GENE THERAPY MOUSE MODEL FOR CML

CML 基因治疗小鼠模型

• 批准号: 2712789
• 负责人: PETER M WONG
• 金额: $ 29.87万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2712789
• 关键词: antisense nucleic acid; athymic mouse; cell transformation; chromosome translocation; chronic myelogenous leukemia; disease /disorder model; embryo /fetus cell /tissue; fusion gene; gene expression; gene therapy; genetic transduction; hematopoietic stem cells; laboratory mouse; model design /development; mouse leukemia; oncogenes; transfection; transfection /expression vector

DESCRIPTION: Chronic myelogenous leukemia (CML) is a disorder in which the initial lesion is known to occur at the level of hemopoietic stem cells (HSCs). The leukemic cells of over 95 percent of CML patients harbor the Philadelphia (Ph), which occurred as the result of a reciprocal translocation between chromosomes 9 and 22. At the molecular level, this translocation generates the bcr/abl hybrid gene, whose product has elevated tyrosine kinase activity. These changes are very similar to those observed in a similarly altered and activated v-abl oncogene discovered in the mouse. The introduction of either the bcr/abl or v-abl oncogene into hemopoietic stem/progenitor cells and after their engraftment into recipient mice could produce a disease resembling CML in humans. Over the past few years, several investigators, including the principal investigator, have actively employed a mouse model to understand disease pathogenesis. To take it further, they are establishing a gene therapy mouse model for CML. By using fetal liver cells as a source of HSCs, they have shown that transplantable leukemia in mice can be developed after retroviral transfer of either bcr/abl or v-abl oncogene into these fetal HSCs. Furthermore, this mouse model has several aspects that resemble those in humans: (a) untransduced normal HSCs are also present in leukemic mice, and (b) expression of the activated abl oncogene in primitive stem cells is low in Ph +ve human multipotent stem/progenitor cells. In this application, the investigators wish to dissect the model further and to examine if retroviral gene transfer of the anti-bcr/abl sequences into leukemic HSCs would result in retardation or elimination of leukemia development in secondary recipients after retransplantation of single or few leukemic donor HSCs. The specific aims are (1) to determine the proportion of bcr/abl transduced and untransduced long-term repopulating HSCs in leukemic mice; (2) to examine whether untransduced gp105 +ive HSCs have a competitive growth advantage over bcr/abl transduced gp105 +ive HSCs; (3) to verify the effectiveness of the antisense vector in vitro on the suppression of bcr/abl-mediated transformation or leukemia development; (4) to test the efficacy of the antisense therapy by gene transfer into leukemic HSCs using the in vivo mouse model of leukemogenesis.

描述：慢性粒细胞白血病(CML)是一种疾病，其中 已知最初的损害发生在造血干细胞水平。 (HSCs)。95%以上的慢性粒细胞白血病患者的白血病细胞中含有 费城(Ph)，这是一个互惠的结果 9号和22号染色体之间的易位。在分子水平上，这 易位产生BCR/ABL杂交基因，其产物升高 酪氨酸激酶活性。这些变化与观察到的非常相似 在小鼠中发现了类似的改变和激活的v-abl癌基因。 Bcr/abl或v-abl癌基因导入造血细胞 干细胞/祖细胞，在它们植入受体小鼠后 在人类身上产生一种类似慢性粒细胞白血病的疾病。在过去几年里， 包括首席调查员在内的几名调查人员积极 采用小鼠模型来了解疾病的发病机制。拿着它 此外，他们正在为慢性粒细胞白血病建立基因治疗小鼠模型。通过使用 胎肝细胞作为HSCs的来源，他们已经证明了可以移植 小鼠的白血病可以在逆转录病毒转移后发生 Bcr/abl或v-abl癌基因导入这些胎儿HSCs。此外，这款鼠标 模型有几个方面与人类相似：(A)未转换 白血病小鼠中也存在正常的HSCs，并且(B) Ph+Ve人原始干细胞中活化的ABL癌基因水平较低 多能干细胞/祖细胞。在本申请中，调查人员 希望进一步剖析模型并检查逆转录病毒基因转移 将抗bcr/abl序列导入白血病HSCs会导致迟缓 或消除二次移植后白血病的发展 单个或少数白血病供者造血干细胞的再次移植。具体目标 (1)BCR/ABL换算与非换算比例的确定 长期移植白血病小鼠的造血干细胞；(2)检测 未转导的gp105+ive造血干细胞的生长优势优于 Bcr/abl转导的gp105+ive造血干细胞；(3)验证 反义载体体外抑制bcr/abl介导的细胞增殖 转化或白血病的发展；(4)测试疗效。 体内基因转导白血病HSCs的反义治疗 白血病发生的小鼠模型。