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系统疾病，无法治愈。因此，迫切需要新的、安全的、非侵入性的脑部疾病治疗方法。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，进展受到限制。神经组织在经颅磁刺激下的分子/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经颅磁刺激作为治疗神经系统疾病的神经调节工具至关重要。该项目的教育目标是有效地将研究与教育活动结合起来，培养本科生和研究生进行跨学科研究，以培养下一代生物工程师。pi将根据该研究计划开发一个新的垂直整合计划（VIP），题为：利用生物工程方法靶向神经退行性疾病。VIP将以团队为基础，将本科教育和教师研究结合起来。总体教育目标是通过培养学生在良好的技术背景下进行研究，并使他们能够为他们的高级职业获得动手实验技能，从而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，以尽可能地模拟体内条件。这将促进经颅磁刺激对神经退行性疾病的作用和药物筛选试验的研究。该提案的目标是开发一种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，这将允许研究器官发育机制、细胞相互作用、磁场刺激下疾病模型进展和特定微环境下药物治疗。具体而言，建议的努力包括：(1)开发由微室阵列组成的芯片，以便可以有效地制造包括病变神经球（如阿尔茨海默病（AD）神经球）在内的神经球；(2)利用该芯片研究健康神经球和AD神经球在瞬态磁刺激（MS）和药物治疗下的行为。本课题主要创新点如下：(1)利用该微流控芯片，可以快速、廉价地合成具有可调定量组成的大规模神经球，便于对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，可以很容易地制备一系列具有已知淀粉样蛋白-β和/或磷酸化tau浓度的AD模型（AD神经球）；(3)开发该芯片将有助于研究MS和药物治疗对AD模型的影响。该奖项反映了美国国家科学基金会的法定使命，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.