PROGENITOR COLONY RT-PCR ANALYSIS IN CML TREATMENT

描述：(申请人摘要)慢性粒细胞白血病(CML)是 通常与BCR和ABL之间的相互易位有关 导致合成BCR/ABL杂合癌蛋白的基因。一个 将对接受新治疗的CML患者进行详细研究 方案包括诱导方案、体内净化、干细胞 外周血动员、白细胞分离、自体外周血 血液干细胞移植和移植后的治疗 α-干扰素。计划中的研究将是一个相关的实验室和 临床研究。它将监测骨髓中白血病细胞的存在 在治疗过程中的四个关键阶段通过逆转录 然后用聚合酶链式反应(RT-PCR)检测bcr/abl 易位产物。在这一过程中最重要的RT-PCR分析 研究将是那些使用从髓系细胞体外获得的单个克隆的研究 用于分析bcr/abl产物的祖细胞。在慢性粒细胞白血病的研究中 仅使用未培养的材料进行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，大多数被检测的细胞不是 白血病的祖细胞。在拟议的研究中，RT-PCR将对来自 从骨髓祖细胞培养的单个集落和 慢性粒细胞白血病患者的外周血样本，通过定性和 竞争性定量RT-PCR，对来自相应未培养细胞的RNA 样本。白血病前体细胞基因频率的RT-PCR分析结果 细胞也将与传统细胞遗传学分析中的细胞进行比较。 这项研究将使我们能够直接比较 入院后外周血和骨髓标本中有白血病细胞 活体净化。此外，拟议的分析将提供所需的初步数据 进行一项更大规模的研究，以测试体内 自体外周血中bcr/abl阳性的髓系祖细胞 干细胞用于植入和长期无病生存。

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.