Structural and transcriptomic response to antenatal corticosteroids in an Erk3-null mouse model of respiratory distress.
Structural and transcriptomic response to antenatal corticosteroids in an Erk3-null mouse model of respiratory distress.
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DOI：
10.1016/j.ajog.2016.04.043
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发表时间：
2016-09
影响因子：
9.8
通讯作者：
Aagaard K
中科院分区：
文献类型：
--
作者：
Pew BK;Harris RA;Sbrana E;Guaman MC;Shope C;Chen R;Meloche S;Aagaard K
早产儿中的新生儿呼吸窘迫综合征是新生儿死亡的主要原因。通过产前糖皮质激素治疗和新生儿表面活性物质替代治疗，肺功能不全相关的婴儿死亡率有所改善。然而，尽管临床使用了几十年，糖皮质激素促进胎肺成熟的机制仍不完全清楚。我们之前已经证明，Erk3基因缺失会导致小鼠生长受限、发紫和早期新生儿死亡，原因是肺发育不成熟和呼吸窘迫。最近，我们证明了出生后在地塞米松治疗的基础上添加表面活性物质可以提高Erk3基因缺失的新生小鼠的存活率。为了更好地了解皮质类固醇介导的肺成熟的分子基础，我们使用了高通量转录和高分辨率的小鼠胎肺形态分析。我们试图检查与新生儿呼吸窘迫和产前糖皮质激素治疗相关的胎儿肺结构和功能的变化。在胚胎16.5天和17.5天分别给予地塞米松(0.4 mg/kg)或生理盐水。收集胎肺，用MicroCT和RNA-seq分析基因表达的差异以及与基因和治疗的途径相互作用。转录分析的结果指导了利用免疫染色在小鼠和人胎肺组织中进一步研究候选基因。Erk3−/−小鼠表现出肺不张，与野生型相比，总孔隙率和球囊空间减少，经糖皮质激素治疗后得到改善。在rna-seq检测到的596个差异表达基因中，通径分析显示36个基因(q<；0.05)与地塞米松相互作用，其中一些基因与肺发育有关，包括促肾上腺皮质激素释放激素和表面活性蛋白B。在胚胎14.5天，在野生型和Erk3−/−肺中检测到促肾上腺皮质激素释放激素蛋白，并在胚胎第18.5天显著改变表达。产前地塞米松降低胚胎18.5d的野生型和Erk3−/−肺的促肾上腺皮质激素释放激素(分别是0.56倍和0.67倍；P<；.001)。野生型小鼠对糖皮质激素治疗的反应是肺表面活性物质蛋白B增加(P=0.003)。相反，在Erk3−/−小鼠中，地塞米松治疗导致表面活性蛋白B降低(P=0.012)。在人类验证研究中，我们证实促肾上腺皮质激素释放激素蛋白在妊娠18周时存在于胎儿肺中，并随着生存能力的发展而增加表达(妊娠22周；P<；0.01)。对整个转录组基因表达的分析表明，糖皮质激素分别通过Erk3依赖和依赖机制对促肾上腺皮质激素释放激素和表面活性蛋白B进行调节。我们首次证实了促肾上腺皮质激素释放激素蛋白在中期人胎肺中的表达和时间调节。这一独特的模型允许皮质类固醇对胎儿肺形态状况的影响与功能基因途径调节区分开来。这些发现表明，Erk3可能是产前糖皮质激素促进早产儿肺表面活性物质蛋白产生的重要分子介质，并扩大了我们对临床治疗提高新生儿存活率的关键机制的理解。
Neonatal respiratory distress syndrome in preterm infants is a leading cause of neonatal death. Pulmonary insufficiency-related infant mortality rates have improved with antenatal glucocorticoid treatment and neonatal surfactant replacement. However, the mechanism of glucocorticoid-promoted fetal lung maturation is not understood fully, despite decades of clinical use. We previously have shown that genetic deletion of Erk3 in mice results in growth restriction, cyanosis, and early neonatal lethality because of pulmonary immaturity and respiratory distress. Recently, we demonstrated that the addition of postnatal surfactant administration to antenatal dexamethasone treatment resulted in enhanced survival of neonatal Erk3-null mice. To better understand the molecular underpinnings of corticosteroid-mediated lung maturation, we used high-throughput transcriptomic and high-resolution morphologic analysis of the murine fetal lung. We sought to examine the alterations in fetal lung structure and function that are associated with neonatal respiratory distress and antenatal glucocorticoid treatment. Dexamethasone (0.4 mg/kg) or saline solution was administered to pregnant dams on embryonic days 16.5 and 17.5. Fetal lungs were collected and analyzed by microCT and RNA-seq for differential gene expression and pathway interactions with genotype and treatment. Results from transcriptomic analysis guided further investigation of candidate genes with the use of immunostaining in murine and human fetal lung tissue. Erk3−/− mice exhibited atelectasis with decreased overall porosity and saccular space relative to wild type, which was ameliorated by glucocorticoid treatment. Of 596 differentially expressed genes (q < 0.05) that were detected by RNA-seq, pathway analysis revealed 36 genes (q < 0.05) interacting with dexamethasone, several with roles in lung development, which included corticotropin-releasing hormone and surfactant protein B. Corticotropin-releasing hormone protein was detected in wild-type and Erk3−/− lungs at E14.5, with significantly temporally altered expression through embryonic day 18.5. Antenatal dexamethasone attenuated corticotropin-releasing hormone at embryonic day 18.5 in both wild-type and Erk3−/− lungs (0.56-fold and 0.67-fold; P < .001). Wild type mice responded to glucocorticoid administration with increased pulmonary surfactant protein B (P = .003). In contrast, dexamethasone treatment in Erk3−/− mice resulted in decreased surfactant protein B (P = .012). In human validation studies, we confirmed that corticotropin-releasing hormone protein is present in the fetal lung at 18 weeks of gestation and increases in expression with progression towards viability (22 weeks of gestation; P < .01). Characterization of whole transcriptome gene expression revealed glucocorticoid-mediated regulation of corticotropin-releasing hormone and surfactant protein B via Erk3-independent and -dependent mechanisms, respectively. We demonstrated for the first time the expression and temporal regulation of corticotropin-releasing hormone protein in midtrimester human fetal lung. This unique model allows the effects of corticosteroids on fetal pulmonary morphologic condition to be distinguished from functional gene pathway regulation. These findings implicate Erk3 as a potentially important molecular mediator of antenatal glucocorticoid action in promoting surfactant protein production in the preterm neonatal lung and expanding our understanding of key mechanisms of clinical therapy to improve neonatal survival.
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DOI：
10.1152/ajplung.00127.2002
发表时间：
2002-11-01
影响因子：
4.9
作者：
Gonzales, LW;Guttentag, SH;Ballard, PL
通讯作者：
Ballard, PL
影响因子：
2.5
作者：
Dullin C;Larsson E;Tromba G;Markus AM;Alves F
通讯作者：
Alves F
影响因子：
9.8
作者：
Ecker, Jeffrey L.;Kaimal, Anjali;Sciscione, Anthony C.
通讯作者：
Sciscione, Anthony C.
影响因子：
2.5
作者：
Dullin C;dal Monego S;Larsson E;Mohammadi S;Krenkel M;Garrovo C;Biffi S;Lorenzon A;Markus A;Napp J;Salditt T;Accardo A;Alves F;Tromba G
通讯作者：
Tromba G
影响因子：
4
作者：
LIGGINS, GC
通讯作者：
LIGGINS, GC