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

在美国，18岁以上的美国人中约有五分之一患有可诊断的神经系统疾病，但没有治愈的见解。因此，迫切需要用于治疗脑疾病的新的、安全的、非侵入性的方法。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些成功。然而，由于对磁场与神经组织的相互作用了解不足，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，研究瞬态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程序和实现TMS作为治疗神经系统疾病的神经调节工具的使用至关重要。该项目的教育目标是有效地将研究与教育活动相结合，培养跨学科研究的本科生和研究生，以培养下一代生物工程师。PI将根据该研究提案开发一个新的垂直整合计划（VIP），该研究提案题为：使用生物工程方法靶向神经退行性疾病。VIP将在以团队为基础的背景下将本科教育和教师研究结合起来。 总体教育目标是通过培训学生以良好的技术背景开展研究，并让他们获得实践实验室技能，以帮助下一代劳动力的发展。长期目标是设计一个自动化的技术平台来合成各种体外中枢神经系统疾病模型，以尽可能接近地模拟体内条件。该项目的目标是开发一种基于芯片的微流体平台，该平台有助于快速形成中枢神经系统的三维体外细胞培养模型，这将允许研究器官发育，细胞相互作用，在磁场刺激和药物治疗下的疾病模型进展在定义的微环境中。 具体而言，提出的努力包括（1）开发由微室阵列组成的芯片，使得可以以有效的方式制造包括患病神经球如阿尔茨海默病（AD）神经球的神经球;和（2）使用该芯片研究健康神经球和AD神经球在瞬时磁刺激（MS）和药物治疗下的行为。本研究的主要创新点在于：（1）利用这种微流控芯片，可以快速、廉价地合成大规模的、成分可调的、定量的神经球，为不同类型神经球的研究提供了便利;（2）在该芯片的上游使用浓度梯度发生器，一系列AD模型可以容易地制造具有已知浓度的淀粉样蛋白-β和/或磷酸化-tau的AD神经球;以及（3）因此，开发这种芯片将有助于研究MS和药物治疗对AD模型的影响。该奖项反映了NSF的法定使命并通过使用基金会的知识价值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.