Pyramidal neuron hypoactivity in schizophrenia cortex

精神分裂症皮质锥体神经元活性低下

• 批准号: 10452726
• 负责人: Kirsten Schoonover
• 金额: $ 8.78万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10452726
• 关键词: ATP phosphohydrolase; Address; Affect; Alleles; Animals; Antibodies; Antipsychotic Agents; Autopsy; Behavior; Behavioral; Binding Proteins; Biological Assay; Biological Markers; Blood; Blood - brain barrier anatomy; Brain; CREB1 gene; Candidate Disease Gene; Carrier Proteins; Cell Culture Techniques; Cognitive deficits; Collection; Conflict (Psychology); Copper; Data; Data Collection; Delusions; Demyelinations; Disease; Dopamine; Dopamine-beta-monooxygenase; Drug Targeting; Educational process of instructing; Electron Microscopy; Epigenetic Process; Exhibits; Family; Fiber; Foundations; Future; Genes; Genetic; Genotype; Goals; Hallucinations; Hippocampus (Brain); Homeostasis; Hypermethylation; Immunohistochemistry; Impaired cognition; Impairment; Individual; Knock-out; Knockout Mice; Knowledge; Learning; Learning Skill; Link; MAP Kinase Gene; Mediating; Mediation; Mental disorders; Mentors; Messenger RNA; Metabolism; Methylation; Midbrain structure; Mitochondria; Molecular; Mus; Neuraxis; Pathology; Pathway interactions; Patients; Peripheral; Persons; Population; Protein Deficiency; Protein Isoforms; Proteins; Regulation; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Schizophrenia; Starvation; Substantia nigra structure; Symptoms; System; Techniques; Testing; Tissues; Variant; Western Blotting; Wild Type Mouse; Writing; behavioral impairment; brain volume; cognitive function; drug development; dystrobrevin; emerging adult; experimental study; gene therapy; genetic variant; high risk; hippocampal pyramidal neuron; hypocupremia; interest; learning progression; meetings; monoamine; mouse model; multidisciplinary; myelination; new therapeutic target; novel marker; preadolescence; promoter; protein expression; quetiapine; risk variant; skills; stem; white matter

Schizopphrenia (SZ) affects 1% of the world population. Allelic variants of the dystrobrevin binding protein 1 (DTNBP1) gene, now a top SZ candidate gene, are associated with fiber tract impairments in healthy people and reduced brain volume in preteens- both SZ symptoms. However, the downstream effects of decreased dysbindin are poorly understood. Dysfunctional dysbindin decreases the copper-transporting P-type ATPase (ATP7A) and the copper transporter CTR1. ATP7A and CTR1 facilitate copper transport between the blood and the brain. The role of copper in SZ has been controversial, with many studies finding increased blood copper in patients. However, these studies have not considered copper may build up in the blood because it cannot get into the brain. Copper-decreasing experimental manipulations produce demyelination, dopamine increases, and SZ-like behavioral impairments. Taken together, dysbindin/copper alterations can result in SZ- like pathology such as impaired white matter integrity, excess dopamine, and altered mitochondrial activity, and yet this combined pathway remains largely unstudied. Overall, I hypothesize copper transport alterations contribute to SZ pathology, potentially through decreased dysbindin expression. I will test this in postmortem SZ brain, first-episode antipsychotic-naïve SZ patients, and a dysbindin knockout mouse model. Specific Aim 1: I have learned western blotting, and am now learning immunohistochemical analysis. Specific Aim 2A. We will test the copper state of postmortem substantia nigra and hippocampus, and the relationship of this state to copper transporters ATP7A/CTR1 in both treated and untreated SZ. Furthermore, we will assess the mechanism of action of antipsychotics in relation to these variables. Specific Aim 2B. To discover potential schizophrenia biomarkers, we will assess peripheral copper state and its relationship to copper transporters and methylation status of DTNBP1, SLC31A1(CTR1), and ATP7A before and after 6 weeks of antipsychotic treatment in first-episode, antipsychotic-naïve SZ patients. Specific Aim 2C. In dysbindin knockout, heterozygous, and wild-type mice, we will test peripheral, hippocampal, and nigral copper state, and the relation to copper transporters ATP7A/CTR1, cognitive deficits, and the mechanism of antipsychotic mediated changes before and after 28 days quetiapine treatment. Specific Aim 3: These studies will provide the first evidence of SZ brain copper state, and resolve the juxtaposition between excess copper in SZ blood and the SZ-like symptoms induced by copper deficit. Furthermore, these studies will provide new data about risk gene dysbindin and its effects on copper transporters (a previously unstudied pathway in SZ) and a potential mechanism of antipsychotic rescue of copper starvation deficits, which could yield novel targets for drug development. I will learn quantitative RT- PCR, methylation assay, immunohistochemistry, electron microscopy and tissue sectioning, brain collection, scientific writing, lab and animal management, and all that these skills entail, as well as proficient teaching.

