Studies on mechanisms apoptosis and carcinogenesis of hepatocytes in hereditary liver disease and approaches for gene therapy for liver diseases

遗传性肝病肝细胞凋亡、癌变机制研究及肝病基因治疗途径

• 批准号: 11470508
• 负责人: ENDO Fumio
• 金额: $ 8.19万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470508/
• 关键词: liver failure; amino acid metabolism; enzyme defect; cancer; mouse model; apoptosis; チロシン; 尿細管

Hereditary tyrosinemia type I (HTI, fumarylacetoacetate hydrolase deficiency) is characterized by severe liver disease and high incidence for liver carcinomas. In this study, we investigated animal model for HT1 to elucidate mechanisms for carcinogenesis in this disease. Previously, we introduced defective HPD gene into albino lethal mice and successively rescued the phenotype. In the present study, we administered homogentisic acid which lead to liver apoptosis in these mice.Following results are obtained.(i) Apoptosis induced by fumarylacetoacetate in hepatocytes and renal tubular epithelial cells were prevented by the administration of caspase inhibitors.(ii) When cell cycles were investigated, the cells injured by fumarylacetoacetate were arrested at G2M.(iii) Recombinant adenovirus expressing human fumarylacetoacetate hydrolase prevents the liver damage and rescue the mice. In addition, recombinant adeno associated virus expressing human fumarylacetoacetate hydrolase rescues the mice.(iv) Administration of homogentisic acid in the model mice was attempted for development of carcinoma. On histological examinatios, abnormal cells were appeared in the livers however, apparent carcinoma was not confirmed in the present study.

遗传性酪氨酸血症I型（HTI，富马酰乙酰乙酸水解酶缺乏症）的特征是严重的肝脏疾病和肝癌的高发病率。在这项研究中，我们研究了HT1的动物模型，以阐明这种疾病的致癌机制。在此之前，我们将缺陷型HPD基因导入白化病致死小鼠并成功挽救了其表型。在本研究中，我们使用尿黑酸诱导这些小鼠的肝细胞凋亡。(i)半胱天冬酶抑制剂的管理，防止肝细胞和肾小管上皮细胞的富马酰乙酰乙酸诱导的细胞凋亡。(ii)当细胞周期进行调查时，由富马酰乙酰乙酸损伤的细胞被阻滞在G2M。(iii)表达人延胡索酰乙酰乙酸水解酶的重组腺病毒对小鼠肝损伤的保护作用。此外，表达人延胡索酰乙酰乙酸水解酶的重组腺相关病毒拯救了小鼠。(iv)在模型小鼠中施用尿黑酸试图用于癌的发展。在组织学检查中，肝脏中出现异常细胞，但在本研究中未证实明显的癌。

项目成果

Sperandeo M.P., Bassi M.T., Roboni M., Parenti G., Buoninconti A., Manzoni M., Incerti B., Larocca M.R, Racco M.D., Strisciuglio P., Dianzani I., Parini R, Candito M., Endo F., Ballabio A., Andria G., Sebastio G., Borsani G.: "Structure of the SLC7A7 Gene

Sperandeo M.P.、Bassi M.T.、Roboni M.、Parenti G.、Buoninconti A.、Manzoni M.、Incerti B.、Larocca M.R、Racco M.D.、Strisciuglio P.、Dianzani I.、Parini R、Candito M.、Endo F.

Sperandeo M.P.: "Structure of the SLC7A7 Gene and Mutational Analysis of Patients Affected by Lysinuric Protein Intolerance."Am J.Hum.Genet.. 66. 92-99 (2000)

Sperandeo M.P.：“SLC7A7 基因的结构和受赖氨酸尿蛋白不耐受影响的患者的突变分析”Am J.Hum.Genet.. 66. 92-99 (2000)

Sun M-S.Hattori S.Kubo S.Awata H.Matsuda I.Endo F.: "A mouse model of renal tubular injury of tyrosinemia type 1 : Development of de Toni Fanconi syndrome and apoptosis of renal tubular cells in Fah/Hpd double mutant mice."J Am Soc Nephrol.. 11. 291-300 (

Sun M-S.Hattori S.Kubo S.Awata H.Matsuda I.Endo F.：“1 型酪氨酸血症肾小管损伤的小鼠模型：Fah/Hpd 双突变体中 de Toni Fanconi 综合征的发展和肾小管细胞凋亡

Sun M-S.: "A mouse model of renal tubular injury of tyrosinemia type 1 : Development of de Toni Fanconi syndrome and apoptosis of renal tubular cells in Fah/Hpd double mutant mice"J Am Soc Nephrol. 11. 291-300 (2000)

Sun M-S.：“酪氨酸血症1型肾小管损伤的小鼠模型：Fah/Hpd双突变小鼠中德托尼范科尼综合征的发展和肾小管细胞凋亡”J Am Soc Nephrol。

Tomoeda K.: "Mutations in the 4-Hydroxyphenylpyruvic Acid Dioxygenase Gene Are Responsible for Tyrosinemia Type III and Hawkinsinuria."Molecular Genetics and Metabolism. 71. 506-510 (2000)

Tomoeda K.：“4-羟基苯基丙酮酸双加氧酶基因的突变导致 III 型酪氨酸血症和霍金斯尿症。”分子遗传学和代谢。