Studies of neurospheres and diseased neurospheres on chip under magnetic field stimulation and drug treatment

在美国，大约五分之一的18岁以上的美国人患有可诊断的神经疾病，但没有治愈的洞察力。因此，迫切需要新的、安全的、非侵入性的治疗脑部疾病的方法。包括重复经颅磁刺激(TMS)和经颅直流电刺激在内的非侵入性技术已经取得了一定的成功。然而，由于对磁场与神经组织的相互作用知之甚少，这方面的研究进展有限。TMS作用下神经组织的分子/细胞机制尚不清楚。因此，研究瞬变磁场对成人神经发生、细胞分化和神经组织(神经球)可塑性的影响对于开发新的治疗方法和实现TMS作为神经调节工具用于神经疾病的治疗是至关重要的。该项目的教育目标是有效地将研究与教育活动相结合，并在跨学科研究方面培养本科生和研究生，以培养下一代生物工程师。PIS将基于这项名为：使用生物工程方法瞄准神经退行性疾病的研究建议，开发一个新的垂直整合计划(VIP)。VIP将在基于团队的背景下将本科教育和教师研究统一起来。教育的总体目标是通过培训学生进行具有良好技术背景的研究，并使他们能够为自己的高级职业获得动手实验室技能，从而帮助下一代劳动力发展。长期目标是设计一个自动化的技术平台，以合成各种体外中枢神经系统疾病模型，以尽可能接近体内的条件。这将有助于TMS效应的研究和神经退行性疾病的药物筛选试验。该建议的目标是开发一个基于芯片的微流体平台，促进中枢神经系统三维体外细胞培养模型的快速形成，这将允许在DefiNed微环境中研究器官发育、细胞相互作用、磁场刺激下的疾病模型进展和药物治疗的机制。具体地说，建议的工作包括(1)开发一种由微腔阵列组成的芯片，以便能够以高效的方式制造包括阿尔茨海默病(AD)神经球等患病神经球的神经球；以及(2)使用该芯片研究健康神经球和AD神经球在瞬变磁刺激(MS)和药物治疗下的行为。本项目的主要创新点可概括为：(1)利用这种微流控芯片，可以快速、廉价地合成组成可调的大规模神经球，便于不同类型神经球的研究；(2)利用该芯片上游的浓度梯度发生器，可以容易地制造出一系列已知淀粉样蛋白-β和/或磷酸化-tau浓度的AD模型(AD神经球)；(3)开发这种芯片将有助于研究多发性硬化症和药物治疗对AD模型的影响。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

In the United States, about one in five Americans above the age of 18 suffer from diagnosable neurological disorders with no cure insight. As such, new, safe, non-invasive methods for the treatment of brain disorders are critically needed. Non-invasive techniques including repetitive transcranial magnetic stimulation (TMS) and transcranial direct current stimulation have had some success. However, progress has been limited due to poor understanding of the interactions of magnetic fields with nervous tissue. The molecular/cellular mechanisms of nervous tissue under TMS are still lacking. Hence, investigation of effects of transient magnetic fields on adult neurogenesis, cell differentiation and plasticity of nervous tissue (neurospheres) is essential in developing new treatment procedures and achieving the use of TMS as a neuromodulation tool for treating neurological disorders. The educational goal of this project is to effectively integrate research with educational activities and to train both undergraduate and graduate students in interdisciplinary studies to produce next-generation bioengineers. The PIs will develop a new Vertically Integrated Program (VIP) based on this research proposal entitled: Targeting Neurodegenerative Diseases Using Bioengineering Approaches. The VIP will unite undergraduate education and faculty research in a team-based context. The overall educational goal is to help next-generation workforce development by training students to carry out research with sound technical background and allowing them to gain hands-on laboratory skills for their advanced careers. The long-term goal is to design an automatic technical platform to synthesize a variety of in vitro central nervous system disease models to mimic in vivo conditions as closely as possible. This will facilitate the studies of TMS effects and drug screening assays for neurodegenerative disorders.The goal of this proposal is to develop a chip-based microfluidics platform that facilitates the rapid formation of three-dimensional in vitro cell culture models of the central nervous system, which will permit the investigation of mechanisms of organ development, cellular interactions, disease model progression under magnetic field stimulation and drug treatments within defined microenvironments. Specifically, the proposed efforts include (1) the development of a chip consisting of microchamber arrays so that neurospheres including diseased neurospheres such as Alzheimer’s disease (AD) neurospheres can be fabricated in an efficient manner; and (2) the studies of the behavior of healthy neurospheres and AD neurospheres under transient magnetic stimulation (MS) and drug treatment using this chip. Major innovations of this proposed project can be summarized as the following: (1) Using this type of microfluidic chip, large-scale neurospheres with tunable and quantitative compositions can be synthesized rapidly and inexpensively, facilitating studies of different types of neurospheres; (2) Using a concentration gradient generator at the upper stream of this chip, a series of AD models (AD neurospheres) with known concentrations of amyloid-β and/or phosphorylated-tau can be readily fabricated; and (3) developing this chip will thus facilitate studies of the effects of both MS and drug treatment on AD models.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.