Optogenetic approaches to study post-stroke recovery mechanisms

研究中风后恢复机制的光遗传学方法

• 批准号: 10530685
• 负责人: GARY K STEINBERG
• 金额: $ 61.33万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10530685
• 关键词: Address; Affect; Animals; Area; Automobile Driving; Biological; Brain; Brain Diseases; Brain Mapping; Brain region; Calcium; Cause of Death; Cell Count; Characteristics; Cholesterol; Clinical Research; Corpus striatum structure; Data; Event; Female; Frequencies; Functional Magnetic Resonance Imaging; Future; G Protein-Coupled Receptor Signaling; Genes; Goals; Grant; High-Throughput RNA Sequencing; Image; Imaging Techniques; Individual; Infarction; Intervention; Location; Maps; Medical; Metabolism; Microscope; Molecular; Molecular Profiling; Molecular Target; Motor Cortex; Mus; Neurons; Parvalbumins; Pathway interactions; Population; Recovery; Recovery of Function; Reporting; Resolution; Role; Sex Differences; Site; Somatosensory Cortex; Stains; Stroke; Techniques; Testing; Thalamic structure; Therapeutic; Therapeutic Intervention; Time; Translating; Validation; behavior test; brain remodeling; calmodulin-dependent protein kinase II; cell type; cholesterol biosynthesis; design; disability; effective therapy; excitatory neuron; functional improvement; imaging system; improved; inhibitory neuron; insight; male; multimodality; nervous system disorder; neural; neural circuit; neuroimaging; neuroregulation; new therapeutic target; optogenetics; partial recovery; portability; post stroke; promoter; sex; stroke outcome; stroke recovery; tool; transcriptome; transcriptome sequencing; transcriptomic profiling

PROJECT SUMMARY Stroke is the leading cause of death with very limited treatment options. This devastating neurological disease is increasingly viewed as a disease of brain connectivity as a damaged stroke area can affect both local and connected brain regions, causing disruptions in neuronal activity and metabolism network-wide. Recovery of lost function can occur after stroke and is attributed to brain remodeling in areas adjacent to or connected to the infarct. In this proposal, we aim to investigate the role of key brain circuits in post-stroke recovery at the functional, cellular and molecular level, using optogenetics, advanced live imaging and high throughput RNA sequencing techniques. Previously our lab has demonstrated that selective optogenetic neuronal stimulation in the ipsilesional motor cortex (iM1) can activate plasticity mechanisms and promote recovery. Recently we have employed the optogenetic functional MRI technique to systematically map brain-wide changes in neural circuits after stroke. We have identified key circuits altered by stroke and demonstrated two key circuits restored by iM1 stimulations. Our map data also revealed two candidate circuits that were not restored by iM1 stimulations, suggesting that greater recovery could be achieved if we can rescue these circuits by directly stimulating them. In this proposal we aim to investigate key neural circuits we identified from our activation maps and elucidate their role in post-stroke recovery. In Aim1 we will use circuit-specific optogenetic tools and functional behavior tests to interrogate the role of key circuits in post-stroke recovery. This aim will address whether these circuits have beneficial or maladaptive role during post-stroke recovery. In Aim2 we will examine cellular resolution of real-time neuronal activity dynamics in key circuits after stroke using a portable live calcium imaging system. This will elucidate the neural activity dynamics (excitatory and inhibitory) of key circuits at the cellular level, allowing us to identify the temporal profile and the key neuronal populations altered by stroke, and how iM1 stimulations affect these characteristics to enhance recovery. In Aim3 we will investigate the transcriptome of key circuit areas using RNAseq, in order to identify key molecular targets and pathways altered by stroke and by iM1 stimulations. Preliminary RNAseq analysis revealed distinct pathways altered by iM1 stimulations. We aim to perform RNAseq in multiple regions including iM1 (stimulation site) and ipsilesional thalamus (iM1- connected region) to elucidate whether similar pathways are involved, and if we can identify a common molecular signature that drive recovery. We will also perform RNAseq in both sexes in order to ascertain any sex-specific differences that may be present in post-stroke recovery. Together these results will 1) advance the understanding of neural circuit dynamics during post-stroke recovery; and 2) identify key neural circuits/cell types/molecular targets and optimal time window for designing brain stimulation strategies and other therapeutic interventions in future clinical studies.

项目摘要 中风是死亡的主要原因，治疗方案非常有限。这种毁灭性的神经疾病 越来越被视为脑连通性疾病，因为中风区受损会影响局部和 连接的大脑区域，导致神经元活性和范围内代谢的干扰。恢复 中风后可能发生丢失的功能，并归因于邻近或连接的区域的脑重塑 梗塞。在此提案中，我们旨在调查关键大脑电路在冲程后恢复中的作用 功能性，细胞和分子水平，使用光遗传学，先进的活成像和高吞吐量RNA 测序技术。以前我们的实验室证明了选择性的光遗传学神经元刺激 iPsiles运动皮层（IM1）可以激活可塑性机制并促进恢复。最近我们有 采用光遗传学功能MRI技术来系统地绘制神经回路的脑部变化 中风后。我们已经确定了通过中风更改的关键电路，并证明了两个主要电路。 IM1刺激。我们的地图数据还显示了两个未被IM1刺激恢复的候选电路， 如果我们可以通过直接刺激这些电路来挽救这些电路，则表明可以实现更大的恢复。 在此提案中，我们旨在调查我们从激活图中确定的关键神经回路并阐明 他们在势后恢复中的作用。在AIM1中，我们将使用特定电路的光遗传学工具和功能行为 测试以询问关键电路在冲程后恢复中的作用。这个目标将解决这些电路是否 在击球后恢复期间，具有有益或不良适应的作用。在AIM2中，我们将检查的细胞分辨率 使用便携式实时钙成像系统，中风后关键回路中的实时神经元活性动力学。 这将阐明细胞水平关键回路的神经活动动态（兴奋性和抑制性）， 允许我们识别中风改变的时间概况和关键神经元种群，以及IM1如何 刺激会影响这些特征以增强恢复。在AIM3中，我们将研究 使用RNASEQ的关键电路区域，以识别中风和 通过IM1刺激。初步RNASEQ分析显示，IM1刺激改变了不同的途径。我们 旨在在包括IM1（刺激位点）和ipsiles thalamus（IM1-）在内的多个地区执行RNASEQ 连接区域）以阐明是否涉及类似的途径，以及是否可以识别一个共同的途径 驱动恢复的分子签名。我们还将在两个性别中执行rnaseq，以确定任何 冲程后恢复中可能存在性别差异。这些结果将共同提前 对冲程后恢复期间神经电路动态的了解； 2）确定关键的神经回路/细胞 类型/分子靶标和最佳的时间窗口，用于设计大脑刺激策略和其他 将来的临床研究中的治疗干预措施。