CAREER: Rational design of a biomimetic nanomaterial library to probe mechanisms behind virus-induced immunopathology

PI：Scott，Evan A.Proposal number：1453576病毒是复杂的纳米级结构，进化出不同的策略来影响免疫系统以逃避检测并延长其在宿主中的停留时间。由此产生的病毒相关的病理生理学伴随着良好的炎性免疫反应，旨在纠正被破坏的体内平衡。病毒的形态、表面化学和细胞靶标指导着这种炎症，在许多情况下，这种炎症会变得失调，导致一系列比原始病毒感染本身更有害的病理。因此，了解可以诱导特定免疫反应的特定炎症触发因素对于识别和治疗病毒诱导的病理以及影响包括动脉粥样硬化甚至季节性过敏在内的广泛的炎症驱动的疾病状态是至关重要的。不幸的是，导致这种有害的免疫失调的具体机制往往非常复杂，很难分离出来。生物医学工程(BME)和纳米技术的最新进展使人们能够对合成纳米结构的物理和化学性质进行前所未有的控制，这种结构可以被设计成模仿病毒等纳米级病原体的感染机制。因此，这项提议的研究目标是设计模仿病毒的纳米结构，作为工具来研究病毒如何产生失调免疫反应的特定假说。随着工程学方法解决免疫学问题变得越来越普遍，对免疫学和病毒学高度专业领域感兴趣的未来学生和研究人员必须被鼓励在BME领域从事职业。因此，这项提案的教育目标是实施一项多管齐下的高中BME计划，让学生和他们的父母接触到在BME工作所带来的各种机会。这项建议旨在通过直接展示基于工程的技术和原理的好处，将BME扩展到生物科学的新的专业领域。病毒免疫病理学研究的一个关键需要是一种可定制的原位方法，以探索特定的病毒结构、表面化学和生物分布如何导致病毒诱导的免疫系统调节失调。PI将合理地设计一个模拟病毒的聚合物纳米载体(NCS)文库，以研究触发模型免疫功能紊乱噬血细胞淋巴组织细胞增多症(HLH)的基本生化和细胞机制。在淋巴细胞性脉络膜脑膜炎病毒(LCMV)感染后的几种小鼠模型中可以可控地诱导HLH。由于聚合物是惰性的，不会引发炎症反应，NCS可以被认为是“白板”，其中将结合各种分子和免疫刺激剂，以控制特定免疫细胞的运输和激活。诱导HLH的基本炎症机制尚不清楚，因此，将LCMV内的各种炎症成分，如其糖蛋白包膜或RNA，划分为模拟病毒的生物分布和细胞内降解机制的单独的NC，将简化这一系统。NCs是由嵌段共聚物组装而成的，可以被工程化为泡状和丝状纳米结构，分别针对不同的和受限的炎症免疫细胞亚群。囊泡将通过诱导受控的全身性细胞因子表达和效应器T细胞激活来探索导致高促黄体生成素的假想生化机制。这些细丝将研究浆细胞样树突状细胞的特定细胞作用，浆细胞样树突状细胞是一种炎性免疫细胞群，对抗LCMV的免疫反应至关重要。通过使用可控制的人工诱导感染，将确定与LCMV相关的关键炎症机制或其组合，从而导致HLH。这项工作将展示合理设计的纳米材料的使用，以增强对特定炎症触发因素如何有助于免疫病理学的理解。此外，在这些研究中收集的数据将提供对诱导特定和受控免疫反应所需的基本设计标准的洞察。

PI: Scott, Evan A.Proposal Number: 1453576Viruses are intricate nanoscale structures that have evolved diverse strategies of influencing the immune system to evade detection and prolong residence within their hosts. The resulting virus-related pathophysiology is accompanied by well-intended inflammatory immune responses aimed at rectifying a disrupted homeostasis. The morphology, surface chemistry, and cellular targets of viruses directs this inflammation, which in many instances becomes dysregulated, resulting in a range of pathologies that can be more harmful than the original viral infection itself. Understanding the specific inflammatory triggers that can induce a particular immunological response is therefore essential to the identification and treatment of virus-induced pathologies as well as impact a broad range of inflammation-driven disease states including atherosclerosis and even seasonal allergies. Unfortunately, the specific mechanisms that are responsible for such harmful immune dysregulations are often prohibitively complex and difficult to isolate. Recent advances in biomedical engineering (BME) and nanotechnology now permit unprecedented control over the physical and chemical properties of synthetic nanostructures, which can be designed to mimic the mechanisms of infection utilized by nanoscale pathogens such as viruses. The research objective of this proposal is therefore to engineer virus-mimicking nanostructures that will serve as tools to investigate specific hypotheses of how viruses can generate dysregulated immune responses. As engineering approaches to immunological questions become more commonplace, future students and researchers interested in highly specialized areas of immunology and virology must be encouraged to pursue careers in the field of BME. The educational objective of this proposal therefore implements a multi-pronged high school BME program that exposes students, as well as their parents, to the diverse opportunities made accessible by a career in BME. This proposal aims to expand BME into new specialized fields of biological science by directly demonstrating the benefits of engineering-based techniques and principles.A key need in the study of viral immunopathology is a customizable, in situ method to probe how specific viral structures, surface chemistries, and biodistributions contribute to virus-induced dysregulation of the immune system. The PI will rationally design a library of virus-mimicking polymeric nanocarriers (NCs) to investigate the basic biochemical and cellular mechanisms that trigger the model immunodysregulatory disorder hemophagocytic lymphohistiocytosis (HLH). HLH can be controllably induced in several mouse models following infection by the lymphocytic choriomeningitic virus (LCMV). Since the polymers are inert and do not elicit inflammatory responses, the NCs can be considered "blank slates" in which a diverse range of molecules and immunostimulants will be incorporated for controlled transport to and activation of specific immune cells. The essential inflammatory mechanisms that induce HLH are poorly understood, and so this system will be simplified by dividing the various inflammatory components within LCMV, such as its glycoprotein envelope or RNA, into separate NCs that mimic the biodistribution and mechanisms of intracellular degradation of the virus. NCs are assembled from block copolymers, and can be engineered into vesicular and filamentous nanostructures, which respectively target diverse and restricted subsets of inflammatory immune cells. Vesicles will probe hypothesized biochemical mechanisms responsible for HLH by inducing controlled systemic cytokine expression and effector T cell activation. The filaments will investigate the specific cellular role of plasmacytoid dendritic cells, which are an inflammatory immune cell population essential to immune responses against LCMV. Through the use of controlled synthetically induced infections, the key LCMV-related inflammatory mechanisms or combinations thereof responsible for HLH will be identified. This work will demonstrate the use of rationally designed nanomaterials to enhance the understanding of how specific inflammatory triggers contribute to immunopathology. Furthermore, data gathered in these studies will provide insight into essential design criteria required for eliciting specific and controlled immunological responses.