The molecular basis for the translocation of fungi from blood-to-brain.

项目摘要 血脑屏障(BBB)是一种高度限制性和选择性的结构，它保护 中枢神经系统(CNS)。一些病原体，如新生隐球菌(CN)， 通过在很大程度上仍不为人所知的机制设法克服了血脑屏障。真菌孢子是 在我们的环境中流行，我们在吸入气雾化孢子后被感染 从土壤、某些类型的树木和鸽子粪便中。一旦吸入，CN的孢子就会在 肺组织，并由于其高度嗜神经的性质，传播到中枢神经系统。虽然 在中枢神经系统中观察到真菌细胞的实时报道和尸检，我们仍然 不完全了解大脑内皮细胞的分子相互作用或 调节病原体进入中枢神经系统。拟议的研究项目将涉及2 与父项目根本不同的具体目标，但重要的是，它们解决了 最重要的目标是解决定义渗透的分子机制 CN进入中枢神经系统以及随后对整体大脑健康的影响。当父母的资助集中在 EphA2-RTK作为CN进入中枢神经系统的中介作用的补充研究 本文将研究血脑屏障中的跨细胞-旁细胞串扰如何有利于 真菌病原体在血脑屏障中的渗透。AIMS将检验这样的假设：一个 穿过血脑屏障的细胞旁通路有助于CN作为一种 CN诱导的细胞骨架重塑的后果。 具体目标1将检查跨细胞作用和旁细胞通路的开放是否 在健康的大脑中正常连接或共同调节，或者如果病原体可以选择性地激活两者 小路。 具体目标2将涉及体外研究，以确定病原体是否可以严重破坏稳定 连接，并诱导伴随的细胞旁开放。拟议的研究将有一个直接的 对开发可预防神经感染的新疗法和设计新型药物的影响 旨在跨越血脑屏障将货物送到大脑的技术战略。 拟议的项目旨在作为补充，促进生物医学科学的多样性。 补充剂候选人阿米莉亚·班尼特对神经科学特别感兴趣， 神经感染，并被激励加入我的实验室，这样她就可以研究分子 驱动人类真菌病原体CN进入中枢神经系统(CNS)的机制。 尽管这一领域的发病率和死亡率很高，但仍未得到充分的研究 与真菌性脑膜脑炎有关。通过这个项目，阿米莉亚将获得 神经感染，并利用这一点来建立一个利基，使她在她的 研究生涯。该项目是专门为加强阿米莉亚对 生物医学研究，为阿米莉亚提供成功职业生涯所需的研究技能 一名独立的科学家，并为阿米莉亚提供专业的职业培训， 她希望作为一名研究人员、教育者和学术界生物医学科学的导师而蓬勃发展.

PROJECT ABSTRACT The blood-brain barrier (BBB) is a highly-restrictive and selective structure that protects the central nervous system (CNS). A few pathogens, such as Cryptococcus neoformans (Cn), have managed to overcome the BBB by mechanisms that are still largely unknown. Fungal spores are prevalent in our environment and we become infected following inhalation of aerosolized spores from soil, certain types of trees and pigeon droppings. Once inhaled, spores of Cn proliferate in lung tissue and due to their highly neurotropic nature, disseminate to the CNS. Although observations of fungal cells in the CNS have been reported in real-time and at autopsy, we still do not fully understand the molecular interactions at the brain endothelium or the pathways that mediate the movement of pathogens into the CNS. The proposed research project will address 2 specific aims that are fundamentally different from the parent project but importantly, they address the over-arching goal which is to resolve the molecular mechanisms that define the infiltration of Cn into the CNS and the subsequent effect on over-all brain health. While the parent grant focuses on the role of EphA2-RTK as a mediator of Cn infiltration into the CNS, the supplement study proposed here will examine how a transcellular-paracellular cross-talk in the BBB can favor the infiltration of fungal pathogens across the BBB. The aims will test the hypothesis that a paracellular pathway across the BBB contributes to the infiltration of Cn into the CNS as a consequence of cytoskeleton remodeling induced by Cn. Specific aim 1 will examine whether transcytosis and the opening of a paracellular path are normally linked or co-regulated in healthy brains or if pathogens can selectively activate both pathways. Specific aim 2 will involve ex-vivo studies to determine whether pathogens can destabilize tight junctions and induce a concomitant paracellular opening. The proposed studies will have a direct impact on developing new therapeutics that could prevent neuroinfections and on designing novel strategies for technologies geared toward crossing the BBB to deliver cargo to the brain. The proposed project is intended as a supplement to promote diversity in the biomedical sciences. The supplement candidate, Amelia Bennett, has a strong interest in neuroscience specifically, neuroinfections, and was motivated to join my lab so that she could study the molecular mechanisms that drive a human fungal pathogen, Cn, into the central nervous system (CNS). This area of research is understudied despite the significant morbidity and mortality that is associated with fungal meningoencephalitis. Through this project Amelia will acquire expertise in neuroinfections and use this to establish a niche that will allow her to be competitive in her research career. The project has been specifically designed to enhance Amelia’s participation in biomedical research, to provide Amelia with the research skills needed for a successful career as an independent scientist and to provide Amelia with the professional career training that will allow her to thrive as a researcher, educator and mentor in biomedical sciences in academia.