Cell-specific analysis of sub-kinomes in schizophrenia

项目总结： 这是一个修订的R01应用程序，用于研究锥体神经元信号网络的异常。 精神分裂症。可以轻描淡写地说，分裂谱系障碍的治疗没有 在过去的25年里取得了进展，因为开发了非典型的抗精神病药物。然而，有广泛的 共识是，这些新药不会将药物治疗的疗效扩展到认知 和阴性/缺陷症状，这些症状导致大量患有严重残疾的人 精神分裂症。虽然对精神病或阳性症状有效，但所有抗精神病药物都是 与显著的副作用相关，并有很高的停药率。我们假设这些异常情况 皮质锥体神经元是精神分裂症表型中观察到的许多认知缺陷的基础， 包括工作记忆、执行功能和动机的异常。锥体神经元通常 投射到其他皮质区域(第二层和第三层的浅层锥体细胞)或皮质下(深锥体 第V层和第VI层的细胞)至丘脑、纹状体和其他基底节。发散性异常 精神分裂症患者的浅层和深层锥体神经元可能来自神经发育方面的侮辱 反映这些细胞类型电路的差异，导致基因表达谱改变，神经元 这些神经元的迁移和/或异常连接。我们假设锥体的异常 神经元远远超出了基因表达的简单测量范围，还包括疾病和板层特异性的神经元。 与功能相关的信令网络的变化。为了解决这个问题，我们改编了一部小说《经济学》 用生物信息学方法分析死后脑组织中的丝氨酸/苏氨酸亚组，并鉴定 高产蛋白激酶靶标有待进一步研究。我们建议在此应用程序中重点介绍我们的两个高级- 从这些产生初步研究的假设中产生“命中”：AKT和PKA。具体地说，我们将测试 假设这些激酶在脑内锥体浅层和深层神经元中有不同的调节 精神分裂症。我们将使用一种结合了标准技术的创新方法，包括激光捕获 显微解剖和生化激酶活性分析，以测量锥体神经元特异性激酶 精神分裂症患者的表达和活动。我们将通过测量因子的表达来跟进这些研究 在AKT和PKA下游，包括磷酸蛋白和相关的mRNAs网络。最后，我们 提出了两个离散的电子计算机研究，其中一个专注于开发锥体神经元特有的信号模型 在精神分裂症方面，另一个专注于我们的数据与尖端生物信息学数据库的整合， 确定与标准化病理生理学疾病-药物二联体相关的通路。这些创新之处 研究将确定精神分裂症中锥体神经元特异的信号通路，并提供新的 关于病理生理学的想法和对这种常见疾病的新治疗策略的开发 毁灭性的疾病。

Project Summary: This is a revised R01 application to investigate abnormalities of signaling networks in pyramidal neurons in schizophrenia. It is an understatement to say that the treatment of schizo-spectrum disorders has not progressed in the past 25 years, since the development of atypical antipsychotics. However, there is broad consensus that these newer medications do not extend the efficacy of pharmacological treatments to cognitive and negative/deficit symptoms, which lead to profound disability in a large number of persons afflicted with schizophrenia. While effective for the psychotic, or positive symptoms, all antipsychotic medications are associated with significant side effects, and have high rates of discontinuation. We posit that abnormalities of cortical pyramidal neurons underlie many of the cognitive deficits observed in the schizophrenia phenotype, including abnormalities of working memory, executive function, and motivation. Pyramidal neurons typically project to other cortical regions (superficial pyramidal cells, in layers II and III) or subcortically (deep pyramidal cells, in layers V and VI) to the thalamus, striatum, and other basal ganglia. Divergent abnormalities of superficial and deep pyramidal neurons in schizophrenia may arise from neurodevelopmental insults that reflect differences in the circuitry of these cell types, resulting in altered gene expression profiles, neuronal migration, and/or aberrant connectivity of these neurons. We hypothesize that abnormalities of pyramidal neurons extend well beyond simple measures of gene expression, and include disease- and lamina- specific changes in functionally related signaling networks. To address this problem, we adapted a novel “omics” bioinformatics approach for analysis of serine/threonine sub-kinomes in postmortem brain tissue, and identified high-yield protein kinase targets for further study. We propose to focus in this application on two of our high- yield “hits” from these hypothesis generating preliminary studies: AKT and PKA. Specifically, we will test the hypothesis that these kinases are differentially regulated in superficial and deep pyramidal neurons in schizophrenia. We will use an innovative approach that combines standard techniques, including laser capture microdissection and biochemical kinase activity assays, to measure pyramidal neuron-specific kinase expression and activity in schizophrenia. We will follow up on these studies by measuring expression of factors downstream from AKT and PKA, including phosphoproteins and related networks of mRNAs. Finally, we propose two discrete in silico studies, one focused on developing pyramidal neuron-specific signaling models in schizophrenia, and the other focused on integration of our data with cutting-edge bioinformatics databases, to identify pathways associated with standardized pathophysiological disease-drug dyads. These innovative studies will identify pyramidal neuron-specific signaling pathways disrupted in schizophrenia and provide new ideas regarding the pathophysiology and the development of novel treatment strategies for this often devastating illness.