Engineering a human brain organoid-based platform to study neurotropic viruses

设计一个基于人脑类器官的平台来研究嗜神经病毒

• 批准号: 9461004
• 负责人: Guo-li Ming
• 金额: $ 154.33万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9461004
• 关键词: Ache; Address; Adherent Culture; Adopted; Adult; Affect; Americas; Animals; Arthralgia; Basic Science; Bioinformatics; Biological; Biological Assay; Biology; Biomedical Engineering; Brain; Cell model; Cell physiology; Cells; Centers for Disease Control and Prevention (U.S.); Central Nervous System Infections; Central Nervous System Viral Diseases; Cerebrum; Coculture Techniques; Collaborations; Communicable Diseases; Communities; Complex; Custom; Data; Decision Making; Development; Diagnostic; Diarrhea; Disease; Disease Outbreaks; Encephalitis; Engineering; Epidemic; Exanthema; Fibroblasts; Flavivirus; Flavivirus Infections; Fostering; Future; Generations; Goals; Headache; Human; Immune response; Infectious Diseases Research; Institutes; Intervention; Laboratories; Lead; Logistics; Meningitis; Methodology; Methods; Microcephaly; Mitotic; Modeling; Molecular; Names; Nervous system structure; Neuraxis; Neurologic; Neurological outcome; Neurons; Organ; Organoids; Pathology; Pharmaceutical Preparations; Physiology; Population; Pregnant Women; Proliferating; Property; Protocols documentation; Research; Research Personnel; Research Project Grants; Resources; Site; Skin; Somatic Cell; Source; Standardization; Structure; System; Technology; Testing; Therapeutic Intervention; Time; Training; Translational Research; Treatment Efficacy; Tropism; Validation; Viral; Virus; Vomiting; West Nile viral infection; West Nile virus; ZIKV infection; Zika Virus; base; cell type; cost; data resource; data sharing; design; drug development; drug testing; fetal; flexibility; functional outcomes; global health; human stem cells; human tissue; in vitro Model; induced pluripotent stem cell; infectious disease model; member; molecular pathology; nerve stem cell; neurodevelopment; neurotropic; neurotropic virus; new technology; organ growth; programs; response; scale up; screening; stem cell biology; stem cell differentiation; success; technology development; therapeutic development; therapeutic evaluation; three dimensional cell culture; translational study; virology; virus pathogenesis

SUMMARY – Overview Modeling of infectious diseases that affect the human central nervous system (CNS), such as those associated with Zika virus (ZIKV) and West Nile virus (WNV), has been challenging due to the inaccessibility of the relevant cell types. Reprogramming human somatic cells, such as skin fibroblasts, into induced pluripotent stem cells (iPSCs) provides a genetically tractable and renewable source of human neural cell populations. We can differentiate these iPSCs into many of the cell types critical for the study of neurotropic viruses, but typically this is performed in monolayer cultures to allow for more control and to generate more homogeneous cell populations, but this methodology lacks the self-organizing properties and interactive dynamics among different cell populations observed during organ development. Recently, more complex structures resembling whole developing organs, named organoids, have been generated from human iPSCs via 3D culturing methods. This emerging new technology has the potential to significantly advance our understanding of infectious diseases and for future therapeutic development. The success of this approach, however, critically depends on how well organoids mimic biological structures, recapitulate human physiology and disease pathology, and incorporate components critical to disease and human host responses. We propose to develop a robust platform for organoid development to model brain development that can be adopted by single labs for basic research, and is amenable to translational studies and drug development and testing. Our Research Center is comprised of three Research Projects, a Scientific Core, and an Administrative Core led by experts in virology, stem cell biology, neural development, and bioengineering. We will focus on ZIKV and WNV, two neurotropic flaviviruses, to develop our organoid platform, which can then be used by the scientific community to investigate other infectious diseases that affect the nervous system. Importantly, ZIKV and WNV are thought to impact the CNS at different stages of development, with ZIKV having been recently implicated as being causal for microcephaly in some pregnant women. This affords us the opportunity to develop an organoid platform with proof-of-principle testing with viruses suspected to have cell type- and stage-specific tropism. Project 1 will focus on technology development to generate more mature organoids and the scaling up of robust assays to perform medium-throughput compound testing. Project 2 will focus on ZIKV infections in early stage organoids and Project 3 will evaluate co-culture organoid systems to model WNV infections in later stage organoids. The projects will be supported by a Scientific Core that will provide cells and on-site training to Projects 2 & 3, as well as optimization of differentiation protocols and bioinformatics analyses. Finally, the Administrative Core will provide logistical support to facilitate collaborations among investigators and to coordinate the timely release of results and resources to the scientific community.

摘要-概述 对影响人类中枢神经系统(CNS)的传染病进行建模，例如 与寨卡病毒(ZIKV)和西尼罗河病毒(WNV)相关的病毒，由于无法访问 相关单元格类型。重新编程人类体细胞，如皮肤成纤维细胞，使其具有诱导多能性 干细胞(IPSCs)为人类神经细胞群体提供了一种遗传上易驯化和可再生的来源。我们 可以将这些ipscs区分成许多对研究嗜神经病毒至关重要的细胞类型，但 通常，这是在单层培养中执行的，以允许更多的控制和生成更同质的 细胞群体，但这种方法缺乏自组织特性和相互作用的动力学 在器官发育过程中观察到不同的细胞群。最近，更复杂的结构类似于 整个发育中的器官，被称为有机体，已经通过3D培养从人类IPSCs中产生 方法：研究方法。这项新兴的新技术有可能极大地提高我们对 传染病和未来的治疗发展。然而，这种方法的成功关键是 这取决于有机化合物模拟生物结构、概括人类生理和疾病的程度。 病理，并包含对疾病和人类宿主反应至关重要的组件。我们建议开发 一个强大的有机体发育平台，用于模拟大脑发育，可供单个实验室采用 基础研究，并服从于转化研究和药物开发和测试。 我们的研究中心由三个研究项目组成，一个科学核心，一个行政管理 核心由病毒学、干细胞生物学、神经发育和生物工程方面的专家领导。我们将重点关注 ZIKV和WNV这两种嗜神经性黄病毒，来开发我们的有机平台，然后可以被 科学界调查影响神经系统的其他传染病。重要的是，ZIKV 和西尼罗河病毒被认为在不同的发展阶段影响中枢神经系统，ZIKV最近 被认为是一些孕妇小头畸形的原因。这为我们提供了一个机会 开发一个有机类平台，对怀疑具有细胞类型的病毒进行原则证明测试-以及 特定阶段的取向。项目1将专注于技术开发，以生产更成熟的有机化合物 以及扩大稳健的测试，以执行中等吞吐量的化合物测试。项目2将重点放在 ZIKV感染早期有机物和项目3将评估共培养有机物系统模型 西尼罗河病毒在后期器官中的感染。这些项目将得到科学核心的支持，该核心将提供 对项目2和3进行细胞和现场培训，以及优化分化方案和 生物信息学分析。最后，行政核心将提供后勤支助，以便利 调查人员之间的合作，并协调及时将结果和资源发布给 科学界。