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

尽管COVID-19主要被认为是一种呼吸道感染， 疾病归因于肺衰竭，越来越明显的是SARS-CoV-2病毒， 直接或间接地影响所有主要器官系统，具有混杂程度的变异性， 使有效治疗剂的鉴定复杂化。特别是中枢神经系统（CNS）和 血管系统似乎在疾病进展中起重要作用，CNS症状与疾病进展相关。 在COVID-19患者中的结果较差。假设中枢神经系统和血管系统各自影响 免疫反应的病理性失调，但对它们如何反应知之甚少， 有助于疾病进展。目前还没有出现一种能广泛中和COVID-19的治疗药物 疾病进展，这强烈表明，任何有效的治疗策略都需要解决不 只有SARS-CoV-2感染在肺部的影响，而且在许多器官系统中也有炎症， 需要进入中枢神经系统进行治疗因此，了解肺和肺之间的相互作用， CNS对于确定能够改善COVID-19患者病情并减少 住院率和死亡率。该项目将评估SARS-CoV-2如何在肺部感染 导致COVID-19中的器官功能障碍和潜在的CNS感染， 抗病毒和抗炎药物的组合解决了CNS参与COVID-19的问题。这些目标 需要一种生理学相关的体外平台，其完全概括全身免疫和细胞因子 与最严重的COVID-19病例相关的气道上皮感染后的风暴反应 并且可以很容易地用于生物安全3级（BSL-3）设施，用于研究这种高度生物安全的生物。 传染性呼吸道疾病本计画将实作一个双器官微生理系统（MPS）模型 该技术使用现有的神经血管单元（NVU）/血脑屏障组织芯片作为中枢神经系统组件， 将NVU重新用作肺部组件的气道芯片，并将两种芯片转换为重力灌注 以便于在BSL-3设施中使用。目的是1）模拟COVID-19感染和先天性肺 2）将NVU与气道芯片耦合以评估气道芯片的响应如何与NVU的响应相关联， 根据建立治疗基准的要求，COVID-19感染的芯片会影响NVU的功能 进行药物测试，3）筛选FDA批准的药物，以确定其治疗阴性症状的有效性。 NVU/气道芯片型号。比较单独的NVU/CNS和气道组织芯片的感染， 耦合芯片系统的感染将有助于确定每个MPS模型和病毒的感染性， 穿过血脑屏障进入中枢神经系统的能力。FDA批准的候选药物将被测试其 在两种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.