Controlling FKBP51 for the treatment of PTSD

控制 FKBP51 治疗 PTSD

• 批准号: 10515671
• 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 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10515671
• 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; DNA; Disease; Drops; Drug Targeting; Effectiveness; Ensure; Exposure to; Extinction; FK506 binding protein 5; Feedback; Freedom; Functional disorder; Genes; Glucocorticoids; Goals; Grant; Half-Life; Heat shock proteins; Hormones; Hydrocortisone; Immobilization; 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; 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; preference; prepulse inhibition; promote resilience; 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病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 与压力导致的精神疾病有关51 kDa FK 506结合蛋白FKBP 51， 与90 kDa热休克蛋白（Hsp 90），调节类固醇激素复合物的活性， HPA轴和其他叶栅。FKBP 51-伴侣蛋白复合物减缓HPA的反应 轴循环应激激素。最近，人们发现， 基因中以单核苷酸多态性（SNP）形式出现的遗传变异 编码FKBP 51、FKBP 5，其引起DNA去甲基化和FKBP 5表达增加。 FKBP 5水平已被证明通过类似的机制在应激和衰老期间增加。的 促进去甲基化的SNPs也与压力诱导的糖尿病风险增加有关。 精神病理学，如创伤后应激障碍（PTSD）和重度抑郁症 （MDD）。重要的是，缺乏Fkbp 5的小鼠受到保护，不受与Fkbp 5相关的行为表型的影响。 情绪障碍我们现在已经建立了一种新的转基因小鼠模型， FKBP 5在前脑我们还产生了一种新的细胞系， 标记为FKBP 51，可以很容易地在真实的时间跟踪。通过这些工具，我们将测试 假设降低小鼠中FKBP 51水平的机制将改善对 压力导致的行为缺陷该提案将侧重于1）增加FKBP 51的费率 通过伴侣蛋白调节的蛋白质周转和通过破坏蛋白质表达降低FKBP 5水平 翻译通过使用反义寡核苷酸（ASO）和2）提高我们的理解 FKBP 51如何导致应激诱导的行为缺陷。首先，我们将增加FKBP 51 通过伴侣蛋白调节降解。因为我们知道伴侣蛋白， Hsp 90对蛋白质分类至关重要，我们知道FKBP 51与Hsp 90一起调节类固醇激素 激素复合物，我们假设有一个更大的伴侣蛋白库， 调节FKBP 51的周转。使用表达荧光标记的FKBP 51的细胞，我们将调节 蛋白质伴侣。我们将通过以下方法鉴定新的蛋白伴侣-FKBP 51相互作用： 测量FKBP 51半衰期的变化。我们还将确定FKBP 51的费率和路线 本发明的目的是通过测定FKBP 51的转化率以及评估各种细胞应激物对FKBP 51稳定性的影响来确定FKBP 51的转化率。接下来我们就 测试我们的先导Fkbp 5特异性ASO在野生型小鼠脑中减少Fkbp 5的功效 暴露在压力之下将评价对压力诱导的行为缺陷的保护。最后，在 努力评估FKBP 51对退伍军人报告的最常见PTSD症状的影响， 我们将确定FKBP 51在调节睡眠中断中的作用，睡眠中断是由以下原因引起的： 昼夜节律我们将确定缺乏或过表达FKBP 51的小鼠是否有 改变了昼夜节律的同步性。这些研究的结果可能 揭示FKBP 51靶向治疗可能有利于逆转昼夜节律紊乱， 在创伤后应激障碍以及其他精神疾病和衰老中常见。如果这就是 在这种情况下，那么昼夜节律可以被测量为任何有效性的稳健读数。 FKBP 51靶向疗法的开发。