Studies on the trans-differentiation of hematopoietic stem cells and the application for the gene therapy

造血干细胞转分化研究及其在基因治疗中的应用

• 批准号: 14570975
• 负责人: ITOH Katsuhiko
• 金额: $ 2.24万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570975/
• 关键词: Hematopoietic stem cell; retroviral vector; hepatocyte; hepatocyte; bone marrow; stem cell; retrovirus; vector; レトロウイルスベクター

Recent report indicated that the hematopoietic stem cells can trans-differentiate into hepatocytes, neurons, muscle cells or epitherial cells and these cells are the candidate of the target cells in the gene therapy.We have proved that the transduced hematopoietic stem cells by the retroviral vectors carrying anti-cancer drug resistant gene (MGMT/P14OK) could be enriched in vivo and that the almost 100% of periferal blood cells were replaced by the transduced cells. On the other hand, after the replacement, only a small proportion of the hepatocytes (a few cells per 100,000-1,000,000 cells) was confirmed to be derived from transduced hematopoietic cells. Thus, we have concluded that the proportion of the hematopoietic cells is very low, which would trans-differentiate into hepatocyte in vivo.We have proved that the FMEV-type of the retroviral vectors were superior to those based on MoMLV in the expression of transgene in the liver. Furthermore, we have developed novel vectors (SF-Hep3 and SF-Hep5) by engineering the U3 region of the LTR, which provided efficient transgene expression in hepatocytes in vivo.

最近的研究表明，造血干细胞可以向肝细胞、神经元、肌肉细胞或上皮样细胞反式分化，这些细胞是基因治疗的候选靶细胞。我们已经证明，携带抗癌耐药基因的逆转录病毒载体(MGMT/P14OK)转导的造血干细胞可以在体内得到丰富，几乎100%的外周血细胞被转导的细胞所替代。另一方面，置换后只有一小部分肝细胞(每100,000-1,000,000个细胞中有几个细胞)被证实来自转导的造血细胞。由此可见，FMEV型逆转录病毒载体的造血细胞比例很低，在体内可向肝细胞反式分化，证明了FMEV型逆转录病毒载体在肝脏中的表达优于MoMLV载体。此外，我们还开发了新型载体(SF-Hep3和SF-Hep5)，通过对LTR的U3区域进行工程，提供了在体内肝细胞中高效的转基因表达。

项目成果

Ohnishi N et al.: "High expression of transgenes mediated by hybrid retroviral vectors in hepatocytes : comparison of promoters from murine retroviruses in vitro and in vivo"Gene Ther.. 9. 303-306 (2002)

Ohnishi N 等人：“肝细胞中杂合逆转录病毒载体介导的转基因高表达：体外和体内鼠逆转录病毒启动子的比较”Gene Ther.. 9. 303-306 (2002)

Higashitsuji H et al.: "A novel protein overexpressed in hepatoma accelerates export of NF-kB from the nucleus and inhibits p53-dependent apoptosis"Cancer Cell. 2. 335-346 (2002)

Higashitsuji H 等人：“肝癌中过度表达的一种新型蛋白质可加速 NF-kB 从细胞核的输出，并抑制 p53 依赖性细胞凋亡”癌细胞。

Sakurai T et al.: "A cleaved form of MAGE-A4 binds to Miz-1 and induces apoptosis in human cells"J.Biol.Chem.. (in press).

Sakurai T 等人：“MAGE-A4 的裂解形式与 Miz-1 结合并诱导人类细胞凋亡”J.Biol.Chem..（出版中）。

Sakurai T et al.: "A Cleaved Form of MAGE-A4 Binds to Miz-1 and Induces Apoptosis in Human Cells"J.Biol.chem.. 279. 15505-15514 (2004)

Sakurai T 等人：“A Cleaved Form of MAGE-A4 Binds to Miz-1 and Induces Apoptosis in Human Cells”J.Biol.chem. 279. 15505-15514 (2004)

Friel J, et al.: "Hierarchy of stroma-derived factors in supporting growth of stroma-dependent hemopoictic cells Membrane-bound SCF is sufficient to confer stroma competence to epitherial cells."Growth Factors. 20. 35-51 (2002)

Friel J 等人：“基质衍生因子的层次结构支持基质依赖性造血细胞的生长，膜结合的 SCF 足以赋予上皮细胞基质能力。”生长因子。