PROGENITOR COLONY RT-PCR ANALYSIS IN CML TREATMENT

描述：（申请人的摘要）慢性髓性白血病（CML）是 通常与BCR和ABL之间的相互易位有关 引起BCR/ABL杂合癌蛋白合成的基因。一 将对接受新治疗的CML患者进行详细研究 涉及诱导方案、体内净化、干细胞 动员到外周血中，白细胞去除术，自体外周血 造血干细胞移植和移植后治疗 α-干扰素计划的研究将是一个相关的实验室， 临床研究它将监测骨髓中白血病细胞的存在 在治疗过程的四个关键阶段通过逆转录 然后进行BCR/ABL的聚合酶链反应（RT-PCR） 易位产物最重要的RT-PCR检测要做的是， 研究将是那些使用体外衍生自骨髓单个集落的研究， 用于分析BCR/ABL产物。在CML研究中 仅将未培养的材料用于该RT-PCR的患者或 进行细胞遗传学分析或荧光原位杂交以检测 BCR/ABL易位，大多数测定的细胞不是BCR/ABL易位。 白血病祖细胞在拟议的研究中，将对来自 从骨髓中的祖细胞培养的单个集落， CML患者的外周血样本，通过定性和 竞争性定量RT-PCR，对来自相应的未培养的 样品白血病祖细胞频率的RT-PCR分析结果 还将细胞与常规细胞遗传学分析的细胞进行比较。 这项研究将允许直接比较的污染水平， 入组后外周血样本和骨髓样本中有白血病细胞 体内净化此外，拟议的分析将提供所需的初步数据 进行更大规模的研究，以测试 自体外周血中BCR/ABL阳性髓系祖细胞 用于移植和长期无病生存的干细胞。

DESCRIPTION: (Applicant's Abstract) Chronic myelogenous leukemia (CML) is usually associated with a reciprocal translocation between the BCR and ABL genes that gives rise to synthesis of the hybrid BCR/ABL oncoprotein. A detailed study will be conducted of CML patients undergoing a new treatment protocol involving an induction regimen, in vivo purging, stem cell mobilization into the peripheral blood, leukapheresis, autologous peripheral blood stem cell transplantation and post-transplant treatment with alpha-interferon. The planned research will be a correlative laboratory and clinical study. It will monitor the presence of leukemic cells in bone marrow at four critical stages in the treatment procedure by reverse transcription followed by the polymerase chain reaction (RT-PCR) for the BCR/ABL translocation product. The most important RT-PCR assays to be done in this study will be those that use individual colonies derived in vitro from myeloid progenitor cells for analysis of the BCR/ABL product. In studies on CML patients in which only uncultured material is used for this RT-PCR or in which cytogenetic analysis or fluorescence in situ hybridization is done to detect the BCR/ABL translocation, the majority of the assayed cells are not the leukemic progenitors. In the proposed study, RT-PCR will be done on RNA from individual colonies cultured from progenitor cells in bone marrow and peripheral blood samples from CML patients and, by both qualitative and competitive quantitative RT-PCR, on RNA from the corresponding uncultured samples. The results of RT-PCR analysis of the frequency of leukemic progenitor cells will also be compared to those from conventional cytogenetic analyses. This study will permit a direct comparison of the level of contamination of peripheral blood samples and bone marrow samples with leukemic cells after in vivo purging. Also, the proposed analysis will provide needed preliminary data for a larger study to test the relationship between the levels of the BCR/ABL-positive myeloid progenitor cells in the autologous peripheral blood stem cells used for engraftment and long-term disease-free survival.