Inflammation mediated evolving changes in synapse structural integrity and dynamics in vivo in a mouse model of multiple sclerosis

这是马萨诸塞州综合医院的神经科医生丽贝卡·吉拉尼申请K08奖项。她在神经发育障碍和中风的啮齿动物模型中接受过严格的神经可塑性方面的培训。在最初的博士后工作经验中，她获得了体内双光子成像技术方面的专业知识，该技术致力于一个项目，以确定视觉皮质电路连接的基本原理。她现在的目标是利用这些最先进的技术对神经元和突触进行体内双光子成像，并将它们应用于多发性硬化症的啮齿动物临床前模型中的神经退化研究，这是她的临床兴趣领域。这个职业发展奖的目标是为她提供从生理回路布线和可塑性研究过渡到神经炎症疾病模型的独立研究的技能，培训如下：1)神经变性、神经免疫学和体内双光子成像专家的多学科指导；2)多发性硬化症的实验性自身免疫性脑脊髓炎小鼠模型的方法学；3)免疫学和生物统计学方法学的教学培训；以及4)沟通和领导职业发展。她的导师Brian Bacskai是神经退行性疾病的体内双光子成像专家，她的共同导师Elly Nedivi是神经元和突触结构的体内双光子成像专家。她的顾问包括神经变性专家鲁道夫·坦齐，以及神经免疫学专家弗朗西斯科·昆塔纳和斯塔西·比尔博。虽然有大量的文献和正在进行的工作来定义多发性硬化症病理基础的免疫机制，但一个关键的需求领域是定义多发性硬化症过程中神经元和突触功能障碍和退化的特定机制。这些神经退行性改变是多发性硬化症残疾的关键决定因素。多发性硬化症早期的炎症很可能不会立即导致突触丢失。相反，突触结构完整性和动力学的一系列演变变化更有可能是突触丧失的最终结果。在目标1中，吉拉尼博士将利用体内最先进的双光子成像技术，在多发性硬化症模型中纵向监测活体小鼠皮质锥体神经元上所有兴奋性和抑制性连接。在目标2中，吉拉尼博士将使用细胞结构的可逆扩张和超分辨率成像的新方法，通过观察目标1中活体动物的神经元和突触图像，分析与多发性硬化症的突触病理有关的重要突触蛋白和炎症因子的存在。她提出的研究将提供一幅全面的图景，展示在多发性硬化症小鼠模型中，皮质锥体细胞整个树状突触上的不同突触群体如何在8周内发生动态变化。这项研究将作为第四年R01申请K08奖项的基础。

This is an application for a K08 award for Dr. Rebecca Gillani, a neurologist at the Massachusetts General Hospital. She has rigorous prior training in neuroplasticity in rodent models of a neurodevelopmental disorder and stroke. In her initial post-doctoral experience, she has gained expertise in in vivo two-photon imaging technology working on a project to define basic principles of circuit wiring in the visual cortex. She now aims to take these state-of-the-art techniques for in vivo two-photon imaging of neurons and synapses and apply them to the study of neurodegeneration in rodent preclinical models of multiple sclerosis, her area of clinical interest. The goal of this career development award is to provide her the skills to make the transition from the study of physiologic circuit wiring and plasticity, to the independent study of disease models of neuroinflammation with training as follows: 1) Multi-disciplinary mentoring from experts in neurodegeneration, neuroimmunology, and in vivo two-photon imaging; 2) methodology for experimental autoimmune encephalomyelitis mouse model of multiple sclerosis; 3) didactic training in immunology and biostatistical methodology; and 4) communication and leadership career development. Her mentor Brian Bacskai is expert in in vivo two-photon imaging in neurodegenerative diseases, and her co-mentor Elly Nedivi is expert in in vivo two-photon imaging of neuron and synapse structure. Her advisors include experts in neurodegeneration, Rudolph Tanzi, and experts in neuroimmunology, Francisco Quintana and Staci Bilbo. While there is an immense literature and ongoing effort to define the immune mechanisms underlying multiple sclerosis pathology, a critical area of unmet need is defining the specific mechanisms of neuronal and synaptic dysfunction and degeneration during the course of multiple sclerosis. These neurodegenerative changes are a key determinant of disability in multiple sclerosis. It is likely that inflammation in early multiple sclerosis does not immediately lead to synapse loss. Instead there is more likely a series of evolving changes in synapse structural integrity and dynamics with the eventual outcome of synapse loss. In Aim 1, Dr. Gillani will leverage state-of-the- art in vivo two-photon imaging technologies for longitudinal monitoring of all the excitatory and inhibitory connections onto individual cortical pyramidal neurons in a living mouse in a model of multiple sclerosis. In Aim 2, Dr. Gillani will use new methods for reversible expansion and super-resolution imaging of cellular architecture to analyze for the presence of important synaptic proteins and inflammatory factors implicated in synaptic pathology in multiple sclerosis, by observing the same neurons and synapses imaged in live animals in Aim 1. The studies she proposes will provide a comprehensive picture of how diverse populations of synapses across the entire dendritic tree of cortical pyramidal cells undergo dynamic changes over 8 weeks in a mouse model of multiple sclerosis. This research will be the basis of a R01 application in year 4 of this proposed K08 award.