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

基本信息

批准号：
9913453
负责人：
Guo-li Ming
金额：
$ 151.19万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
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/9913453
关键词：
3-Dimensional 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 Viral Pathogenesis 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

项目摘要

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 – 概述影响人类中枢神经系统 (CNS) 的传染病建模，例如与寨卡病毒（ZIKV）和西尼罗河病毒（WNV）相关的病毒，由于难以接近而一直具有挑战性。将人类体细胞（例如皮肤成纤维细胞）重新编程为诱导多能细胞。干细胞（iPSC）为人类神经细胞群提供了遗传上易于处理且可再生的来源。可以将这些 iPSC 分化为许多对嗜神经病毒研究至关重要的细胞类型，但是通常，这是在单层培养物中进行，以允许更多的控制并产生更均质的细胞群，但这种方法缺乏自组织特性和细胞群之间的相互作用动态最近，在器官发育过程中观察到了不同的细胞群，结构更加复杂。完整的发育器官，称为类器官，是通过 3D 培养从人类 iPSC 中生成的这种新兴的新技术有可能显着增进我们对方法的理解。然而，这种方法的成功对于传染病和未来的治疗发展至关重要。取决于类器官模仿生物结构、重现人类生理和疾病的程度我们建议开发对疾病和人类宿主反应至关重要的成分。一个强大的类器官开发平台，用于模拟大脑发育，可由单个实验室采用基础研究，并适合转化研究以及药物开发和测试。我们的研究中心由三个研究项目、一个科学核心项目和一个行政项目组成核心由病毒学、干细胞生物学、神经发育和生物工程专家领导。 ZIKV 和 WNV，两种嗜神经黄病毒，用于开发我们的类器官平台，然后可供科学界正在研究影响神经系统的其他传染病。和 WNV 被认为会在不同的发展阶段影响中枢神经系统，而 ZIKV 最近已被认为被认为是某些孕妇小头畸形的原因，这为我们提供了机会。开发一个类器官平台，对疑似具有细胞类型和细胞类型的病毒进行原理验证测试项目 1 将侧重于技术开发以生成更成熟的类器官。项目 2 将重点关注扩大稳健检测以进行中等通量化合物测试。早期类器官中的 ZIKV 感染和项目 3 将评估共培养类器官系统以建模后期类器官中的西尼罗河病毒感染将得到科学核心的支持，该核心将提供细胞和项目 2 和 3 的现场培训，以及分化方案的优化和最后，行政核心将提供后勤支持以促进生物信息学分析。调查人员之间的合作，并协调及时向研究人员发布结果和资源科学界。