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: 10618896
• 负责人: Matthew L MacDonald
• 金额: $ 48.02万
• 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 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10618896
• 关键词: AMPA Receptors; Acute; Adult; Antipsychotic Agents; Area; 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; Exclusion; 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; Postsynaptic Membrane; Preparation; Process; Proteins; Proteomics; Pyramidal Cells; Regulation; Reporting; Role; Schizophrenia; Shotguns; Site; Slice; Sorting; 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; mimetics; 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) 是一种终生且具有毁灭性的精神疾病，治疗选择有限且无法治愈。 在精神分裂症患者的多个脑区反复观察到第 3 层锥体细胞树突棘缺失 （Sz），包括初级听觉皮层（AI）。脊柱损失被认为是初级听觉皮层的基础 在 Sz 中观察到的处理缺陷，导致 Sz 中的社会认知受损和幻听。 我们已经证明，Sz Al 第 3 层中仅丢失了较小的脊柱。最近的双光子体内成像研究 研究表明，新的棘很小，对于突触发生至关重要，并且是成人新学习所必需的 动物。树突棘的形成、稳定性和可塑性由突触蛋白网络 (SynPN) 调节 特征，例如蛋白质表达、运输和磷酸化 (Phos)，以及大量 Sz 风险 突触蛋白的基因座代码。 靶向和鸟枪质谱 (MS) 方法发现突触体和 Phos 发生了剧烈变化 Sz 中典型突触后蛋白的水平。这些变化没有通过相应的变化来解释 这些蛋白质的匀浆水平，表明 Sz 中 SynPN 病理学的主要影响是由 蛋白质表达之外的过程（例如蛋白质运输和活性）。八种蛋白质上的九个磷酸位点 与突触体蛋白水平和小棘密度高度相关。除了这 8 个中的一个之外，所有其他 蛋白质在突触后谷氨酸受体和脊柱的囊泡运输中的作用已得到充分证明 规定。突触后谷氨酸信号传导是遗传中最重要的相关途径之一 Sz 的研究。因此，我们假设： 突触后蛋白的异常运输和 Phos 相关 Sz 遗传风险并导致 Sz Al 的脊柱出现轻微损失。 我们将在前所未有的平行基因组学、蛋白质组学和显微镜检查中检验这一假设 在 100 Sz 和 100 名匹配对照受试者中进行的实验，通过尖端计算分析来识别 与 Sz 遗传学相关的蛋白质和 Phos 并生成疾病的因果模型（目标 1）。然后我们将利用 创新的分子和双光子显微镜方法来测试候选磷酸盐的效果，来自我们 关于 Al 中脊柱密度、大小、形成和稳定性的初步研究和高度优先的目标 1 结果 成年小鼠（目标 2）。 这些研究将确定可能与 Sz 风险遗传学相关的近端分子事件，这些事件会损害 Sz Al 中的小刺，以及受损的刺形成/稳定阶段。此类事件可以是 在未来的研究中通过 CRISPR/Cas9 进一步进行体内研究，并有可能作为 新疗法的开发。