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Bioengineering a cortical microtissue model to study human microglia in Alzheimer's disease

生物工程皮质微组织模型来研究阿尔茨海默病中的人类小胶质细胞

基本信息

批准号：
10630949
负责人：
David Allenson Borton
金额：
$ 18.64万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10630949
关键词：
3-Dimensional APP-PS1 Acute Affect Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease therapeutic Amyloidosis Animals Award Behavior Biological Assay Biological Models Biology Biomedical Engineering Brain Characteristics Chimera organism Communities Cues Custom Data Set Development Disease Dissociation Elements Engineering Engraftment Future Gene Expression Generations Genetic Genetic Transcription Genotype Goals Growth Health Human Hybrids Image In Vitro Inflammatory Intervention Knowledge Label Light Link Lipopolysaccharides Mechanics Meta-Analysis Methodology Methods Microglia Mission Modeling Molds Monitor Morphology Mus Mutation Neonatal Nerve Degeneration Neurodegenerative Disorders Neurons Organoids Outcome Pathogenesis Pathogenicity Pathologic Pathology Patients Phenotype Physiological Prevention strategy Property Public Health Publishing Rat Strains Rat Transgene Rattus Research Rest Science Sorting Support System System Techniques Testing Therapeutic Tissue Model Tissues Transgenic Organisms Transplantation United States National Institutes of Health V717F brain cell cell type cost effective design disability falls fighting glial activation improved in vivo induced pluripotent stem cell innovation mutant neural neuroinflammation operation programs reconstitution response scaffold stem cells tau Proteins three dimensional cell culture tool transcriptomics translational study

项目摘要

PROJECT SUMMARY/ABSTRACT Microglia are highly sensitive to the extrinsic cues from their microenvironment in a context-specific manner and rapidly change around diverse cellular states via intrinsic transcriptional programs. The specific microglial state that contributes to Alzheimer’s Disease (AD) pathology is still illy defined, mostly owing to the longstanding critical need for a reliable and robust modeling tool. In light of the substantial difference between human and mouse microglia, the iPSC-derived human microglia (iMG) have emerged as a rigorous platform to investigate neuroinflammation and AD-linked genetics. A recent transition from 2D to 3D culture systems further enhances the utility of iMG in understanding AD pathogenesis, but technical barriers remain. An improvement to achieve better cellular diversity and functionality in a scalable 3D neural setting will make a desired tool that enables precise monitoring and manipulations of iMG inside a stringently controlled brain-like milieu. Our long-term goal is to understand the pathogenic determinants of neuroinflammation to inform crucial future treatment for AD, via stem cell and engineering innovations. To this end, we propose here to introduce an original 3D culture platform to examine the extrinsic and intrinsic elements of microglial activation in health and in AD, based on patient- derived iMG grown in engineered primary rat cortical microtissues. Leveraging validated AD mutant iPSCs and transgenic AD rats, our overall objectives are to 1) confirm the capability of our hybrid model to recapitulate human-specific, AD-related microglia features and signature and 2) to characterize the ‘ground state’ and neuroinflammatory responses of control and AD iMG under a physiological or pathological microenvironment (i.e. wildtype vs. AD transgenic microtissue). Supported by our preliminary studies, our central hypothesis is that the iMG-microtissue growth system supports maturation and operation of human microglia and can be utilized to detect the nuances in microglial behaviors that may contribute to AD pathogenesis. Our specific aims are to test our working hypotheses that 1) iMG populated in cortical microtissues mimic homeostatic human microglia in vivo and can be phenotyped for microglia-intrinsic manifestations contributed by genotypes, and that 2) iMG maintained in AD mutant microtissues are affected by the AD-promoting extrinsic cues and show disease- associated characteristics. Our expected outcomes are to 1) establish a new-generation in vitro human microglia model system that uniquely allows dissecting the intertwined influences from AD-linked genetics and microenvironment on microglial functions and 2) to acquire a distinctive dataset that would offer a framework to pinpoint critical microglia characteristics for AD. We believe an expansion of this research beyond this award period to characterize in depth the microglial findings uncovered here will ultimately bring a valuable opportunity for the development of much-needed intervention and prevention strategies for AD.

项目概要/摘要小胶质细胞以特定的方式对其微环境中的外在线索高度敏感，并且通过内在的转录程序快速改变不同的细胞状态。特定的小胶质细胞状态导致阿尔茨海默病 (AD) 病理学的原因仍然不明确，主要是由于长期存在的关键问题需要可靠且强大的建模工具。鉴于人类和小鼠之间的实质性差异小胶质细胞，iPSC 衍生的人类小胶质细胞 (iMG) 已成为研究的严格平台神经炎症和 AD 相关遗传学。最近从 2D 到 3D 培养系统的转变进一步增强了 IMG 在理解 AD 发病机制方面的效用，但技术障碍仍然存在。要实现的改进可扩展的 3D 神经环境中更好的细胞多样性和功能将成为一种理想的工具，使在严格控制的类脑环境中精确监控和操作 IMG。我们的长期目标的目的是了解神经炎症的致病决定因素，为未来 AD 的关键治疗提供信息干细胞和工程创新。为此，我们在这里建议引入一个原创的3D文化平台根据患者的情况，检查健康和 AD 中小胶质细胞激活的外在和内在因素衍生的 IMG 在工程化的原代大鼠皮质微组织中生长。利用经过验证的 AD 突变 iPSC 和转基因 AD 大鼠，我们的总体目标是 1) 确认我们的混合模型重演的能力人类特异性、AD 相关的小胶质细胞特征和特征，2) 表征“基态”和生理或病理微环境下对照和AD iMG的神经炎症反应（即野生型与 AD 转基因微组织）。在我们的初步研究的支持下，我们的中心假设是 iMG-微组织生长系统支持人类小胶质细胞的成熟和运行，并且可以被利用检测可能导致 AD 发病机制的小胶质细胞行为的细微差别。我们的具体目标是测试我们的工作假设 1) 皮质微组织中的 iMG 模拟稳态的人类小胶质细胞体内并且可以对由基因型贡献的小胶质细胞固有表现进行表型分析，并且 2) iMG AD 突变微组织中维持的细胞受到 AD 促进的外在线索的影响，并表现出疾病- 相关特征。我们的预期结果是 1) 建立新一代体外人类小胶质细胞模型系统独特地允许剖析 AD 相关遗传学的相互交织的影响，微环境对小胶质细胞功能的影响，2) 获取独特的数据集，为查明 AD 的关键小胶质细胞特征。我们相信这项研究的扩展超出了该奖项的范围深入描述此处发现的小胶质细胞发现的时期将最终带来宝贵的机会制定急需的 AD 干预和预防策略。