The Center for Fetal, Cellular, and Molecular Therapy, Cincinnati Children's Hospital Medical Center , Cincinnati, Ohio.
University of Cincinnati School of Medicine , Cincinnati, Ohio.
Am J Physiol Lung Cell Mol Physiol. 2018 Dec 1;315(6):L1028-L1041. doi: 10.1152/ajplung.00148.2018. Epub 2018 Sep 27.
Congenital diaphragmatic hernia (CDH) occurs in ~1:2,000 pregnancies and is associated with substantial morbidity and mortality. Fetal tracheal occlusion (TO) is an emerging therapy that improves lung growth and reduces mortality, although substantial respiratory compromise persists in survivors. In this study, we used tracheal fluid in a fetal sheep model of CDH with TO for proteomic analysis with subsequent validation of findings in sheep lung tissue. We found that the proteomic profiles of CDH tracheal fluid was most similar to control lung and CDH/TO lung most similar to TO lung. Among 118 proteins altered in CDH, only 11 were reciprocally regulated in CDH/TO. The most significantly altered pathways and processes were cell proliferation, phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin signaling, inflammation, and microtubule dynamics. CDH suppressed and TO promoted cell proliferation and AKT-related signaling cascades. By Western blot analysis and immunohistochemistry, epithelial PCNA and phosphorylated AKT were decreased in CDH and increased in TO and CDH/TO lungs. The Wnt target Axin2 was decreased threefold in CDH lung compared with control without a significant increase in CDH/TO lung. Cilia-related pathways were among the most dysregulated with CDH lung having a nearly twofold increase in acetylated α-tubulin and a relative increase in the number of ciliated cells. While TO improves lung growth and patient survival in CDH, the procedure substantially alters many processes important in lung development and cell differentiation. Further elucidation of these changes will be critical to improving lung health in infants with CDH treated with TO.
先天性膈疝 (CDH) 在约 1:2000 的妊娠中发生，与大量发病率和死亡率相关。胎儿气管阻塞 (TO) 是一种新兴的治疗方法，可改善肺生长并降低死亡率，但幸存者仍存在严重的呼吸功能障碍。在这项研究中，我们使用了 TO 治疗的 CDH 胎儿羊模型中的气管液进行蛋白质组学分析，并随后在绵羊肺组织中验证了研究结果。我们发现，CDH 气管液的蛋白质组学特征与对照肺最相似，而 CDH/TO 肺与 TO 肺最相似。在 118 种改变的 CDH 蛋白中，只有 11 种在 CDH/TO 中呈反向调节。改变最显著的途径和过程是细胞增殖、磷脂酰肌醇 3-激酶/AKT/雷帕霉素哺乳动物靶蛋白信号转导、炎症和微管动力学。CDH 抑制而 TO 促进细胞增殖和 AKT 相关信号级联。通过 Western blot 分析和免疫组织化学分析，上皮细胞 PCNA 和磷酸化 AKT 在 CDH 中减少，而在 TO 和 CDH/TO 肺中增加。与对照相比，CDH 肺中的 Wnt 靶标 Axin2 减少了三倍，而 CDH/TO 肺中没有明显增加。与 CDH 肺相比，与 cilia 相关的途径发生了最大程度的失调，CDH 肺中的乙酰化 α-微管蛋白增加了近两倍，纤毛细胞的数量也相对增加。尽管 TO 改善了 CDH 患者的肺生长和患者生存率，但该手术会极大地改变许多对肺发育和细胞分化很重要的过程。进一步阐明这些变化对于提高接受 TO 治疗的 CDH 婴儿的肺健康至关重要。
