GENE THERAPY MOUSE MODEL FOR CML

描述：慢性粒细胞白血病（CML）是一种疾病， 已知初始损伤发生在造血干细胞水平 （HSC）。 超过95%的CML患者的白血病细胞含有 费城（Ph），这是一个互惠的结果， 染色体9和22之间的易位。 在分子水平上， 易位产生bcr/abl杂合基因，其产物增加， 酪氨酸激酶活性。 这些变化与观察到的非常相似 在小鼠中发现的类似改变和激活的v-abl癌基因中。 bcr/abl或v-abl癌基因导入造血系统 干/祖细胞和它们植入受体小鼠后， 产生一种类似人类慢性粒细胞白血病的疾病。 在过去几年中， 包括首席研究员在内的几位研究人员积极地 使用小鼠模型来了解疾病的发病机制。 把它拿 此外，他们正在建立一种慢性粒细胞白血病基因治疗小鼠模型。 通过使用 胎儿肝细胞作为HSC的来源，他们已经表明， 小鼠白血病可以在逆转录病毒转移后发展， bcr/abl或v-abl癌基因进入这些胎儿HSC。 此外，这只老鼠 模型有几个方面与人类相似：（a）未转导 正常的HSC也存在于白血病小鼠中，和（B）正常HSC的表达， Ph +ve人原始干细胞中激活的abl癌基因水平较低 多能干/祖细胞。 在本申请中，研究人员 我希望进一步分析这个模型，并检验逆转录病毒基因转移是否 将抗bcr/abl序列导入白血病造血干细胞会导致细胞生长迟缓， 或消除继发性受者的白血病发展， 单个或少量白血病供体HSC的再移植。 具体目标 是（1）确定bcr/abl转导和未转导的比例 在白血病小鼠中长期重建HSC;（2）检查是否 未转导的gp 105 +ive HSC具有竞争性生长优势， bcr/abl转导的gp 105 +ive HSC;（3）验证 反义核酸载体对bcr/abl介导的细胞凋亡的抑制作用 转化或白血病发展;（4）测试 通过使用体内反义核酸将基因转移到白血病HSC中的反义疗法 小鼠白血病模型。

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.