In vivo gene transfer study for pathogenesis of lung injury. Endothelin and development of obliterative bronchiolitis

肺损伤发病机制的体内基因转移研究。

• 批准号: 09671378
• 负责人: TAKEDA Shin-ichi
• 金额: $ 2.43万
• 依托单位: Osaka 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 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671378/
• 关键词: In vivo gene transfer; Obliterative bronchiolitis; Endothelin-1(ET-1); Lung transplantation; 生体内遺伝子導入; Enclothelin(ET); Bronchiolitis obliterans; Gene Transfer; HVJ; Lung transplantation; Endothelin; Bronchiolitis Obliterans; Gene Trasnfer

Background. Obliterative bronchiolitis (OB) is a lesion that results when injury to small conducting airways is repaired by a proliferation of fibrous granulation tissue. OB has emerged as a main cause of morbidity and mortality in the setting of lung and heart-lung transplantation. Endothelin-1 (ET-1) initially discovered as a vasoconstrictive peptide, has a mitogenic activity on vascular smooth cells and airway epithelial cells. Overproduction of endothelin has been reported in patients with OB or chronic rejection after lung transplantation. It is still undetermined whether locally overexpressed ET-1 has a potential impact in the pathogenesis of OB. Methods. We locally overexpressed ET-1 using ultraviolet irradiation-inactivated hemagglitinating virus of Japan (HVJ)-liposome-mediated in vivo gene transfer. Plasmid DNA of prepro-ET-1 and high mobility group 1 (HMG1) protein were co-encapsulated in liposomes, and were introduced into airway epithelial cells by HVJ-mediated membrane fu … More sion. Control animals received instillation of HVJ-liposome with an empty expression cassette. To confirm the efficiency of transfection, HVJ liposome with β-galactosidase gene was introduced. The expression of ET-1 and β-galactosidase was assessed by histochemistry. Results. Bronchial epithelium alveolar cells and alveolar macrophage were stained blue (X-Gal) 1 week after in vivo gene transfer of β-gene, indicating β-gal activity. In animals 1 to 2 weeks after in vivo transfection of prepro-ET-1 gene, hyperplastic connective tissue plaque was seen in the alveolar duet and small conducting airway, indicating histologically distinctive bronchiolitis obliterans. Strong ET-1 like immunoactivies were seen in the airway epithelial, hyperplastic connective tissue and alveolar cells. No histopathologic changes were seen in the control animals. Summary and Future Direction. This result first suggests that ET-1 may have an important role in pathogenesis of OB presumably in the "common final pathway". However, the mechanism of mitogenic signaling by ET-1, its regulation of fibroblast proliferation, and its interaction with other cytokines or growth factors remained to be clarified at present. Further studies are needed to ascertain the specific role of ET-1 in the progression of OB. In summary, we conclude that local overexpression of ET-1 has a key role in the pathogenesis of OB. This evidence suggests the potential clinical implication that a pharmacological antagonist or inhibitor may be able to control the progression of this disease in patients with OB. Less

背景。闭塞性细支气管炎（OB）是由纤维性肉芽组织增生修复小导气管损伤引起的一种病变。OB已成为肺和心肺移植中发病和死亡的主要原因。内皮素-1 （Endothelin-1, ET-1）最初被发现是一种血管收缩肽，对血管平滑细胞和气道上皮细胞具有促有丝分裂活性。在OB患者或肺移植后的慢性排斥反应中，内皮素的过量产生已被报道。局部过表达的ET-1是否对OB的发病机制有潜在影响尚不清楚。我们利用紫外线照射灭活日本血凝病毒（HVJ）脂质体介导的体内基因转移，局部过表达ET-1。将pre - et -1质粒DNA与高迁移率组1 （HMG1）蛋白共包被脂质体，通过hvj介导的膜膜注入气道上皮细胞。对照动物用空表达盒灌注hvj脂质体。为了验证转染的有效性，引入了带有β-半乳糖苷酶基因的HVJ脂质体。组织化学法检测ET-1和β-半乳糖苷酶的表达。结果。β-基因在体内转移1周后，支气管上皮肺泡细胞和肺泡巨噬细胞被染成蓝色（X-Gal），表明β-gal有活性。在动物体内转染pre - et -1基因1 ~ 2周后，在肺泡二重和小导气管中可见增生的结缔组织斑块，提示组织学上独特的闭塞性细支气管炎。在气道上皮细胞、增生性结缔组织和肺泡细胞中可见强烈的ET-1样免疫活性。对照组动物未见组织病理学改变。总结和未来方向。这一结果首次提示ET-1可能在OB的发病机制中发挥重要作用，可能是在“共同最终途径”中。然而，ET-1的有丝分裂信号传导机制、对成纤维细胞增殖的调控以及与其他细胞因子或生长因子的相互作用目前尚不清楚。需要进一步的研究来确定ET-1在OB进展中的具体作用。总之，我们得出结论，ET-1的局部过表达在OB的发病机制中起关键作用。这一证据表明，潜在的临床意义是，药物拮抗剂或抑制剂可能能够控制OB患者的疾病进展

项目成果

S.Takeda et al: "Endothelin and lung disease models" J Carodiovascular Pharmacology. 31. S336-S338 (1998)

S.Takeda 等人：“内皮素和肺部疾病模型”《心血管药理学杂志》。

S.Takeda et al: "In vivo assasant of change ain and tissue volume after precomonatoy" J Applied Physiology. 82. 1340-1348 (1997)

S.Takeda 等人：“预生后变化和组织体积的体内分析”J Applied Physiology。

Shin-ichi_Takeda, Yoshiki Sawa, Masato Minami, Yasufumi Kaneda, Yoshitaka Fujii, Ryota Shirakura, Masashi Yanagisawa, Hikaru Matsuda: "Experimental bronchiolitis obliterans induced by in vivo HVJ liposome-mediated endothelin-1 gene transfer"Ann Thorac Sur

Shin-ichi_Takeda、Yoshiki Sawa、Masato Minami、Yasufumi Kaneda、Yoshitaka Fujii、Ryota Shirakura、Masashi Yanagisawa、Hikaru Matsuda：“体内 HVJ 脂质体介导的内皮素 1 基因转移诱导的实验性闭塞性细支气管炎”Ann Thorac Sur

Shin-ichi_Takeda, Shinichiro Miyoshi, Ken-ichi Omori, Tomoki Utsumi, Shigetoyo Kogaki, Yoshiki Sawa, Masashi Yanagisawa Hikara Matsuda.: "Pulmonary disease models induced by in vivo hemagglutinating virus of Japan liposome-mediated endothelin-1 gene trans

Shin-ichi_Takeda、Shinichiro Miyoshi、Ken-ichi Omori、Tomoki Utsumi、Shigetoyo Kogaki、Yoshiki Sawa、Masashi Yanagisawa Hikara Matsuda.：“日本脂质体介导的内皮素-1基因转染体内血凝病毒诱导的肺部疾病模型

S.Takeda et al: "Experimental branchiolitis induced..." Amals of Thoratic Surgery. 65. 1562-1567 (1997)

S.Takeda 等人：“实验性支气管炎诱发……”胸外科 Amals。