral phenotyping; biological adaptation to stress; chaperone machinery; circadian; common symptom; comorbidity; conditioned fear; demethylation; design; genetic variant; hypothalamic-pituitary-adrenal axis; improved; in vivo; inhibitor; knock-down; mouse model; multicatalytic endopeptidase complex; novel; novel strategies; operation; overexpression; photo switch; preference; prepulse inhibition; protein degradation; psychiatric symptom; psychologic; resilience; response; sleep regulation; small hairpin RNA; steroid hormone; stress resilience; stressor; targeted treatment; therapy development; tool; wound

U.S. Veterans are at increased risk for developing psychiatric symptoms and disorders compared to the civilian population. The hypothalamic-pituitary-adrenal (HPA) axis has long been linked to stress-induced psychiatric disorders. The 51kDa FK506-binding protein, FKBP51, together with the 90kDa heat shock protein (Hsp90), regulates the activity of steroid hormone complexes in the HPA axis and other cascades. The FKBP51-chaperone complex slows the response of the HPA axis to circulating stress hormones. Recently, it has been discovered that there are naturally occurring genetic variants in the form of single nucleotide polymorphisms (SNPs) in the gene encoding FKBP51, FKBP5, that cause DNA demethylation and increased expression of FKBP5. FKBP5 levels have been shown to increase during stress and aging by a similar mechanism. The SNPs that promote demethylation are also associated with increased risk for stress-induced psychopathologies such as post-traumatic stress disorder (PTSD) and major depressive disorder (MDD). Importantly, mice lacking Fkbp5 are protected from behavioral phenotypes associated with mood disorders. We have now generated a novel transgenic mouse model that overexpresses FKBP5 in the forebrain. We have also generated a novel cell line which overexpresses fluorescently labelled FKBP51, which can be easily tracked in real time. With these tools, we will test the hypothesis that mechanisms which decrease the levels of FKBP51 in mice will improve resiliency to stress-induced behavioral deficits. This proposal will focus on 1) increasing the rate of FKBP51 protein turnover through chaperone regulation and decreasing FKBP5 levels by disrupting protein translation through use of antisense oligonucleotides (ASOs) and 2) improving our understanding of how FKBP51 contributes to stress-induced behavioral deficits. First, we will increase FKBP51 degradation through chaperone protein modulation. Since we know that chaperone proteins, like Hsp90, are vital for protein triage and we know that FKBP51 works with Hsp90 to regulate steroid hormone complexes, we hypothesize that there is a larger chaperone protein repertoire which regulates FKBP51 turnover. Using cells expressing fluorescently tagged FKBP51, we will modulate protein chaperones using shRNA. We will identify novel protein chaperone-FKBP51 interactions by measuring changes in FKBP51 half-life. We will also determine the rate and route of FKBP51 turnover as well as assess the impact of various cellular stressors on FKBP51 stability. Next, we will test our lead Fkbp5-specific ASOs for their efficacy in reducing Fkbp5 in the brain of wild-type mice exposed to stress. Protection from stress-induced behavioral deficits will be evaluated. Lastly, in an effort to evaluate the impact of FKBP51 on the most common PTSD symptom reported by Veterans, we will determine the role of FKBP51 in the regulation of sleep disruption, which is caused by circadian desynchrony. We will determine whether mice lacking or overexpressing FKBP51 have altered circadian synchrony basally or in response to stress. The results from these studies may reveal that FKBP51-targeted therapies could be beneficial for reversing the circadian disruption that is commonly found in PTSD as well as in other psychiatric disorders and aging. If this is in fact the case, then circadian rhythmicity could be measured as a robust readout for the effectiveness of any FKBP51-targeted therapies that are developed.

美国退伍军人患精神症状和疾病的风险增加 与平民相比。下丘脑 - 垂体 - 肾上腺（HPA）轴长期以来一直是 与压力引起的精神疾病有关。 51KDA FK506结合蛋白FKBP51一起 使用90KDA热休克蛋白（HSP90）调节类固醇激素复合物在 HPA轴和其他级联。 FKBP51-CHAPERONE复合物减慢了HPA的响应 轴向循环应力激素。最近，已经发现自然存在 在基因中以单核苷酸多态性（SNP）的形式出现的遗传变异 编码FKBP51，FKBP5，导致DNA脱甲基化并增加了FKBP5的表达。 通过类似机制，已显示在压力和衰老期间FKBP5水平增加。这 促进脱甲基化的SNP也与增加压力引起的风险增加有关 心理病理学，例如创伤后应激障碍（PTSD）和主要抑郁症 （MDD）。重要的是，缺乏FKBP5的小鼠免受与行为表型的保护 情绪障碍。现在，我们已经生成了一种过表达的新型转基因小鼠模型 FKBP5在前脑。我们还产生了一种新型的细胞系，该细胞系过表达荧光 标记为FKBP51，可以实时轻松跟踪。使用这些工具，我们将测试 假设降低小鼠FKBP51水平的机制将提高弹性 压力引起的行为缺陷。该提案将重点是1）提高FKBP51的速度 蛋白质通过伴侣调节的蛋白质更新，并通过破坏蛋白质而降低FKBP5水平 通过使用反义寡核苷酸（ASO）和2）转化 FKBP51如何导致压力引起的行为缺陷。首先，我们将增加FKBP51 通过伴侣蛋白调节的降解。由于我们知道伴侣蛋白，就像 HSP90对于蛋白质分类至关重要，我们知道FKBP51可与HSP90一起调节类固醇 激素复合物，我们假设有一个较大的伴侣蛋白库 调节FKBP51营业额。使用表达荧光标记的FKBP51的单元，我们将调制 使用shRNA的蛋白质伴侣。我们将通过 测量FKBP51半衰期的变化。我们还将确定FKBP51的速率和途径 营业额以及评估各种细胞应激源对FKBP51稳定性的影响。接下来，我们会的 测试我们的铅FKBP5特异性ASO，以减少野生型小鼠大脑中的FKBP5的功效 暴露于压力。将评估免受压力引起的行为缺陷的保护。最后，在一个 努力评估FKBP51对退伍军人报告的最常见PTSD症状的影响， 我们将确定FKBP51在调节睡眠中断中的作用，这是由 昼夜节律。我们将确定缺乏或过表达FKBP51的小鼠具有 基本或响应应力改变了昼夜节律同步。这些研究的结果可能 揭示以FKBP51为目标的疗法可能有益于扭转昼夜节律的破坏 通常在PTSD以及其他精神疾病和衰老中发现。如果这实际上是 案例，然后可以测量昼夜节律的节奏，以衡量任何的读数 开发的FKBP51靶向疗法。