EAGER: AIR-NCS Novel epigenetic assay to probe neural stem cell fate in neurodegenerative environment

概述：这项热切的提议是对加速神经科学和认知科学综合研究(AIR-NCS)倡议的响应。这项提议寻求开发新的工具，可以用来研究神经退行性环境如何导致神经干细胞的表观遗传变化，从而影响分化模式，并最终影响大脑如何适应和响应神经退行性环境。该提案具体回应了主题领域“适应不断变化的环境”。这项工作将生物工程(开发一种新的表观遗传学分析)与神经科学和发育生物学(研究神经退化环境如何影响神经干细胞的命运)结合起来，应该会影响生物医学工程计划感兴趣的神经再生领域。这项拟议的工作适合于渴望机制，因为它是探索性的，基于一个新的、未经证实的假设，但如果成功，可能会在工程学和神经科学之间的界面上带来新的变革性发现。智力优势：神经干细胞分化为不同神经元和神经胶质表型的过程需要与基因沉默和/或激活相关的程序之间复杂的相互作用。某些基因启动子区域的DNA甲基化是动态调节基因表达的一种机制。假设某些神经退行性环境诱导神经干细胞的表观遗传学改变，导致细胞增殖和/或分化的改变，这有助于疾病的表型，并限制干细胞治疗的神经再生潜力。具体地说，假设与高氧化应激和β-淀粉样蛋白的存在相关的模型神经退行性变(阿尔茨海默病样)环境将导致与SMAD或JAK/STAT通路相关的启动子的DNA甲基化改变，导致神经干细胞命运的改变，例如从神经发生转向反应性星形胶质细胞的形成。为了验证这一新假说，建议(1)开发一种新的工具来快速、廉价地检测培养细胞对环境刺激的表观遗传学变化；(2)使用新工具来检测神经干细胞对模型神经退行性(阿尔茨海默氏症)环境的表观遗传学变化；以及(3)评估表观遗传学变化与神经干细胞分化的变化有何相似之处，以及这如何影响干细胞治疗的神经再生潜力。虽然最终目标是解决所有这些目标，并要求该机构提供为期一年的资金，但计划的工作将集中在开发用于快速探测细胞表观遗传学变化的新工具，然后解决在神经干细胞中使用该工具的可行性。这项工作将有助于理解不同环境如何影响干细胞分化，以及表观遗传学在细胞对环境反应中的作用。广泛影响：拟议的工作最终将有助于开发基于干细胞的神经再生策略。一种快速和廉价地检测神经干细胞和干细胞分化的表观遗传学变化的新工具也可以用于检测与癌症或其他疾病相关的表观遗传学变化。表观遗传学实验以热力学原理为基础，并将被用作密歇根大学化学工程课程的核心课程以及国际生物工程研究所教授的生物工程实验室的例子。一名本科生研究人员和一名研究生将参与该项目，并在跨学科环境中接受培训。

Overview:This EAGER proposal responds to the Accelerating Integrative Research in Neuroscience and Cognitive Science (AIR-NCS) initiative. This proposal seeks to develop novel tools that can be used to examine how neurodegenerative environments lead to epigenetic changes in neural stem cells, which then impact patterns of differentiation and ultimately affect how the brain adapts and responds to the neurodegenerative environment. This proposal responds specifically to the thematic area "adaptation to changing environments". The work combined bioengineering (development of a novel epigenetic assay) with neuroscience and developmental biology (examining how the neurodegenerative environment impacts neural stem cell fate), and should impact areas of neuroregeneration of interest to the Biomedical Engineering Program. The proposed work is appropriate for an EAGER mechanism as it is exploratory, based on a new, unproven hypothesis, but if successful, could lead to new transformational discoveries at the interface between engineering and neuroscience.Intellectual Merit:The process of neural stem cell differentiation into different neuronal and glial phenotypes requires the complex interplay between programs associated with gene silencing and or activation. DNA methylation of promoter regions of certain genes is one mechanism by which gene expression is dynamically regulated. It is hypothesized that certain neurodegenerative environments induce epigenetic changes in neural stem cells, leading to changes in cell proliferation and/or differentiation, which contributes to the disease phenotype and limit the neuroregenerative potential of stem cell therapies. Specifically, it is hypothesized that a model neurodegenerative (Alzheimer's like) environment associated with high oxidative stress and the presence of beta-amyloid will lead to changes in DNA methylation of promoters associated with either the SMAD or JAK/STAT pathways, leading to changes in neural stem cell fate such as a shift away from neurogenesis and towards the formation of reactive astrocytes. To test this novel hypothesis, it is proposed (1) to develop a novel tool to rapidly and inexpensively probe epigenetic changes in cultured cells in response to an environmental stimulus; (2) to use the new tool to examine epigenetic changes in neural stem cells in response to a model neurodegenerative (Alzheimer's like) environment; and (3) to assess how epigenetic changes are liked to changes in neural stem cell differentiation, and how that impacts the neuroregenerative potential of stem cell therapies. While the ultimate goal is to address all of these goals, with funding for one year requested from this EAGER, the work planned will be focused on the development of the novel tool for rapidly probing epigenetic changes in cells and then address the feasibility of using the tool in neural stem cells. This work will contribute to the understanding of how different environments affect stem cell differentiation, and the role of epigenetics in cellular response to the environment.Broader Impacts:The proposed work will eventually assist in the development of neuroregenerative strategies based on stem cells. A novel tool for rapidly and inexpensively examining epigenetic changes in neural stem cells and stem cell differentiation could also be used to examine epigenetic changes associated with cancer or other diseases. The epigenetic assay is based on thermodynamic principles, and will be used as an example in a core course in the chemical engineering curriculum at UMBC as well as the bioengineering laboratory taught by the PI. One undergraduate researcher and one graduate student will work on the project and receive training in an interdisciplinary environment.