Grunwell Jocelyn R, Stephenson Susan T, Dallalio Gail A, Diani Badiallo A, Zaworski Celena, Jordan Natalie, Fitzpatrick Anne M
Division of Critical Care Medicine, Children's Healthcare of Atlanta, Arthur M. Blank Hospital, 2220 North Druid Hills Rd NE, Atlanta, GA, 30329, USA.
Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA.
Sci Rep. 2025 May 21;15(1):17703. doi: 10.1038/s41598-025-02078-9.
A challenge that limits our understanding of the underlying pathobiology of pediatric acute respiratory distress syndrome (PARDS) is the lack of a preclinical airway model that can be leveraged for the study of mechanisms and specific molecules for drug testing. We developed a physiologic model system of the small airways for mechanistic application in PARDS using a co-culture of primary human-derived small airway epithelial cells (SAECs) cultured at the air-liquid interface and umbilical vein endothelial cells in a transwell system. The model was validated by exposing the SAECs to a rhinovirus infection, to an inflammatory lung insult using a mixture of cytokines found in ARDS (cytomix), and to airway fluid samples from children with different severity strata of PARDS. We used a combination of transepithelial electrical resistance, immunofluorescence confocal microscopy of tight junctions, targeted gene expression, and cytokine responses to evaluate the model to the aforementioned insults. We then use the model in drug testing and show the reduction in IL-6 expression in conditioned media and STAT3 phosphorylation following co-treatment of SAECs with cytomix and the Janus kinase inhibitor (JAKi) baricitinib. This model enables mechanistic studies of airway pathobiology and may serve as a novel drug testing platform for PARDS.
限制我们对小儿急性呼吸窘迫综合征（PARDS）潜在病理生物学理解的一个挑战是缺乏一种可用于研究药物测试机制和特定分子的临床前气道模型。我们使用在气液界面培养的原代人源小气道上皮细胞（SAECs）与脐静脉内皮细胞在Transwell系统中共培养，开发了一种用于PARDS机制研究的小气道生理模型系统。通过将SAECs暴露于鼻病毒感染、使用急性呼吸窘迫综合征（ARDS）中发现的细胞因子混合物（细胞混合物）进行炎症性肺损伤以及来自不同严重程度分层的PARDS患儿的气道液体样本，对该模型进行了验证。我们结合使用跨上皮电阻、紧密连接的免疫荧光共聚焦显微镜、靶向基因表达和细胞因子反应来评估该模型对上述损伤的反应。然后我们将该模型用于药物测试，并显示在用细胞混合物和Janus激酶抑制剂（JAKi）巴瑞替尼共同处理SAECs后，条件培养基中IL-6表达的降低和STAT3磷酸化的减少。该模型能够对气道病理生物学进行机制研究，并可能作为PARDS的新型药物测试平台。
