Single Nucleus Transcriptional Profiling of Intractable Focal Epilepsy

难治性局灶性癫痫的单核转录谱

• 批准号: 10373149
• 负责人: Peter Canoll
• 金额: $ 23.53万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373149
• 关键词: Area; Astrocytes; Biological Markers; Biology; Biopsy; Brain; Cell Communication; Cell Nucleus; Cells; Characteristics; Chlorides; Cholecystokinin; Cortical Dysplasia; Data; Development; Electroencephalography; Electrophysiology (science); Epilepsy; Epileptogenesis; Frequencies; Functional disorder; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Generations; Genes; Genetic Transcription; Glutamate Decarboxylase; Heterogeneity; Hippocampus (Brain); Histologic; Histopathology; Human; Image; Impairment; Individual; Interneuron function; Interneurons; Investigation; Magnetic Resonance Imaging; Microelectrodes; Microglia; Microscopic; Molecular; Molecular Target; Neurons; Operative Surgical Procedures; Partial Epilepsies; Parvalbumins; Pathologic; Pathology; Pathway interactions; Patients; Pattern; Play; Population; Process; Pyramidal Cells; Research; Resected; Resistance; Resolution; Role; Sampling; Secondary to; Seizures; Site; Testing; Therapeutic; Tissue-Specific Gene Expression; Tissues; Transcription Alteration; base; brain tissue; calbindin; cell type; differential expression; early onset; epileptic encephalopathies; improved; insight; interest; neocortical; radiological imaging; response; sodium-potassium-chloride cotransporter 1 protein; targeted treatment; transcriptome sequencing; transcriptomics

PROJECT ABSTRACT Up to 30% of individuals with epilepsy have intractable seizures. It is unclear how mechanisms underlying seizure onset and propagation operate regionally in intractable focal epilepsy, in which tissue involvement is heterogeneous and regions play different roles in seizure initiation, propagation and resistance to spread. Better understanding is needed of the biology underlying the seizure focus versus the penumbra, where excitatory discharges appear on EEG but neuronal response is limited by intact inhibition. Imbalance of inhibitory and excitatory neuronal inputs (I/E imbalance) is a primary factor underlying seizure generation and propagation, particularly interneuron dysfunction and interneuron/pyramidal cell interaction. I/E imbalance influences seizure generation in a wide array of epilepsy pathologies; investigation of I/E imbalance and interneuron function can thereby reveal pathways common to many types and causes of epilepsy. We hypothesize that cell-type specific alterations result in I/E imbalance in brain resected during surgery for intractable focal epilepsy, and that these transcriptional changes will correlate with histopathology and regional EEG. We will test this by performing single nucleus RNA sequencing (snRNAseq) on MRI-guided electrophysiologically-localized biopsies from patients undergoing epilepsy surgery. This will provide site-specific correlation of cell type-specific transcriptomic profiles with histological, radiographic, and electrophysiologic alterations. We will sample the seizure focus and penumbra to determine the differences in cellular composition and cell type-specific expression profiles, and identify alterations involved in seizure generation and propagation. Although we will focus on interneuron profiling and I/E imbalance, snRNAseq will provide data on the diversity of cell types. In AIM 1 we will use snRNAseq to identify differential expression of genes in seizure focus vs. penumbra in human brain resected during surgery for intractable focal epilepsy, focusing on interneuron vs excitatory pyramidal cell transcriptional alterations. Microelectrode recordings will provide the spatial trajectory of a seizure and define focus and penumbra. We will identify region- and cell type-specific alterations by comparing transcriptional profiles across EEG-characterized samples. We will thereby identify molecules and pathways differentially expressed in seizure focus vs. penumbra, highlighting processes likely to be primary to seizure generation. In AIM 2 we will perform immunohistochemical analyses of top target molecules identified in Aim 1, as well as molecules previously identified to play a role in I/E imbalance in epilepsy and seizure generation. Our studies will thereby provide microscopic resolution of cellular and subcellular patterns of expression within MRI-guided electrophysiologically localized biopsies. Illuminating the regional pathophysiology of seizures in intractable focal epilepsy can provide targets for therapy.

项目摘要 多达30％的癫痫患者患有顽固性癫痫发作。目前尚不清楚癫痫发作的基础机制如何 发作和繁殖在顽固性的局灶性癫痫中在区域内作战，其中组织受累是 异质和区域在癫痫发作，传播和抗传播的抵抗中起着不同的作用。更好的 需要理解癫痫发作重点与阴茎的生物学的理解 出现在脑电图上，但神经元反应受到完整抑制的限制。抑制性不平衡和 兴奋性神经元输入（I/E失衡）是癫痫发作和传播的主要因素， 特别是内神经元功能障碍和间神经元/锥体细胞相互作用。 I/E失衡会影响癫痫发作 在各种癫痫病中产生； I/E失衡和中间功能的调查可以 因此，揭示了许多类型和癫痫原因常见的途径。我们假设细胞类型特异性 改变导致手术过程中切除的脑部切除的脑癫痫症的I/E不平衡，并且这些 转录变化将与组织病理学和区域脑电图相关。我们将通过单曲来测试这一点 来自患者的MRI引导电本生活检的RNA测序（SNRNASEQ） 接受癫痫手术。这将提供细胞类型特异性转录组轮廓的特定地点相关性 与组织学，放射学和电生理学改变。我们将采样癫痫发作的重点和 半肿，以确定细胞组成和细胞类型特异性表达谱的差异，以及 确定癫痫发作和传播涉及的变化。虽然我们将专注于中间神经 SNRNASEQ分析和I/E不平衡将提供有关细胞类型多样性的数据。在AIM 1中，我们将使用 SNRNASEQ鉴定癫痫发作焦点中基因的差异表达与切除的人脑中的阴茎 手术期间顽固性局灶性癫痫，专注于中间神经元与兴奋性金字塔细胞转录 改变。微电极记录将提供癫痫发作的空间轨迹，并定义焦点和 penumbra。我们将通过比较跨越跨的转录轮廓来识别区域和细胞类型特定的变化 EEG特征样品。因此，我们将确定在癫痫发作中差异表达的分子和途径 Focus vs. Penumbra，突出了可能是发作产生的主要过程。在AIM 2中，我们将执行 AIM 1中鉴定的顶部靶标分子的免疫组织化学分析以及先前的分子 被确定可以在癫痫和癫痫发作中发挥作用。因此，我们的研究将提供 在MRI引导的电生理学中细胞和亚细胞表达模式的微观分辨率 局部活检。阐明顽固性局灶性癫痫中癫痫发作的区域病理生理学可以提供 治疗的靶标。