精神分裂症 (SZ) 影响着世界人口的 1%。抗坏血酸结合蛋白的等位基因变体 1 (DTNBP1) 基因目前是 SZ 的顶级候选基因，与健康人的纤维束损伤有关 青春期前的脑容量减少——这都是精神分裂症的症状。然而，下游影响减弱 人们对dysbindin知之甚少。功能失调的 Dysbindin 会降低铜转运 P 型 ATP 酶 (ATP7A) 和铜转运蛋白 CTR1。 ATP7A 和 CTR1 促进血液之间的铜转运 和大脑。铜在 SZ 中的作用一直存在争议，许多研究发现增加血液 患者体内的铜。然而，这些研究并没有考虑到铜可能会在血液中积聚，因为它 无法进入大脑。减少铜的实验操作会产生脱髓鞘、多巴胺 增加，以及类似 SZ 的行为障碍。综上所述，dysbindin/铜的改变可导致 SZ- 例如白质完整性受损、多巴胺过多和线粒体活性改变等病理学， 然而，这种组合途径在很大程度上仍未得到研究。总的来说，我假设铜的运输发生了变化 可能通过减少 Dysbindin 表达来促进 SZ 病理。我将在事后测试这一点 SZ 大脑、首次接受抗精神病药物治疗的 SZ 患者以及 Dysbindin 敲除小鼠模型。 具体目标1：我已经学习了蛋白质印迹，现在正在学习免疫组化分析。 具体目标 2A。我们将测试死后黑质和海马体的铜状态，以及 在处理和未处理的 SZ 中，这种状态与铜转运蛋白 ATP7A/CTR1 的关系。此外， 我们将评估与这些变量相关的抗精神病药物的作用机制。 具体目标 2B。为了发现潜在的精神分裂症生物标志物，我们将评估外周铜状态 及其与铜转运蛋白和 DTNBP1、SLC31A1(CTR1) 和 ATP7A 甲基化状态的关系 首发、未接受抗精神病药物治疗的 SZ 患者接受抗精神病药物治疗 6 周之前和之后。 具体目标 2C。在 Dysbindin 敲除、杂合和野生型小鼠中，我们将测试外周、 海马和黑质铜状态，以及与铜转运蛋白 ATP7A/CTR1、认知缺陷、 以及 28 天喹硫平治疗前后抗精神病药物介导的变化机制。 具体目标 3：这些研究将提供 SZ 脑铜状态的第一个证据，并解决 SZ 血液中过量的铜与铜缺乏引起的 SZ 样症状并存。 此外，这些研究将提供有关风险基因失调及其对铜的影响的新数据 转运蛋白（深圳以前未研究过的途径）和抗精神病药物救援的潜在机制 铜匮乏，这可能会产生药物开发的新目标。我将学习定量RT- PCR、甲基化测定、免疫组织化学、电子显微镜和组织切片、脑采集、 科学写作、实验室和动物管理，以及所有这些技能所需要的，以及熟练的教学。