Volumetric optical connectome microscopy of human cerebellar circuitry

项目概要/摘要 寻求K99/R 00独立之路奖的目标是建立自己作为一个独立的 首席研究员，研究正常人和大脑中大脑回路的结构-功能关系 紊乱拟议的项目，由需要了解人类大脑的高分辨率驱动， 高通量工具和我在神经成像生物医学光学方面的丰富经验，旨在建立一个 多功能的工具来重建人类小脑的电路和神经元结构， 小脑退行性疾病的破坏性影响，并与MRI模型联合收割机，以寻求新的生物标志物 这可能会影响临床评估。 尽管自圣地亚哥·拉蒙·卡哈尔在1989年的开创性工作以来，光学显微镜取得了巨大的进步， 绘制轴突束，我们对80-100亿神经元如何连接在一起形成复杂的 人类大脑的功能仍然有限。目前，还没有体积显微镜技术可以映射 高度完整的人脑电路和结构。在这里，我建议开发一个体积光学 连接组显微镜（VOCM），用于重建人类小脑前所未有的分辨率， 尺度，并从全局的角度映射连接和神经元架构。VOCM基于 偏振敏感光学相干断层扫描仪和振动切片机对大规模离体大脑进行成像 以微米级的分辨率。重要的是，该技术允许体积重建， 超高精度，无组织变形;因此克服了所有组织学的100年挑战 基于追踪长纤维束和检查人脑复杂皮质折叠的方法。的 VOCM产生的高质量数据将被拟合到MRI模型中，以构建一个超高分辨率的 人类小脑，以提供在当前MRI工具中不可用的解剖标记。 通过应用VOCM，该项目进一步探索小脑疾病的3D病理模式。多系统 小脑萎缩型（MSA-C）是一种致命的神经退行性疾病， 脑干萎缩尽管其发病率很低，但MSA-C与其他疾病有共同的表型特征。 神经系统疾病。研究MSA-C的神经解剖学基础和病理轨迹， 促进我们对小脑疾病和小脑受累疾病的影响的理解。尤其是， 该项目将描述与神经变性相关的结构和电路中断， MSA-C然后，我们将使用高分辨率的体外数据集进行体内预测，并允许MRI 这是不可能的评估，否则。 拟议的研究在马萨诸塞州总医院（MGH）的Martinos中心进行，该中心是一家 一个理想的环境，发展尖端的生物医学技术，并与大型社区互动， 具有多学科背景的专家。我已经组建了一个强大的指导团队：布鲁斯菲舍尔博士， Martinos中心的计算核心; Martinos中心光学部主任大卫博厄斯博士; 以及麻省总医院共济失调科主任杰里米·施曼博士我将利用MRI建模方面的正式培训 小脑相关的脑疾病和神经病理学。神经解剖学的课程， 神经生物学和中枢神经系统疾病补充了我的生物医学光学背景，并提高了我的 重要神经科学问题的知识。建议的培训和研究将使我准备好 必要的技能，新的工具和有趣的数据，以启动一个独立的研究计划和写作进一步 R 00阶段完成后的研究赠款。我希望这项研究能极大地促进我们的 当前脑科学知识，对临床革命产生影响，并倡导公众意识， 更多人群的大脑健康。

Project Summary/Abstract The goal in seeking a K99/R00 Pathway to Independence Award is to establish myself as an independent principal investigator to study the structural-functional relationship of the brain circuitry in normal and brain disorders. The proposed project, driven by the need for understanding the human brain with high-resolution high-throughput tools and my extensive experience in biomedical optics for neuroimaging, aims to establish a versatile tool to reconstruct the circuitry and neuronal architecture in human cerebellum, understand the disruptive impacts of cerebellar degenerative disease, and combine with MRI models to seek novel biomarkers that will potentially influence the clinical assessment. Despite the tremendous advances of light microscopy since Santiago Ramón y Cajal's pioneering work in drafting axonal tracts, our knowledge on how the 80-100 billions of neurons connect together to form complex functions in human brain is still limited. Presently, there is no volumetric microscopy technique that can map the circuitry and architecture of human brain with high integrity. Here I propose to develop a volumetric optical connectome microscopy (VOCM), for reconstructing the human cerebellum with unprecedented resolution and scales, and mapping the connectivity and neuronal architectures from a global perspective. VOCM is based on a polarization sensitive optical coherence tomography and a vibratome slicer to image large-scale ex vivo brain at a micrometer-scale resolution. Importantly, this technology allows volumetric reconstruction preserving an ultra-high accuracy without tissue distortions; therefore overcomes the 100 years challenge of all histology based methods in tracing long fiber tracts and inspecting sophisticated cortical folding in the human brain. The high-quality data generated by VOCM will be fit into MRI models to construct an ultra-high resolution atlas of human cerebellum to provide anatomical labels that are not available in current MRI tools. By applying VOCM, the project further explores 3D pathological patterns of cerebellar disorder. Multiple system atrophy cerebellar type (MSA-C) is a fatal neurodegenerative disease manifested by severe cerebellar and brainstem atrophy. Despite its rare incidence, MSA-C shares common phenotypic characteristics with other neurological diseases. Studying the neuroanatomical substrates and pathological trajectory of MSA-C could advance our understanding of the impact of cerebellar disorders and cerebellar affected diseases. Particularly, the project will characterize the architecture and circuitry disruptions associated with neurodegeneration in MSA-C. We will then use the high-resolution ex vivo dataset to make predictions in vivo, and allow an MRI assessment that would not be possible otherwise. The proposed research is conducted at Martinos Center, Massachusetts General Hospital (MGH), which is an ideal environment developing cutting edge biomedical technologies and interacting with a large community of experts with multidisciplinary background. I have formed a strong mentoring team: Dr. Bruce Fischl, director of the Computational Core at Martinos Center; Dr. David Boas, director of the Optics Division at Martinos Center; and Dr. Jeremy Schmahmann, director of the MGH Ataxia Unit. I will leverage formal trainings in MRI modeling and analysis, cerebellar related brain diseases and neuropathology. The coursework on neuroanatomy, neurobiology and central nervous diseases complement my biomedical optics background and advance my knowledge on important neuroscience questions. The proposed trainings and research will prepare me with necessary skills, new tools, and intriguing data to launch an independent research program and writing further research grants after the completion of the R00 phase. I expect that the research will dramatically advance our current knowledge on brain science, have an impact on clinical revolutions, and advocate public awareness of brain health in greater populations.