Novel tools for in vitro electrophysiology and neurotrauma modeling

用于体外电生理学和神经创伤建模的新工具

基本信息

批准号：
10573222
负责人：
John D Finan
金额：
$ 60.83万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-17 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10573222
关键词：
3-Dimensional Accelerometer Address Adherence Adherent Culture Biological Sciences Biomedical Engineering Blood Brain Caring Cause of Death Cell Culture Techniques Cells Collaborations Complex Data Development Disease Electrodes Electroencephalography Electrophysiology (science)Engineering Ethical Issues Ethics Etiology Experimental Genetics Genes Genetic Identity Genome Genotype Goals Helmet Human In Vitro Measures Mechanics Mission Modeling Molecular National Institute of Neurological Disorders and Stroke Nerve Degeneration Nervous System Trauma Neurons Organoids Outcome Output Pathology Patients Phenotype Population Proteomics Public Health Research Rest Risk Risk Assessment Risk Factors Role Secure Skin Stretching Structure TBI Patients TBI treatment Testing Time Trauma Traumatic Brain Injury United States Work behavior test candidate identification clinically relevant design disability drug discovery electric field experimental study fascinate flexibility genetic variant in vitro Model insight millimeter multi-electrode arrays multidisciplinary nervous system disorder neural novel personalized medicine phase III trial physical science precision medicine response serial imaging stem cell technology stem cells success synergism three dimensional cell culture tool

项目摘要

Traumatic brain injury (TBI) remains a significant cause of death and disability in its own right in the United States and is an important risk factor for other neurodegenerative conditions. However, there are currently no approved therapies for TBI and its long term consequences are difficult to predict. More than 30 major phase III trials have failed without a single success so discovery of a universal therapy seems increasingly unlikely. NINDS and other federal agencies have committed tens of millions of dollars to large, observational, human studies of TBI. These studies are genotyping and deeply phenotyping TBI patients with the goal of personalizing therapy. These efforts have already revealed fascinating correlations between genotype and TBI outcome. However, genes cannot be switched on an off in humans for ethical reasons. Therefore, new tools are necessary to move from detecting correlations to testing hypotheses. This challenge has been addressed in other diseases using human, in vitro models. Human neurons generated from patients using stem cell technology retain the genetic identity of the patient. Also, genetic variants can be changed one at a time in these cells. Therefore, hypotheses about the role of genotype in disease can be tested in human, in vitro models but only if the disease pathology can reproduced in vitro. Reproducing neurotrauma pathology in vitro requires special tools because it depends intrinsically on a mechanical insult. The goal of this proposal is to provide new tools for modeling neurotrauma in vitro that can take advantage of exciting recent developments in human, in vitro cultures. Target-driven drug discovery is difficult in neurotrauma because the molecular mechanisms are complex. Phenotypic drug discovery is therefore preferable but it can succeed only if it addresses a clinically relevant phenotype. In vitro, electrical field recordings are attractive because they are analogous to electroencephalography, which is commonly used to assess TBI patients. This work will contribute the first, multi-electrode array (MEA) that can acquire field recordings from a high throughput, in vitro model. Brain organoids reproduce aspects of disease that cannot be reproduced in 2D cultures. However, electrical field recordings are difficult to acquire from brain organoids because conventional, multi-electrode arrays are designed for adherent cultures while brain organoids require ultra-low adherence conditions. Therefore, novel, sub-millimeter scale structures are proposed that will enclose an organoid inside an array of electrodes without adhering to it so that long term measures of electrical activity and connectivity can be made. These 3D MEAs will contribute new insights to many neurological disorders beside neurotrauma. Currently, there are no tools available that can apply a biofidelic, mechanical insult to an organoid culture. The proposed work will develop such a tool. In combination, these new tools will open new horizons in the field around personalizing therapy, probing disease mechanism and offering patient-specific risk assessment.

创伤性脑损伤 (TBI) 本身仍然是美国死亡和残疾的一个重要原因是其他神经退行性疾病的重要危险因素。然而，目前有目前还没有批准的治疗 TBI 的疗法，其长期后果很难预测。 30多个专业 III 期试验失败了，没有一次成功，因此似乎越来越多地发现通用疗法不太可能。 NINDS 和其他联邦机构已承诺向大型、 TBI 的观察性人体研究。这些研究对 TBI 患者进行基因分型和深度表型分析以实现个性化治疗的目标。这些努力已经揭示了之间令人着迷的相关性基因型和 TBI 结果。然而，出于伦理原因，人类的基因不能被打开或关闭。因此，需要新的工具从检测相关性转向检验假设。这已经使用人体体外模型解决了其他疾病中的挑战。人类神经元产生使用干细胞技术从患者身上保留患者的遗传身份。此外，基因变异也可以在这些单元格中一次更改一个。因此，关于基因型在疾病中的作用的假设可以可以在人体体外模型中进行测试，但前提是疾病病理学可以在体外重现。繁殖体外神经创伤病理学需要特殊的工具，因为它本质上依赖于机械损伤。该提案的目标是提供体外神经创伤建模的新工具，可以利用人类体外培养中令人兴奋的最新进展。靶点驱动的药物发现是困难的神经损伤，因为其分子机制很复杂。因此，表型药物发现更好，但只有解决临床相关表型才能成功。体外，电场录音很有吸引力，因为它们类似于常用的脑电图评估 TBI 患者。这项工作将贡献第一个可以获取场的多电极阵列（MEA）来自高通量体外模型的记录。大脑类器官能够再现疾病的某些方面，而这些方面是无法再现的。可以在二维文化中复制。然而，很难从大脑中获取电场记录类器官，因为传统的多电极阵列是为贴壁培养而设计的，而大脑类器官需要超低依从性条件。因此，新颖的亚毫米级结构提议将类器官封装在电极阵列内而不粘附它，以便长期可以进行电活动和连接性的测量。这些 3D MEA 将为除了神经外伤之外的许多神经系统疾病。目前，没有可用的工具可以应用对类器官培养物的生物忠实的机械损伤。拟议的工作将开发这样一个工具。在结合起来，这些新工具将在个性化治疗、探索领域开辟新视野疾病机制并提供针对患者的风险评估。