Synaptic Protein Networks, Genetic Risk, and Spine Loss in Schizophrenia

精神分裂症的突触蛋白网络、遗传风险和脊柱缺失

• 批准号: 10405455
• 负责人: Matthew L MacDonald
• 金额: $ 48.77万
• 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: 2019-08-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10405455
• 关键词: AMPA Receptors; Acute; Adult; Antipsychotic Agents; Area; Auditory; Auditory Hallucination; Auditory area; Automobile Driving; Autopsy; Biological Assay; Brain; CRISPR/Cas technology; Code; Complex; Computer Analysis; Confocal Microscopy; Dendritic Spines; Dependovirus; Development; Disease model; Drug Targeting; Event; Future; Gene Proteins; Genes; Genetic; Genetic Risk; Genetic study; Genomics; Genotype; Glutamate Receptor; Glutamates; Immune; Impairment; Learning; Link; Maps; Mass Spectrum Analysis; Measures; Mental disorders; Methods; Microscopy; Modeling; Molecular; Monkeys; Mus; Neurobehavioral Manifestations; Neurons; Pathology; Pathway interactions; Phosphorylation; Phosphorylation Site; Post-Translational Protein Processing; Preparation; Process; Protein Analysis; Proteins; Proteomics; Pyramidal Cells; Regulation; Reporting; Role; Schizophrenia; Shotguns; Site; Slice; Synapses; Synaptosomes; TFAP2A gene; Testing; Vertebral column; causal model; cohort; density; disorder risk; drug discovery; experimental study; glutamatergic signaling; gray matter; imaging study; in vivo; in vivo two-photon imaging; innovation; instrumentation; link protein; mature animal; novel; novel therapeutics; paralemmin; postsynaptic; protein expression; protein transport; psychotic symptoms; recruit; risk variant; schizophrenia risk; screening; social cognition; synaptogenesis; trafficking; two photon microscopy

Abstract Schizophrenia (Sz) is a lifelong and devastating psychiatric illness with limited treatment options and no cure. Layer 3 pyramidal cell dendritic spine loss has been repeatedly observed in multiple brain areas in schizophrenia (Sz), including the primary auditory cortex (AI). Spine loss is postulated to underlie primary auditory cortex processing deficits observed in Sz, contributing to impaired social cognition and auditory hallucinations in Sz. We have shown that only smaller spines are lost in Sz Al layer 3. Recent two-photon in vivo imaging studies have shown that new spines are small, essential for synaptogenesis, and required for new learning in adult animals. Dendritic spine formation, stabilization, and plasticity are regulated by synaptic protein network (SynPN) features, such as protein expression, trafficking, and phosphorylation (Phos), and a significant number of Sz risk loci code for synaptic proteins. Targeted and shotgun mass spectrometry (MS) approaches found robust changes in synaptosome and Phos levels of canonical postsynaptic proteins in Sz. These changes were not explained by corresponding changes in homogenate levels of these proteins, suggesting that the brunt of SynPN pathology in Sz is regulated by processes beyond protein expression (e.g. protein trafficking and activity). Nine Phos sites on eight proteins were highly correlated with both synaptosome protein levels and small spine density. All but one of these 8 proteins have well documented roles in vesicular trafficking of postsynaptic glutamate receptors and spine regulation. Postsynaptic glutamate signaling is one of the most significantly implicated pathways in genetic studies of Sz. Thus, we hypothesize that: Aberrant trafficking and Phos of postsynaptic proteins is linked to Sz genetic risk and drives small spine loss in Sz Al. We will test this hypothesis in an unprecedented set of parallel genomic, proteomic, and microscopy experiments in 100 Sz and 100 matched control subjects with cutting edge computational analyses to identify protein and Phos linked to Sz genetics and generate causal models of disease (Aim 1). We will then utilize innovative molecular and two-photon microscopy approaches to test the effects of candidate Phos, from our preliminary studies and high priority Aim 1 findings, on spine density, size, formation, and stability in the Al of adult mice (Aim 2). These studies will identify proximal molecular events, potentially associated with Sz risk genetics, that impair small spines in Sz Al, as well as the stage of spine formation/stabilization that is impaired. Such events can be further investigated in vivo via CRISPR/Cas9 in future studies and have the potential to serve as targets for the development of novel therapeutics.

抽象的 精神分裂症（SZ）是一种终生而毁灭性的精神病，治疗方案有限，无法治愈。 在精神分裂症的多个大脑区域中，已经反复观察到第3层锥体细胞树突状脊柱丧失 （SZ），包括主要的听觉皮层（AI）。脊柱损失被认为是主要听觉皮层的基础 在SZ中观察到的处理缺陷，导致SZ社会认知和听觉幻觉受损。 我们已经表明，仅在SZ Al第3层中丢失了较小的棘突。最近的两光体体内成像研究 已经表明新刺很小，对于突触发生必不可少，成人新学习必需 动物。树突状脊柱形成，稳定和可塑性受突触蛋白网络（SYNPN）调节 特征，例如蛋白质表达，运输和磷酸化（PHO），以及大量的SZ风险 突触蛋白的基因座代码。 靶向和shot弹枪质谱法（MS）方法发现了突触体和PHO的强大变化 SZ中规范突触后蛋白的水平。这些变化没有通过相应的更改来解释 在这些蛋白质的匀浆水平中，表明SZ中的Synpn病理的首当其 超出蛋白质表达的过程（例如蛋白质运输和活性）。八个蛋白质上的九个PHO位点 与突触体蛋白水平和小脊柱密度高度相关。除了这8个 蛋白质在突触后谷氨酸受体和脊柱的囊泡运输中具有充分的作用 规定。突触后谷氨酸信号传导是遗传中最有意义的途径之一 SZ的研究。因此，我们假设：与突触后蛋白的异常运输和PHO相连 SZ遗传风险并驱动SZ AL中的小脊柱损失。 我们将在前所未有的平行基因组，蛋白质组学和显微镜中检验该假设 在100 sz和100个匹配的对照对象中进行的实验，并具有尖端计算分析以识别 蛋白质和PHO与SZ遗传学有关，并产生疾病的因果模型（AIM 1）。然后我们将使用 从我们的 初步研究和高优先级目标1发现，关于脊柱密度，大小，形成和稳定性的发现 成年小鼠（目标2）。 这些研究将确定近端分子事件，可能与SZ风险遗传学有关，这会损害 SZ Al中的小刺以及受损的脊柱形成/稳定阶段。这样的事件可以是 在未来的研究中通过CRISPR/CAS9进一步研究了体内，并有可能作为目标的目标 新型治疗学的发展。