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.

硫氧还蛋白 (TRX) 是一种氧化还原调节剂，已知可以减少因使用抗肿瘤药物或放疗引起的氧应激。基于这一知识，我们挑战建立一种新方法来保护骨髓细胞免受化疗引起的细胞损伤。 1998年，在许多造血细胞系中检测了TRX基因的表达，每个细胞的内源性TRX转录物的表达都非常弱。然后将 TRX 基因诱导至 NIH3T3 成纤维细胞，并检查 TRX 转染子的耐药性是否获得，结果显示对 HIID22D2OD22D2、顺铂和 4-羟基环磷酰胺有轻微耐药性，然后将 TRX 基因诱导至造血细胞（包括骨髓细胞）。获得TRX诱导的造血细胞如此困难可能是因为诱导率低1999年，为了克服这一技术难题，我们通过共诱导角质形成细胞生长因子（KGF）受体基因K-sam，即K-sam基因转移至造血细胞系后，成功建立了独特的系统。小鼠骨髓细胞，用K-sam配体KGF刺激细胞，通过该方法，k-sam诱导的细胞被优先扩增。还发现KGF刺激不会发生分化诱导，不会改变细胞特性，因此该方法被认为是一种新型高效的造血干细胞扩增模型，值得临床应用。