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，它保护脑细胞的正常功能。 中枢神经系统（CNS）。一些病原体，如新型隐球菌（Cn）， 设法克服血脑屏障的机制，仍然在很大程度上是未知的。真菌孢子是 在我们的环境中流行，我们在吸入雾化的孢子后被感染 从土壤，某些种类的树木和鸽子粪便中提取一旦被吸入，Cn的孢子在体内增殖， 肺组织，并由于其高度嗜神经性，传播到中枢神经系统。虽然 CNS中真菌细胞的观察已经被实时报道，在尸检中，我们仍然 并不完全了解脑内皮细胞的分子相互作用或 介导病原体进入CNS的运动。该研究项目将解决2 与母项目有根本不同的具体目标，但重要的是， 过度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个大脑健康的影响。虽然父母补助金的重点是 关于EphA 2-RTK作为Cn浸润到CNS中的介质的作用，补充研究 这里提出的将研究如何在BBB跨细胞旁串扰可以有利于 真菌病原体穿过血脑屏障的浸润。这些目标将检验一个假设， 穿过BBB的细胞旁途径有助于Cn作为一种免疫调节剂浸润到CNS中。 结果由Cn诱导的细胞骨架重塑。 具体目标1将检查转胞吞作用和细胞旁通路的打开是否是 正常连接或共同调节在健康的大脑，或者如果病原体可以选择性地激活两者 途径。 具体目标2将涉及离体研究，以确定病原体是否可以破坏紧密的 连接并诱导伴随的细胞旁开放。拟议的研究将直接 对开发预防神经感染的新疗法和设计新的 这些技术的战略是为了跨越血脑屏障向大脑运送货物。 拟议的项目旨在作为促进生物医学科学多样性的补充。 补充候选人，阿米莉亚班尼特，有一个强烈的兴趣，特别是神经科学， 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。 这一领域的研究是不足的，尽管显着的发病率和死亡率， 与真菌性脑膜脑炎有关通过这个项目，Amelia将获得以下方面的专业知识： 神经感染，并利用这一点来建立一个利基，这将使她在她的竞争力， 研究生涯。该项目是专门设计来提高阿米莉亚的参与， 生物医学研究，为阿米莉亚提供一个成功的职业生涯所需的研究技能， 一个独立的科学家，并为阿米莉亚提供专业的职业培训， 她在学术界的生物医学科学作为一个研究人员，教育家和导师茁壮成长。

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.