Establishment of Novel Method for Hematopoietic Stem Cell Transplantation Using Thioredoxin-Gene Transfer Technique

硫氧还蛋白基因转移技术建立造血干细胞移植新方法

基本信息

批准号：
10670967
负责人：
HATTORI Yutaka
金额：
$ 0.64万
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：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670967/
关键词：
thioredoxin hematopoietic stem cell transplantation gene therapy drug resistance

项目摘要

Thioredoxin (TRX), a redox regulator, is known to reduce oxygen stress which is caused by the use of antineoplastic drug or irradiation. Based on this knowledge, we challenged to establish a novel method to protect bone marrow cells from cell damage caused by chemotherapy or irradiation therapy by TRX-gene transfer. In 1998, TRX-gene expression was examined in a number of hematopoietic cell lines. Every cell expressed very weakly the endogenous TRX-transcripts. TRX gene was then induced to NIH3T3 fibroblast and was examined acquisition of drug resistance. TRX transfectants were cloned and revealed slight resistance to HィイD22ィエD2OィイD22ィエD2, cisplatinum and 4-hydroxycyclofosfamide. TRX-gene was then induced to hematopoietic cells including bone marrow cells. However, it was so difficult to obtain TRX-induced hematopoietic cells probably because of low induction efficiency. In 1999, to overcome this technical difficulty we successfully established unique system by co-induction of keratinocyte growth factor (KGF) receptor-gene, K-sam. Namely, after K-sam-gene trdansfer to hematopoietic cell lines as well as murine bone marrow cells, the cells were stimulated with K-sam-ligand, KGF. By this method, k-sam-induced cells were preferentially amplified. Incidentally, it was also found that KGF stimulation did not occur induction of differentiation and did not alter characteristics of the cells. Therefore, this method is considered to be a novel efficient model for hematopoietic stem cell expansion, and the clinical application is expected.

已知氧化还原调节剂硫氧还蛋白（TRX）可减少氧气应激，这是由于使用抗塑性药物或辐射引起的。基于这些知识，我们挑战了一种新的方法，以保护骨髓细胞免受化学疗法或通过TRX-GENE转移的辐射治疗引起的细胞损伤。 1998年，在许多造血细胞系中检查了TRX基因表达。每个细胞都非常微弱地表达了内源性TRX transscripts。然后将TRX基因诱导到NIH3T3成纤维细胞中，并检查耐药性的获得。克隆TRX转化体，并显示出对H22E D2OII D22E D2，顺铂和4-羟基环氯氯化物胺的耐药性。然后将TRX基因诱导到包括骨髓细胞在内的造血细胞。但是，很难获得TRX诱导的造血细胞，这可能是由于诱导效率低。在1999年，为了克服这一技术困难，我们通过共同诱导角质形成细胞生长因子（KGF）受体基因（K-SAM）成功建立了独特的系统。也就是说，在k-sam-gene trdansfer到造血细胞系以及鼠骨髓细胞后，用k-SAM-ligand kgf刺激细胞。通过这种方法，优选地将K-SAM诱导的细胞放大。顺便说一句，还发现KGF模拟没有发生分化的诱导，也没有改变细胞的特征。因此，该方法被认为是造血干细胞扩展的新型有效模型，并且预计临床应用。