Mouse lung organoid responses to reduced, increased, and cyclic stretch.
Mouse lung organoid responses to reduced, increased, and cyclic stretch.
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小鼠肺类器官对减少、增加和循环拉伸的反应。
DOI：
10.1152/ajplung.00310.2020
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Varisco,BrianM
中科院分区：
文献类型：
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作者：
Joshi,Rashika;Batie,MatthewR;Fan,Qiang;Varisco,BrianM
大多数肺发育发生在周期性牵张的背景下。机械微环境的改变是许多肺部疾病的共同特征，其中先天性膈疝（CDH）和胎儿气管闭塞（FETO，CDH的一种治疗方法）是肺结构、细胞分化和功能变化的极端例子。为了解决细胞培养和体内机械力转导模型的局限性，我们使用出生后5天（PND 5）小鼠肺CD 326阳性细胞和成纤维细胞进行增加，减少和循环应变，开发了两种小鼠肺类器官（mLO）机械力转导模型。在第一个模型中，将mLO暴露于毛喉素（FSK）和/或破坏（DIS），并在20小时时进行评估。FSK、对照和DIS mLO的mLO横截面积分别变化了+59%、+24%和-68%。FSK处理的类器官具有两倍于其他类器官的增殖细胞。在第二个模型中，20小时的10.25%双轴循环应变增加肺间充质细胞谱系的mRNA相比，静态拉伸和无拉伸。循环拉伸增加了TGF-β和整合素介导的信号传导，上游分析表明组蛋白脱乙酰酶、microRNA和长非编码RNA的作用。与无牵张和静态牵张mLO相比，周期性牵张mLO增加了α SMA阳性和αSMA-PDGFRα双阳性细胞。在该PND 5 mLO机械转导模型中，细胞增殖通过静态拉伸增加，并且循环拉伸诱导在出生后肺发育中重要的间充质基因表达变化。
Most lung development occurs in the context of cyclic stretch. Alteration of the mechanical microenvironment is a common feature of many pulmonary diseases, with congenital diaphragmatic hernia (CDH) and fetal tracheal occlusion (FETO, a therapy for CDH) being extreme examples with changes in lung structure, cell differentiation, and function. To address limitations in cell culture and in vivo mechanotransductive models, we developed two mouse lung organoid (mLO) mechanotransductive models usingpostnatal day 5(PND5) mouse lung CD326-positive cells and fibroblasts subjected to increased, decreased, and cyclic strain. In the first model, mLOs were exposed to forskolin (FSK) and/or disrupted (DIS) and evaluated at 20 h. mLO cross-sectional area changed by +59%, +24%, and −68% in FSK, control, and DIS mLOs, respectively. FSK-treated organoids had twice as many proliferating cells as other organoids. In the second model, 20 h of 10.25% biaxial cyclic strain increased the mRNAs of lung mesenchymal cell lineages compared with static stretch and no stretch. Cyclic stretch increased TGF-β and integrin-mediated signaling, with upstream analysis indicating roles for histone deacetylases, microRNAs, and long noncoding RNAs. Cyclic stretch mLOs increased αSMA-positive and αSMA-PDGFRα-double-positive cells compared with no stretch and static stretch mLOs. In this PND5 mLO mechanotransductive model, cell proliferation is increased by static stretch, and cyclic stretch induces mesenchymal gene expression changes important in postnatal lung development.