Deciphering the genomic mechanisms underlying the physiology of human brain stimulation

破译人脑刺激生理学背后的基因组机制

• 批准号: 10559426
• 负责人: Genevieve Konopka
• 金额: $ 379.03万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10559426
• 关键词: Acute; Address; Affect; Animal Model; Area; Brain; Brain region; Cell Nucleus; Cells; Chromatin; Chronic; Cognition; Cognitive; Complex; Data; Development; Electric Stimulation; Electrophysiology (science); Epigenetic Process; Excision; Exhibits; Exposure to; Frequencies; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Transcription; Genomics; Human; Immediate-Early Genes; In Vitro; Influentials; Infrastructure; Ion Channel; Knowledge; Lateral; Link; Lobectomy; Measures; Memory; Methods; Modeling; Movement Disorders; Neurons; Operative Surgical Procedures; Patients; Pattern; Performance; Physiological; Physiology; Process; Publishing; Research; Resected; Resolution; Role; Series; Signal Transduction; Slice; Small Nuclear RNA; Specimen; Structure of middle temporal gyrus; Synaptic plasticity; System; Techniques; Temporal Lobe; Testing; Time; Tissues; Work; XCL1 gene; base; brain tissue; cell type; chromatin remodeling; clinical application; clinically relevant; detection method; epigenomics; expectation; experience; experimental study; flexibility; gene network; human subject; human tissue; in vivo; innovation; memory process; multi-electrode arrays; neural circuit; neurophysiology; neuroregulation; novel; predictive modeling; relating to nervous system; response; success; tissue culture; tissue preparation; tissue processing; transcription factor; transcriptome sequencing; transcriptomics

The underlying mechanisms of brain stimulation in humans are poorly understood, especially at the level of gene expression. To address this gap in knowledge, we propose a series of three experiments that take advantage of the opportunity to obtain high-quality human neural tissue from neurosurgical patients in order to measure the impact of brain stimulation on gene expression. Our experiments will generate data to explicate changes at the level of gene expression that underlie brain circuit changes elicited by stimulation. Our study team has seven years of experience analyzing gene expression using an established pipeline for studying human cortical tissue from neurosurgical patients, including application of cutting-edge methods for measuring gene expression. These methods include single nuclei RNA-sequencing (snRNA-seq) and the exciting addition of single nuclei ATAC-sequencing (snATAC-seq) to understand stimulation-related changes in transcription factors and chromatin remodeling. Our hypotheses regarding specific gene classes were developed from our published data correlating gene expression changes with neurophysiological signatures (brain oscillations) linked with successful memory formation. In this proposal, our experiments address the complex problem of how stimulation alters neural circuits using three complementary approaches. First, we will use direct cortical stimulation in vivo immediately prior to resection of brain tissue in temporal lobectomy patients, followed by gene expression analysis. Our plans are supported by preliminary data showing differences in expression of immediate early genes (IEGs) following cortical stimulation, in line with predictions drawn from animal models. Second, we will build on techniques we have implemented for culture of human neural tissue (from neurosurgical patients) to measure gene expression changes elicited by chronic ex vivo stimulation. This experiment will elucidate the temporal dynamics of gene expression in the setting of stimulation, including transcription factor changes, using our experience with time series modeling of gene information. Finally, we will use multi-electrode arrays (MEAs) to measure the impact of ex vivo stimulation on networks of co-firing neurons, directly testing models of stimulation-induced changes in local circuits. We will connect these electrophysiological measures with gene expression changes elicited by stimulation. This experiment builds on our published work studying network activity in single unit recordings in humans, as well as our preliminary data demonstrating the ability to record electrophysiological signals from human neural tissue in culture. The stimulation parameters were developed to be aligned across in vivo and in vitro experiments to facilitate comparison. Taken together, our experiments will provide ground-breaking data elucidating the genetic underpinnings of how brain stimulation elicits neuromodulation. The experience of research team and proven record in publishing data using neurosurgical tissue specimens supports our expectations of success.

人类大脑刺激的潜在机制知之甚少，特别是在基因表达水平上。为了解决这一知识差距，我们提出了一系列的三个实验，利用机会从神经外科患者获得高质量的人类神经组织，以测量脑刺激对基因表达的影响。我们的实验将产生数据来解释基因表达水平的变化，这些变化是刺激引起的脑回路变化的基础。我们的研究团队拥有七年的经验，使用已建立的管道研究神经外科患者的人类皮质组织，包括应用尖端方法测量基因表达。这些方法包括单核RNA测序（snRNA-seq）和令人兴奋的单核ATAC测序（snATAC-seq），以了解转录因子和染色质重塑中刺激相关的变化。我们关于特定基因类别的假设是从我们发表的数据中发展出来的，这些数据将基因表达变化与成功记忆形成相关的神经生理学特征（脑振荡）相关联。在这个提议中，我们的实验解决了刺激如何使用三种互补方法改变神经回路的复杂问题。首先，我们将在颞叶切除术患者的脑组织切除前立即使用直接皮质刺激，然后进行基因表达分析。我们的计划得到了初步数据的支持，这些数据显示了皮层刺激后即刻早期基因（IEG）表达的差异，与动物模型的预测一致。其次，我们将建立在我们已经实施的人类神经组织（来自神经外科患者）培养技术的基础上，以测量慢性离体刺激引起的基因表达变化。本实验将阐明的时间动力学的基因表达的设置刺激，包括转录因子的变化，使用我们的经验与时间序列建模的基因信息。最后，我们将使用多电极阵列（MEA）来测量离体刺激对共激发神经元网络的影响，直接测试局部电路中刺激诱导变化的模型。我们将这些电生理措施与刺激引起的基因表达变化。这项实验建立在我们发表的研究人类单个单位记录网络活动的工作基础上，以及我们的初步数据，这些数据证明了在培养中记录人类神经组织电生理信号的能力。开发的刺激参数在体内和体外实验中保持一致，以便于比较。总之，我们的实验将提供突破性的数据，阐明大脑刺激如何增强神经调节的遗传基础。研究团队的经验和使用神经外科组织标本发表数据的可靠记录支持我们对成功的预期。