Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
School of Respiratory Therapy, College of Medicine, Taipei Medical University, 250 Wuxing Street, Taipei, 11031, Taiwan.
Mol Med. 2023 Nov 23;29(1):159. doi: 10.1186/s10020-023-00760-0.
Delay in type II alveolar epithelial cell (AECII) regeneration has been linked to higher mortality in patients with acute respiratory distress syndrome (ARDS). However, the interaction between Doublecortin-like kinase 1 (DCLK1) and the Hippo signaling pathway in ARDS-associated AECII differentiation remains unclear. Therefore, the objective of this study was to understand the role of the DCLK1/Hippo pathway in mediating AECII differentiation in ARDS.
AECII MLE-12 cells were exposed to 0, 0.1, or 1 μg/mL of lipopolysaccharide (LPS) for 6 and 12 h. In the mouse model, C57BL/6JNarl mice were intratracheally (i.t.) injected with 0 (control) or 5 mg/kg LPS and were euthanized for lung collection on days 3 and 7.
We found that LPS induced AECII markers of differentiation by reducing surfactant protein C (SPC) and p53 while increasing T1α (podoplanin) and E-cadherin at 12 h. Concurrently, nuclear YAP dynamic regulation and increased TAZ levels were observed in LPS-exposed AECII within 12 h. Inhibition of YAP consistently decreased cell levels of SPC, claudin 4 (CLDN-4), galectin 3 (LGALS-3), and p53 while increasing transepithelial electrical resistance (TEER) at 6 h. Furthermore, DCLK1 expression was reduced in isolated human AECII of ARDS, consistent with the results in LPS-exposed AECII at 6 h and mouse SPC-positive (SPC) cells after 3-day LPS exposure. We observed that downregulated DCLK1 increased p-YAP/YAP, while DCLK1 overexpression slightly reduced p-YAP/YAP, indicating an association between DCLK1 and Hippo-YAP pathway.
We conclude that DCLK1-mediated Hippo signaling components of YAP/TAZ regulated markers of AECII-to-AECI differentiation in an LPS-induced ARDS model.
在急性呼吸窘迫综合征（ARDS）患者中，II 型肺泡上皮细胞（AECII）再生延迟与更高的死亡率有关。然而，双皮质激酶 1（DCLK1）与 Hippo 信号通路在 ARDS 相关的 AECII 分化中的相互作用尚不清楚。因此，本研究的目的是了解 DCLK1/Hippo 通路在介导 ARDS 中 AECII 分化中的作用。
将 AECII MLE-12 细胞暴露于 0、0.1 或 1μg/ml 的脂多糖（LPS）中 6 和 12 小时。在小鼠模型中，将 C57BL/6JNarl 小鼠经气管内（i.t.）注射 0（对照）或 5mg/kg LPS，并在第 3 和 7 天处死取肺组织。
我们发现 LPS 通过减少表面活性蛋白 C（SPC）和 p53，同时增加 T1α（足突蛋白）和 E-钙粘蛋白，在 12 小时诱导 AECII 分化标志物。同时，在 LPS 暴露的 AECII 中，在 12 小时内观察到核 YAP 的动态调节和 TAZ 水平的增加。YAP 抑制一致降低了 SPC、claudin 4（CLDN-4）、半乳糖凝集素 3（LGALS-3）和 p53 的细胞水平，同时在 6 小时增加了跨上皮电阻（TEER）。此外，在 ARDS 的分离人 AECII 中观察到 DCLK1 表达降低，与 LPS 暴露的 AECII 在 6 小时和小鼠 SPC 阳性（SPC）细胞在 3 天 LPS 暴露后的结果一致。我们观察到下调的 DCLK1 增加了 p-YAP/YAP，而 DCLK1 过表达则轻微降低了 p-YAP/YAP，表明 DCLK1 与 Hippo-YAP 通路之间存在关联。
我们的结论是，DCLK1 介导的 Hippo 信号通路 YAP/TAZ 调节了 LPS 诱导的 ARDS 模型中 AECII 向 AECI 分化的标志物。
