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

基本信息

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
项目类别：
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

项目摘要

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.

人类脑刺激的潜在机制知之甚少，尤其是在基因表达水平上。为了解决知识的这一差距，我们提出了一系列三个实验，这些实验利用了从神经外科患者那里获得高质量人类神经组织的机会，以衡量脑刺激对基因表达的影响。我们的实验将生成数据，以阐明基因表达水平的变化，而基因表达的水平是脑电路的构成的刺激变化。我们的研究团队有7年的经验，使用已建立的管道来分析基因表达，用于研究神经外科患者的人皮质组织，包括应用尖端方法来测量基因表达。这些方法包括单核RNA - 序列（SNRNA-SEQ）以及激发单核ATAC-Sequencing（SNATAC-SEQ）的激动人心的添加，以了解转录因子和染色质重塑的刺激相关的变化。我们关于特定基因类别的假设是从我们公开的数据中提出的，该数据将基因表达的变化与神经生理学特征（脑振荡）相关联，与成功的记忆形成有关。在此提案中，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中切除脑组织之前立即使用直接的皮质刺激，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，显示皮质刺激后立即产生基因（IEG）表达的差异，这与动物模型的预测一致。其次，我们将建立在为人类神经组织培养（从神经外科患者中）实施的技术，以测量慢性外体外刺激引起的基因表达变化。该实验将使用我们的时间序列模型的经验来阐明基因表达的时间动力学，包括转录因子变化。最后，我们将使用多电极阵列（MEA）来衡量离体刺激对共接触神经元网络的影响，直接测试刺激引起的局部电路变化的模型。我们将将这些电生理测度与刺激引起的基因表达变化联系起来。该实验基于我们已发表的工作研究人类单位记录中的网络活动，以及我们的初步数据，证明了能够记录培养物中人类神经组织的电生理信号的能力。开发了刺激参数，以对齐在体内和体外实验中，以促进比较。综上所述，我们的实验将提供开创性的数据，以阐明脑刺激如何引起神经调节的遗传基础。研究团队的经验和使用神经组织标本发布数据的经验记录支持我们对成功的期望。