Novel tools for in vitro electrophysiology and neurotrauma modeling

在美国，创伤性脑损伤本身仍然是导致死亡和残疾的一个重要原因。 而且是其他神经退行性疾病的重要风险因素。然而，目前有 目前还没有批准的治疗脑损伤的方法，其长期后果很难预测。30多个专业 第三阶段的试验失败了，没有一次成功，因此通用疗法的发现似乎越来越多 不太可能。NINDS和其他联邦机构已经承诺向大型企业提供数千万美元， 观察性的、人体的脑损伤研究。这些研究是对脑外伤患者进行基因分型和深度表型分析 以个人化治疗为目标。这些努力已经揭示了令人着迷的相关性 基因分型和脑损伤转归。然而，出于伦理原因，人类的基因不能被开启和关闭。 因此，需要新的工具来从检测相关性转向测试假说。这 在使用人类体外模型的其他疾病中，已经解决了挑战。人类产生的神经元 使用干细胞技术的患者保留了患者的遗传身份。此外，基因变异也可以 在这些单元格里一次换一个。因此，关于基因在疾病中的作用的假设可以 可以在人体的体外模型中进行测试，但前提是这种疾病的病理可以在体外复制。再生产 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一种机械侮辱。 这项提案的目标是为体外模拟神经创伤提供新的工具，这些工具可以利用 人类体外培养中令人兴奋的最新发展。靶向药物发现在中国是困难的 神经损伤，因为分子机制很复杂。因此，表型药物的发现 更可取，但只有当它解决临床相关的表型时，它才能成功。在体外，电场 录音之所以吸引人，是因为它们类似于常用的脑电图法。 对脑外伤患者进行评估。这项工作将有助于第一个能够获得场的多电极阵列(MEA 来自高通量的体外模型的录音。脑器官复制了疾病的某些方面，但不能 在2D培养中复制。然而，电场记录很难从大脑中获取 有机化合物，因为传统的多电极阵列是为贴壁培养而设计的，而大脑 有机化合物需要超低的附着条件。因此，新的亚毫米级结构是 提议将有机类物质封闭在一组电极中，而不是附着它，因此从长远来看 可以对电活动和连接性进行测量。这些3D Mea将为以下方面提供新的见解 除神经创伤外，还有许多神经疾病。目前，没有可用的工具可以应用 对有机体培养的生物忠实、机械的侮辱。拟议的工作将开发这样一个工具。在……里面 结合起来，这些新工具将在个性化治疗、探索 疾病机制，并提供针对患者的风险评估。

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.