Studies on mechanisms for apoptosis and carcinogenessis, and stam cell transplantation in hereditary liver diseases.

研究细胞凋亡和致癌机制，以及遗传性肝病的干细胞移植。

• 批准号: 13470508
• 负责人: ENDO Fumio
• 金额: $ 9.28万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470508/
• 关键词: liver failure; amino acid metabolism; enzyme defect; cancer; mouse model; apoptosis; 遺伝性疾患; 肝細胞; 遺伝子発現; 遺伝子チップ

Hreditary tyrosinemia typelis a genetic disease caused by deficiency of fumarylacetoacetate hydrolase (FAH). The patients show severe visceral injuries. Mice with FAH deficiency are neonatal leathal and this hampered the efforts to elucidate the mechanisms of disease process. The use of Fah-/-Hpd-/- double mutants provided new opportunity for studies on tyrosinemias. The studies using the models revealed essential features of visceral injury in this disease.(1) We analyzed altered expression of genes after the administration of homogentisate in the double mutants by using gene chips. In thisexperiment gene expression patterns in the livers of double mutants and III mice (HPD-/-) were compared. The RNAs interested were analyzed by real-time quantitative RT-PCR Reduction of expression of metabolic enzymes related to carbohydrate, fatty acids are prominent. These results imply that in the patients with HT1, abnormality of metabolisms and blood coagulation factors are specific events relat … More ed to accumulation of FAA in hepatocytes but not one aspect of general liver damage.(2) we investigated possibility for the presence of endodermal progenitor cells in the damaged salivary gland. For this experiments we used FAH deficient mice as a donor for cell transplantations. After intrasplenic injection of cells prepared from damaged salivary glands, the donor cells were found the liver. Finally we isolated epithelium like cells which proliferate on type I collagen-coated dishes.(3) The progenitor cell expresses both a6b1 integrin and cytoplasmic laminin. Neural progenitor marker nestin, hematopoietic stem cell marker CD34, and hepatic oval cell markers such as albumin, afetoprotein (AFP) and cytokeratin 19 are negative in this cell. When the cells were transplanted into the liver via portal vein, cells wereintegrated into hepatic trabecula and produced albumin.(4) When SGP-1 cells formed clusters on type I collagen-coated dishes, they differentiated into endodermal lineage and two major types of clusters appeared. One consisted of hepatocyte-like cell type (hepatic cluster) and the other consisted of islet-like cell type of the pancreas (pancreatic cluster). The hepatic clusters contained cells positive for AFP and/or albumin, and the cells in the pancreatic clusters are positive for glucagon and/or insulin. These cells aretissue stem cells and able to differentiate into endodermal lineageincluding hepatocyte-like cells and islet-like cells. Less

遗传性酪氨酸血症是一种由富马酰乙酰乙酸水解酶（FAH）缺乏引起的遗传性疾病。病人的内脏有严重损伤。FAH缺乏的小鼠是新生的，这阻碍了阐明疾病过程机制的努力。Fah-/-Hpd-/-双突变体的应用为酪氨酸血症的研究提供了新的契机。使用模型的研究揭示了这种疾病中内脏损伤的基本特征。(1)我们用基因芯片分析了尿黑酸处理后双突变体基因表达的改变。在本实验中，我们比较了双突变型和III型（HPD-/-）小鼠肝脏的基因表达模式。通过实时定量RT-PCR分析感兴趣的RNA。与碳水化合物、脂肪酸相关的代谢酶的表达减少是突出的。这些结果提示，在HT 1患者中，代谢和凝血因子异常是与高血压相关的特异性事件。 ...更多信息 艾德与FAA在肝细胞中的蓄积有关，但不是一般肝损伤的一个方面。(2)我们研究了受损唾液腺中存在内胚层祖细胞的可能性。对于该实验，我们使用FAH缺陷型小鼠作为细胞移植的供体。将从受损的唾液腺制备的细胞脾内注射后，在肝脏中找到了供体细胞。最后，我们分离出上皮样细胞，其在I型胶原包被的皿上增殖。(3)祖细胞表达α 6 β 1整合素和细胞质层粘连蛋白。神经祖细胞标记物巢蛋白、造血干细胞标记物CD 34和肝卵圆细胞标记物如白蛋白、甲胎蛋白（AFP）和细胞角蛋白19在该细胞中为阴性。经门静脉移植入肝后，细胞与肝小梁结合并产生白蛋白。(4)当SGP-1细胞在I型胶原包被的培养皿上形成簇时，它们分化成内胚层谱系，并且出现两种主要类型的簇。一种由肝细胞样细胞类型（肝簇）组成，另一种由胰腺的胰岛样细胞类型（胰腺簇）组成。肝簇含有AFP和/或白蛋白阳性细胞，胰腺簇中的细胞为胰高血糖素和/或胰岛素阳性。这些细胞是组织干细胞，能够分化为内胚层细胞，包括肝细胞样细胞和胰岛样细胞。少

项目成果

Endo F.et al.: "Animal models for tyrosinemias reveal pathophysiologies of tyrosinemias"J. Nutrition. (in press).

Endo F.等人：“酪氨酸血症动物模型揭示了酪氨酸血症的病理生理学”J.

Ishibashi F., Mizukami T,. Kanegasaki S., Motoda L., Kakinuma R., Endo F., Nunoi H.: "Improved superoxide-generating ability by interferon γ due to splicing pattern change of transcripts neutrophils from patients with a splice site mutation in CYBB gene"B

Ishibashi F.、Mizukami T.、Kanegasaki S.、Motoda L.、Kakinuma R.、Endo F.、Nunoi H.：“由于剪接患者的转录物中性粒细胞的剪接模式改变，干扰素 γ 提高了超氧化物生成能力CYBB基因位点突变"B

Matsubasa T., Uchino T., Karashima S., Tanimura M., Endo F.: "Oxidative stress in very low birth weight infants as measured by urinary 8-OhdG"Free Radical Res.. 36. 189-193 (2002)

Matsubasa T.、Uchino T.、Karashima S.、Tanimura M.、Endo F.：“通过尿液 8-OhdG 测量极低出生体重婴儿的氧化应激”自由基研究 36. 189-193 (2002)

Nunoi H.: "Improved superoxide-generating ability by interferon g due to splicing pattern change of transcripts neutrophils from patients with a splice site mutation in CYBB gene"Blood. 98. 436-441 (2001)

Nunoi H.：“由于 CYBB 基因剪接位点突变患者的中性粒细胞转录物剪接模式改变，干扰素 g 提高了超氧化物生成能力”血液。

Endo F., Sun M-S.: "Tyrosinaemia type I and apoptosis of hepatocytes and renal tubular cells"J. Inheril. Metab. Dis.. 25. 227-234 (2002)

Endo F.，Sun M-S.：“I 型酪氨酸血症与肝细胞和肾小管细胞的凋亡”J。