Controlling FKBP51 for the treatment of PTSD

控制 FKBP51 治疗 PTSD

• 批准号: 9778059
• 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/9778059
• 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; Gap Junctions; 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/antagonist; 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.

美国退伍军人出现精神症状和疾病的风险增加 与平民相比。长期以来，下丘脑-垂体-肾上腺（HPA）轴 与压力引起的精神疾病有关。 51kDa FK506 结合蛋白 FKBP51 一起 与 90kDa 热休克蛋白 (Hsp90) 一起调节类固醇激素复合物的活性 HPA 轴和其他级联。 FKBP51-伴侣复合物减慢 HPA 的反应 轴到循环应激激素。最近发现，天然存在 基因中以单核苷酸多态性 (SNP) 形式出现的遗传变异 编码 FKBP51、FKBP5，导致 DNA 去甲基化和 FKBP5 表达增加。 FKBP5 水平已被证明会通过类似的机制在压力和衰老过程中增加。这 促进去甲基化的 SNP 也与应激诱发的风险增加有关 精神疾病，例如创伤后应激障碍 (PTSD) 和重度抑郁症 （MDD）。重要的是，缺乏 Fkbp5 的小鼠可以免受与 Fkbp5 相关的行为表型的影响。 情绪障碍。我们现在已经生成了一种新型转基因小鼠模型，该模型过度表达 FKBP5 位于前脑。我们还生成了一种过表达荧光的新型细胞系 标记为FKBP51，可以轻松实时追踪。使用这些工具，我们将测试 假设降低小鼠 FKBP51 水平的机制将提高小鼠的适应能力 压力引起的行为缺陷。该提案将重点关注1）提高FKBP51的比率 通过分子伴侣调节蛋白质周转并通过破坏蛋白质降低 FKBP5 水平 通过使用反义寡核苷酸（ASO）进行翻译，2）提高我们对 FKBP51 如何导致压力诱发的行为缺陷。首先我们增加FKBP51 通过伴侣蛋白调节降解。因为我们知道伴侣蛋白，例如 Hsp90 对于蛋白质分类至关重要，我们知道 FKBP51 与 Hsp90 一起调节类固醇 激素复合物，我们假设有一个更大的伴侣蛋白库 调节 FKBP51 营业额。使用表达荧光标记的 FKBP51 的细胞，我们将调节 使用 shRNA 的蛋白质伴侣。我们将通过以下方式鉴定新的蛋白质伴侣-FKBP51 相互作用： 测量 FKBP51 半衰期的变化。我们还将确定FKBP51的费率和路线 周转率以及评估各种细胞应激源对 FKBP51 稳定性的影响。接下来，我们将 测试我们领先的 Fkbp5 特异性 ASO 在减少野生型小鼠大脑中 Fkbp5 方面的功效 承受压力。将评估对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，睡眠中断是由 昼夜节律不同步。我们将确定缺乏或过度表达 FKBP51 的小鼠是否具有 昼夜节律同步性的改变或对压力的反应。这些研究的结果可能 揭示 FKBP51 靶向疗法可能有益于逆转昼夜节律紊乱 常见于创伤后应激障碍以及其他精神疾病和衰老。如果这实际上是 在这种情况下，那么昼夜节律可以被测量为任何有效性的可靠读数 已开发的 FKBP51 靶向疗法。