CORE--HYPERMETAPHASE FISH AND PCR IN ACUTE MYELOGENOUS LEUKEMIA

核心--急性髓系白血病的超中期 Fish 和 PCR

• 批准号: 6237232
• 负责人: MICHAEL J SICILIANO
• 金额: $ 15.12万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-27 至 1998-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6237232
• 关键词: acute myelogenous leukemia; biomedical facility; bone marrow; cell cycle; chromosome aberrations; fluorescent in situ hybridization; human tissue; neoplasm /cancer diagnosis; neoplasm /cancer genetics; polymerase chain reaction; prognosis; tissue /cell culture

A fluorescence in-situ hybridization (FISH) method will be further developed and utilized as a major procedure for the high resolution quantitation of the frequency of cycling AML cells. AML diseases associated with the following chromosomal abnormalities will be studied by this procedure - inv(16), t8;21, t15;17, +8, and -7. The procedure is based on our results in CML where we have coupled long term mitotic arrest to produce thousands of metaphases/slide ("hypermetaphase") with the use of a FISH probe that detects the chromosomal rearrangement associated with the malignancy (hypermetaphase/FISH or HMF). In the CML model we have demonstrated that cancer cells could be readily quantitated in cycling cells from patient bone marrow preparations. The procedures will allow detection of <1% cancer, calls and will permit monitoring of <4% changes in the frequency of such cells. The methodology will be applied here to identify severity of disease in AML patients at diagnosis and to evaluate the effectiveness of therapies on AML patients during treatment. The data from +8 and -7 will be correlated with the interphase FISH data on those same samples generated by Core C. In addition we will conduct reverse transcription followed by PCR (RT/PCR) to determine the level of chimeric transcript in t15;17 and t8;21 AMLs to correlate with the HMF data as well as the interphase FISH and standard G-band cytogenetics (CG) data generated on the same samples by Core Finally we will complete development and conduct genomic PCR on inv(16) samples to evaluate the level of cells containing the chimeric gene reactive to our HMF results as well as RT/PCR, interphase FISH and CG conducted on those same samples. The experiments and correlations will be evaluated with the progress of the patients determined by Projects 1 and 2 (Drs.Estey and Champlin respectively) to determine the benefits and shortcomings of each of the technologies in establishing the cytogenetic response of patients to therapies involved. These procedures will give us new insights into determining the clinically significant level of residual disease, and guide the further management of individual malignancies in AML patients.

荧光原位杂交（FISH）方法将进一步 作为高水平的主要程序而开发和利用 循环 AML 细胞频率的分辨率定量。反洗钱 与以下染色体异常相关的疾病 将通过此过程进行研究 - inv(16), t8;21, t15;17, +8, 和-7。该程序基于我们在 CML 中的结果，其中我们有 加上长期的有丝分裂停滞，产生了数千个 使用 FISH 探针进行中期/载玻片（“超中期”） 检测与相关的染色体重排 恶性肿瘤（超中期/FISH 或 HMF）。在 CML 模型中，我们有 证明癌细胞可以很容易地定量 从患者骨髓制剂中循环细胞。这 程序将允许检测 <1% 的癌症，电话和意愿 允许监测此类细胞频率的 <4% 变化。 该方法将应用于此处以确定问题的严重性 AML 患者在诊断时患有疾病并评估 治疗期间 AML 患者的治疗效果。这 +8 和 -7 的数据将与间期 FISH 相关 由 Core C 生成的相同样本的数据。此外，我们 将进行逆转录，然后进行 PCR (RT/PCR) 确定 t15;17 和 t8;21 中嵌合转录物的水平 AML 与 HMF 数据以及间期 FISH 相关 和在同一台上生成的标准 G 带细胞遗传学 (CG) 数据 Samples by Core 最后我们将完成开发并进行 对 inv(16) 样本进行基因组 PCR 评估细胞水平 还含有对我们的 HMF 结果有反应的嵌合基因 对相同样品进行 RT/PCR、间期 FISH 和 CG。 实验和相关性将通过 项目 1 和 2 确定的患者进展情况（Drs.Estey 和 Champlin 分别）来确定收益和 每种技术在建立 患者对相关治疗的细胞遗传学反应。这些 程序将为我们提供新的见解来确定 残留疾病的临床显着水平，并指导 进一步管理 AML 患者的个体恶性肿瘤。