The STAT3 response of excitatory neurons to epileptogenic brain injury

兴奋性神经元对致癫痫性脑损伤的 STAT3 反应

• 批准号: 10119388
• 负责人: Amy R. Brooks-Kayal
• 金额: $ 57.86万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-15 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10119388
• 关键词: Affect; Anabolism; Animals; Antibodies; Apoptosis; Astrocytes; Behavioral; Brain; Brain Injuries; Ca(2+)-Calmodulin Dependent Protein Kinase; Cell Nucleus; Cells; Ceramides; ChIP-seq; Complex; Contralateral; Data Set; Development; Disease; Electroencephalography; Epilepsy; Epileptogenesis; Frequencies; Future; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genome; Hippocampus (Brain); Hour; Hyperactive behavior; Immunity; Individual; Inflammation; Injections; Interneurons; Intervention; Intractable Epilepsy; Ion Channel Gating; Ipsilateral; Janus kinase; Laboratories; Lead; Ligands; Long-Term Depression; Measures; Membrane; Memory; Memory impairment; Metabolism; Microglia; Modeling; Molecular; Molecular Profiling; Monitor; Mus; Neurons; Pathway interactions; Patients; Pharmacology; Pilocarpine; Population; Predisposition; Process; Prosencephalon; Reporting; Role; STAT protein; STAT3 gene; Seizures; Severities; Signal Transduction; Site; Sphingolipids; Status Epilepticus; Structure; Symptoms; Synaptic plasticity; Tamoxifen; Technology; Temporal Lobe Epilepsy; Testing; Time; Transcriptional Regulation; Transgenic Organisms; Up-Regulation; Video Recording; brain tissue; calmodulin-dependent protein kinase II; cell type; conditioned fear; early detection biomarkers; excitatory neuron; granule cell; hippocampal pyramidal neuron; in vivo; inhibitor/antagonist; kainate; knock-down; metabolome; metabolomics; mouse model; neural circuit; neuroprotection; promoter; receptor binding; relating to nervous system; response; trafficking; transcriptome; transcriptome sequencing; transcriptomics; voltage

ABSTRACT Temporal lobe epilepsy (TLE) develops after a period of ongoing molecular cascades and neural circuit remodeling in the hippocampus resulting in increased susceptibility to spontaneous seizures. Targeting these cascades in TLE patients could reverse their symptoms and have the potential to provide a viable disease- modifying treatment, especially for the large portion of over 30% of TLE patients who do not respond to any available treatments. In recent years, the Janus Kinase/Signal Transducer and Activator of Transcription (JAK/STAT) pathway has been implicated in temporal lobe epilepsy (TLE). The JAK/STAT pathway is known to be involved in inflammation and immunity, and only more recently has been shown to be associated with neuronal functions such as synaptic plasticity. Our laboratories previously showed that a JAK inhibitor, WP1066, could greatly reduce the number of spontaneous seizures that animals went on to develop over time in the pilocarpine model of status epilepticus (SE). We have continued to investigate the mechanism of JAK/STAT- induced epileptogenic responses through the use of a new transgenic line we developed where STAT3 knockdown (KD) can be controlled by tamoxifen-induced CRE expression specifically in forebrain excitatory neurons via the Calcium/Calmodulin Dependent Protein Kinase II alpha (CamK2a) promoter. We now report that this knockdown of STAT3 (nSTAT3KD) markedly reduces spontaneous seizure frequency in the intrahippocampal kainate model (IHKA) and “rescues” mice from KA-induced memory deficits as measured by Contextual Fear Conditioning. Recently, using deep RNA-sequencing we also discovered transcriptomic signatures 24 hours after SE that occur in response to IHKA injections (ipsilateral and contralateral to the injection site) and are reversed by nSTAT3KD, especially for those genes important in sphingolipid metabolism: a regulator of neuronal structure, and the trafficking, stability, and function of multiple membrane bound receptors, including ligand- and voltage-gated ion channels. These findings, taken together with our preliminary IHKA metabolome, brings us to propose the following unique hypothesis that there is a JAKx/STAT3 pathway in excitatory forebrain neurons that becomes activated in response to prolonged seizures and that identifying the cells most susceptible to STAT3 signaling during the epileptogenic process will provide a window on basic circuitry that underlies memory formation, and most importantly, the brain's susceptibility to epilepsy development. To test this hypothesis, we have three Aims using state of the art molecular technologies (metabolomic profiling, single nuclei RNA sequencing, and chromatin immunoprecipitation sequencing) to interrogate the molecular signature of the hippocampus (24 h, 2 wk, and 4 wk after IHKA SE) . The emerging transcriptome for STAT3 in the context of epilepsy suggests that it may be useful for identifying potential epileptogenic gene networks that were previously unknown, selecting early-detection biomarkers that inform seizure susceptibility, as well as choosing new targets for the future treatment of intractable epilepsies.

抽象的 颞叶癫痫（TLE）在持续的分子级联反应和神经回路后发展 海马中的重塑，导致对赞助癫痫发作的敏感性增加。针对这些 TLE患者的级联反应可能扭转其症状，并有可能提供可行的疾病 - 修改治疗 可用的治疗。近年来，Janus激酶/信号换能器和转录激活剂 （JAK/STAT）途径已在临时叶癫痫（TLE）中隐含。已知JAK/STAT途径 参与炎症和免疫力，最近才显示与 神经元功能，例如突触可塑性。我们的实验室以前表明JAK抑制剂WP1066， 可以大大减少动物随着时间的流逝而在 状态癫痫症（SE）的毛果石模型。我们继续研究jak/stat-的机制 通过使用新的转基因线，我们开发了STAT3的诱导癫痫反应 敲低（KD）可以通过他莫昔芬诱导的Cre表达来控制前脑兴奋 通过钙/钙调蛋白依赖性蛋白激酶IIα（CAMK2A）启动子神经元的神经元。我们现在报告 STAT3（NSTAT3KD）的这种敲低可显着降低 Hapocampal Kainate模型（IHKA）和从KA诱导的记忆中的“救援”小鼠定义了 上下文恐惧条件。最近，使用深RNA - 我们还发现了转录组 SE后24小时对IHKA注射（同侧和对侧注射）的签名 位置），并由NSTAT3KD逆转，特别是对于那些在鞘脂代谢中重要的基因： 神经元结构的调节剂，以及多个膜结合接收器的运输，稳定和功能， 包括配体和电压门控离子通道。这些发现，与我们的初步IHKA一起 代谢组使我们提出以下独特的假设，即在 兴奋性的前脑神经元会因长期癫痫发作而被激活，并鉴定 在癫痫发作过程中最容易受到STAT3信号传导的细胞将为基本提供一个窗口 是记忆形成的基础的电路，最重要的是大脑对癫痫的敏感性 发展。为了检验这一假设，我们使用最先进的分子技术有三个目标 （代谢组分析，单核RNA测序和染色质免疫沉淀测序）至 询问海马的分子特征（Ihka SE之后的24h，2WK和4WK）。新兴 STAT3在癫痫的背景下的转录组表明，它可能有助于识别潜力 以前未知的癫痫发作基因网络，选择早期检测生物标志物来告知 癫痫发作的敏感性，以及为未来治疗顽固发作的新目标选择新目标。