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）有关相关细胞类型。将人类体细胞（例如皮肤成纤维细胞）重新编程为诱导的多能干细胞（IPSC）提供了人类神经细胞群体的一般易和可再生能源。我们可以将这些IPSC区分为许多对神经性病毒研究至关重要的细胞类型，但通常，这是在单层培养物中进行的，以允许更多的控制并产生更多均匀细胞种群，但是这种方法缺乏自组织的特性和交互动力在器官发育过程中观察到的不同细胞群体。最近，更复杂的结构类似于整个发育中的器官，名为类器官，是通过3D培养从人IPSC产生的方法。这项新兴的新技术有可能大大提高我们对传染病和未来的治疗发展。然而，这种方法的成功批评取决于类器官的模拟生物结构的效果如何，概括了人类的生理和疾病病理学，并结合对疾病和人类宿主反应至关重要的成分。我们建议发展一个强大的器官开发平台，用于建模大脑开发，单个实验室可以采用基础研究，并且可以接受翻译研究，药物开发和测试。我们的研究中心由三个研究项目组成，一个科学核心和一个行政由病毒学，干细胞生物学，神经发育和生物工程专家领导的核心。我们将重点关注 ZIKV和WNV，两个神经性黄素毒素，以开发我们的类器官平台，然后可以由该平台使用科学界调查影响神经系统的其他传染病。重要的是，zikv WNV被认为会在开发的不同阶段影响中枢神经系统，而Zikv最近已经在某些孕妇中被实施为小头畸形的因果。这为我们提供了机会通过涉嫌具有细胞类型和细胞类型的病毒来开发带有原理测试的类器官平台特定于阶段的向向主义。项目1将重点介绍技术开发以产生更多成熟的类器官并缩放强大的阿萨斯以执行中等通量化合物测试。项目2将重点关注早期类型器官和项目3中的ZIKV感染将评估共培养器官系统以建模 WNV感染后期的类器官。这些项目将得到科学核心的支持，该核心将提供细胞和现场培训对项目2和3，以及分化方案的优化和生物信息学分析。最后，行政核心将提供后勤支持以促进调查人员之间的合作，并协调及时发布结果和资源科学界。