Molecularbiological analysis of human fetal lung development and its clinical application

人胎肺发育的分子生物学分析及其临床应用

• 批准号: 09671196
• 负责人: IKEDA Kazushige
• 金额: $ 1.98万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671196/
• 关键词: TTF-1; HNF-3; intrauterine MRI; SSFSE; nitrofen; fetal lung; pulmonary hypoplasia; SP-B; サーファクタント蛋白; 肺上皮細胞; 肺サーファクタント; 肺の発達

1. TTF-1 is a prerequisite transcription factor for intrauterine lung development, We analyzed cis-element of human TTF-1 gene. 3 kb of TTF-15' region is required for lung selective expression of TTF-1 gene, and HNF-3β upregulate TTF-1 gene transcription in vitro, We made 3 kb human TTF-1 promoted luciferase transgenic mice. Luciferase activity was detected in both lung and brain, This finding shows that 3 kb of TTF-15' region is not enough for lung specific expression in vivo,2. We presented a case of twin who showed a marked difference in signal intensity of the lung on MRI, which was useful for predicting the fetal pathophysiology, Intrauterine MRI using SSFSE provides the possibility of diagnosing hypoplastic lungs prenatally.3. Nitrofen (2,4-dichlorophenyl-p-nitrophenyl ether) causes fetal lung hypoplasia without cingenital diaphragmatic defects, when fed to pregnant mice to Day 9 of gestation. Intratracheal administration of perfluorocarbon resulted in lung expansion and prolonge … More d survival of nitrofen induced lung hypoplasia. Perfluorocarbon may be useful in stabilizing critically in infants with pulmonary hypoplasia during the early phase of their therapy.4. We assessed maturation of pulmonary epithelial cells of human fetuses at 21 weeks of gestation. Immunohistochemical analysis showed SP-B proprotein was detected both bronchioles and terminal airways. However, mature SP-B peptide positive cells were very few. Transmission electron microscopy showed intracellular glycogen granules were still rich and lamellar bodies were few. These findings means the processing from SP-B preproprotein to mature peptide is immature at 21 weeks of gestation.5. To determine whether SP-B protects mice from oxygen-induced injury, heterozygous SP-B+/ gene-targeted mice and wild-type SP-B+/+ littermates were exposed to hyperoxia or room air. Although specific lung compliance in room air in SP-B+/ mice was slightly reduced as compared with that in SP-B+/+ mice, it was reduced more markedly during hyperoxia. Increased alveolar-capillary leakage and relative deficiency of SP-B may contribute to oxygen-induced pulmonary dysfunction in SP-B+/ mice. These data support the concept that SP-B plays an important protective role in the lung. Less

1. TTF-1是子宫内肺发育的先决转录因子，我们分析了人TTF-1基因的顺式元件。TTF-1基因的肺选择性表达需要3 kb的TTF-15′区，并且HNF-3β在体外上调TTF-1基因的转录，我们构建了3 kb的人TTF-1促荧光素酶转基因小鼠。在肺和脑中均检测到荧光素酶活性，这表明3 kb的TTF-15'区域不足以在体内进行肺特异性表达2。我们报告了一对双胞胎的病例，在MRI上显示肺部信号强度的显著差异，这有助于预测胎儿的病理生理，使用SSFSE的宫内MRI提供了产前诊断肺发育不良的可能性。硝基芬（2,4-二氯苯-对硝基苯基醚）给妊娠小鼠至妊娠第9天，可导致胎儿肺发育不全，无先天性膈缺损。气管内灌注全氟化碳可使肺扩张，延长肺发育不全患者的生存期。在肺发育不全的婴儿治疗的早期阶段，全氟碳化合物可能对稳定病情至关重要。我们评估了人类胎儿在妊娠21周时肺上皮细胞的成熟情况。免疫组化分析显示细支气管和终末气道均检测到SP-B蛋白。而成熟的SP-B肽阳性细胞极少。透射电镜显示细胞内糖原颗粒丰富，片层体较少。这些结果表明，在妊娠21周时，SP-B前蛋白向成熟肽的加工尚不成熟。为了确定SP-B是否能保护小鼠免受氧致损伤，我们将SP-B+/基因靶向的杂合子小鼠和SP-B+/+野生型鼠仔暴露于高氧或室内空气中。虽然与SP-B+/+小鼠相比，SP-B+/+小鼠在室内空气中的特定肺顺应性略有降低，但在高氧状态下降低更为明显。SP-B+/小鼠肺泡毛细血管渗漏增加和SP-B相对缺乏可能导致氧诱导的肺功能障碍。这些数据支持SP-B在肺中起重要保护作用的概念。少

项目成果

Ikeda K, Shaw-White JR Wert SE, Whitsett JA: "Hepatocyte nuclear factor 3 activates transcription of thyroid transcription factor 1 in respiratory epithelial cells"Molecular and Cellular Biology. 16. 3626-3636 (1996)

Ikeda K、Shaw-White JR Wert SE、Whitsett JA：“肝细胞核因子 3 激活呼吸道上皮细胞中甲状腺转录因子 1 的转录”分子和细胞生物学。

Tokieda K, Whitesett JA, Clark JC, Weaver TE, Ikeda K, McConnell KB, Jobe Ah, Ikegami M, Iwamoto HS: "Pulmonary dysfunction in neonatal SP-B-deficient mice"American J. of Physiology. 237. L875-L882 (1997)

Tokieda K、Whitesett JA、Clark JC、Weaver TE、Ikeda K、McConnell KB、Jobe Ah、Ikegami M、Iwamoto HS：“新生儿 SP-B 缺陷小鼠的肺功能障碍”美国生理学杂志。

Ikeda K, Yoshihashi H, Kosyo T, Mori K, Hayashida S, Tokieda K, Tanaka M, Miyakoshi K, Fukuzawa R: "Immuohistochemical analysis of thyroid transcription -1(TTF- 1) expression in fetal pulmonary epithelial cells - comparison with SP-B and SP-C -"Acta. Neon

Ikeda K、Yoshihashi H、Kosyo T、Mori K、Hayashida S、Tokieda K、Tanaka M、Miyakoshi K、Fukuzawa R：“胎儿肺上皮细胞中甲状腺转录-1（TTF-1）表达的免疫组织化学分析 - 与 SP 比较

Tokieda K, Whitsett JA, Clark JC, Weaver TE, Ikeda K, McConnell KB, Jobe AH, Ikegami M, Iwamoto HS: "Pulmonary dysfunction in neonatal SP-B-deficient mice."Am J Physiol. 273(4Pt 1 ). L875-L882 (1997)

Tokieda K、Whitsett JA、Clark JC、Weaver TE、Ikeda K、McConnell KB、Jobe AH、Ikegami M、Iwamoto HS：“新生 SP-B 缺陷小鼠的肺功能障碍。”Am J Physiol。

Tokieda K, Iwamoto HS, Bachurski C, Wert SE, Hull WM, lkeda K, Witsett JA: "SP-B deficient mice are susceptible to hyperoxic injury."American J of Respiratory Cell and Molecular Biology. 21(4). 463-472 (1999)

Tokieda K、Iwamoto HS、Bachurski C、Wert SE、Hull WM、lkeda K、Witsett JA：“SP-B 缺陷小鼠易受高氧损伤。”美国呼吸细胞与分子生物学杂志。