Marsico Lung Institute/Cystic Fibrosis Research Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States.
Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States.
Am J Physiol Lung Cell Mol Physiol. 2024 Mar 1;326(3):L226-L238. doi: 10.1152/ajplung.00421.2022. Epub 2023 Dec 27.
Cell therapy is a potential treatment for cystic fibrosis (CF). However, cell engraftment into the airway epithelium is challenging. Here, we model cell engraftment in vitro using the air-liquid interface (ALI) culture system by injuring well-differentiated CF ALI cultures and delivering non-CF cells at the time of peak injury. Engraftment efficiency was quantified by measuring chimerism by droplet digital PCR and functional ion transport in Ussing chambers. Using this model, we found that human bronchial epithelial cells (HBECs) engraft more efficiently when they are cultured by conditionally reprogrammed cell (CRC) culture methods. Cell engraftment into the airway epithelium requires airway injury, but the extent of injury needed is unknown. We compared three injury models and determined that severe injury with partial epithelial denudation facilitates long-term cell engraftment and functional CFTR recovery up to 20% of wildtype function. The airway epithelium promptly regenerates in response to injury, creating competition for space and posing a barrier to effective engraftment. We examined competition dynamics by time-lapse confocal imaging and found that delivered cells accelerate airway regeneration by incorporating into the epithelium. Irradiating the repairing epithelium granted engrafting cells a competitive advantage by diminishing resident stem cell proliferation. Intentionally, causing severe injury to the lungs of people with CF would be dangerous. However, naturally occurring events like viral infection can induce similar epithelial damage with patches of denuded epithelium. We found that viral preconditioning promoted effective engraftment of cells primed for viral resistance. Cell therapy is a potential treatment for cystic fibrosis (CF). Here, we model cell engraftment by injuring CF air-liquid interface cultures and delivering non-CF cells. Successful engraftment required severe epithelial injury. Intentionally injuring the lungs to this extent would be dangerous. However, naturally occurring events like viral infection induce similar epithelial damage. We found that viral preconditioning promoted the engraftment of cells primed for viral resistance leading to CFTR functional recovery to 20% of the wildtype.
细胞治疗是囊性纤维化 (CF) 的一种潜在治疗方法。然而，将细胞植入气道上皮是具有挑战性的。在这里，我们通过在气道上皮的气液界面 (ALI) 培养系统中损伤分化良好的 CF ALI 培养物并在损伤高峰时输送非 CF 细胞来模拟细胞植入。通过液滴数字 PCR 测量嵌合体和在 Ussing 室中测量功能离子转运来量化植入效率。使用这种模型，我们发现当用人支气管上皮细胞 (HBEC) 条件重编程细胞 (CRC) 培养方法培养时，细胞的植入效率更高。细胞植入气道上皮需要气道损伤，但所需的损伤程度尚不清楚。我们比较了三种损伤模型，发现严重损伤伴部分上皮脱落有利于长期细胞植入和功能 CFTR 恢复，最高可达野生型功能的 20%。气道上皮迅速对损伤做出反应而再生，这为有效的植入创造了空间竞争并构成了障碍。我们通过延时共聚焦成像检查了竞争动力学，发现输送的细胞通过整合到上皮中来加速气道再生。照射修复的上皮通过减少常驻干细胞增殖赋予植入细胞竞争优势。故意对 CF 患者的肺部造成严重损伤是危险的。然而，像病毒感染这样的自然事件可以诱导类似的上皮损伤，形成裸露的上皮斑块。我们发现病毒预处理促进了对病毒有抗性的细胞的有效植入。细胞治疗是囊性纤维化 (CF) 的一种潜在治疗方法。在这里，我们通过损伤 CF 气液界面培养物并输送非 CF 细胞来模拟细胞植入。成功的植入需要严重的上皮损伤。故意将肺部损伤到这种程度是危险的。然而，像病毒感染这样的自然事件会诱导类似的上皮损伤。我们发现病毒预处理促进了对病毒有抗性的细胞的植入，使 CFTR 功能恢复到野生型的 20%。
