Drug development for tuberous sclerosis complex and other pediatric epileptogenic diseases using neurovascular and cardiac microphysiological models

尽管新冠肺炎主要被认为是一种呼吸道感染，而且大多数死于 疾病都归因于肺衰竭，越来越明显的是，SARS-CoV-2病毒， 直接或间接影响所有主要器官系统，具有令人困惑的可变性程度， 使有效疗法的鉴定复杂化。尤其是中枢神经系统(CNS)和 血管系统似乎都在疾病进展中起着重要作用，而且中枢神经系统的症状与 新冠肺炎患者预后较差。假设中枢神经系统和血管系统相互影响 免疫反应的病理性失调，但人们对它们的反应以及可能的 有助于疾病的发展。目前还没有出现一种能够广泛中和新冠肺炎的治疗剂 疾病进展，这强烈表明任何有效的治疗策略都需要解决 SARS-CoV-2感染不仅对肺部有影响，而且对许多器官系统也有影响，进而 将需要进入中枢神经系统的治疗途径。因此，了解肺和肺之间的相互作用 中枢神经系统对于寻找能够改善新冠肺炎患者预后和减少 住院率和死亡率。该项目将评估SARS-CoV-2在肺部的感染 导致新冠肺炎的器官功能障碍和潜在的中枢神经系统感染，以及 抗病毒和抗炎药物的结合解决了中枢神经系统参与新冠肺炎的问题。这些目标 要求一个生理上相关的体外平台，充分概括系统免疫和细胞因子 与新冠肺炎最严重病例相关的呼吸道上皮感染后的风暴反应 而且可以很容易地在生物安全3级(BSL-3)设施中使用，这是研究高度 传染性呼吸道疾病。该项目将实施一个双器官微生理系统(MPS)模型 将现有的神经血管单位(NVU)/血脑屏障组织芯片用于中枢神经系统组件， 将NVU重新定位为肺部组件的呼吸道芯片，并将两个芯片转换为重力灌注 以便于在BSL-3设施中使用。其目的是1)模拟新冠肺炎感染和先天肺部感染 呼吸道芯片中的响应，2)将NVU和呼吸道芯片耦合以评估呼吸道的响应 芯片到新冠肺炎的感染影响了NVU的功能，需要建立治疗基准 用于药物测试，以及3)筛选FDA批准的药物治疗阴性症状的疗效 NVU/Airway芯片型号。分离的NVU/CNS和呼吸道组织芯片感染的比较 耦合芯片系统的感染将有助于确定每个MPS模型和病毒的传染性 通过血脑屏障进入中枢神经系统的能力。FDA批准的候选药物将接受测试 在两个微生理系统中影响病毒感染、复制和细胞因子产生的能力。

Although COVID-19 is recognized primarily as a respiratory infection and the majority of deaths from the disease are attributed to pulmonary failure, it has become increasingly apparent that the SARS-CoV-2 virus, either directly or indirectly, affects all major organ systems with a confounding degree of variability that complicates the identification of effective therapeutics. In particular, the central nervous system (CNS) and vasculature both seem to play a significant role in disease progression, and CNS symptoms have correlated with poorer outcomes in COVID-19 patients. It is hypothesized that the CNS and vasculature each influence pathological dysregulation of immune response, but very little is known about how they respond and possibly contribute to disease progression. No single therapeutic agent has emerged that broadly neutralizes COVID-19 disease progression, which strongly suggests that any effective treatment strategies will need to address not only effects of SARS-CoV-2 infection in the lungs, but also inflammation in many organ systems, which in turn would require therapeutic access to the CNS. Thus, understanding the interactions between the lungs and the CNS is critical to identifying treatments capable of improving the prognoses of COVID-19 patients and reducing hospitalization rates and mortality. This project will evaluate how SARS-CoV-2 infection in the lungs contributes to both the organ dysfunction in COVID-19 and potential CNS infection, and how well the combination of anti-viral and anti-inflammatory drugs addresses CNS involvement in COVID-19. These goals demand a physiologically relevant in vitro platform that fully recapitulates the systemic immune and cytokine storm responses following infection of airway epithelium associated with the most severe cases of COVID-19 and that can be readily used in the Biosafety Level-3 (BSL-3) facilities required for studies of this highly infectious respiratory disease. This project will implement a two-organ microphysiological system (MPS) model that uses an existing NeuroVascular Unit (NVU)/blood-brain barrier tissue chip for the CNS component, repurposes the NVU as an Airway Chip for the lung component, and converts both chips to gravity perfusion for ease of use in BSL-3 facilities. The aims are to 1) model COVID-19 infection and innate pulmonary response in the Airway Chip, 2) couple the NVU and Airway Chip to evaluate how the response of the Airway Chip to COVID-19 infection affects the function of the NVU, as required to establish therapeutic benchmarks for drug testing, and 3) screen FDA-approved drugs for efficacy in treating negative symptoms in the NVU/Airway Chip model. A comparison of infection of the separate NVU/CNS and Airway tissue chips with infection of the coupled-chip system will help determine the infectability of each MPS model and the viral capacity to cross the blood-brain barrier into the CNS. Candidate FDA-approved drugs will be tested for their ability to affect viral infection, replication, and cytokine production in both microphysiological systems.