CHROMOSOMAL ABNORMALITIES IN MYELOMA AS DETECTED BY FISH

鱼检测骨髓瘤染色体异常

• 批准号: 6026912
• 负责人: Rafael Fonseca
• 金额: $ 19.66万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6026912
• 关键词: acute phase protein; aneuploidy; chromosome deletion; chromosome translocation; cyclins; cytokine receptors; diagnosis design /evaluation; fibroblast growth factor; fluorescent in situ hybridization; growth factor receptors; human tissue; interleukin 6; leukemia; major histocompatibility complex; multiple myeloma; neoplasm /cancer diagnosis; neoplasm /cancer genetics; polymerase chain reaction

DESCRIPTION (adapted from the investigator's abstract): Background; Multiple myeloma (MM) is an incurable plasma cell malignancy. Most investigators believe that progression and outcome in MM is secondary to specific genetic alterations of the malignant cells. Translocations at 14q32 are thought to be the genetic hallmark of MM. By conventional cytogenetic analysis (CC), others and Dr. Fonseca have determined that the presence of chromosomal abnormalities is associated with an adverse outcome. Structural abnormalities of the long arm of chromosomes 11 (mostly translocations), and 13 and the short arm of 17 have special prognostic significance. However, CC is fraught with many difficulties including inability to detect abnormalities in non-proliferating cells. Fluorescent in situ hybridization (FISH) can detect chromosomal abnormalities in interphase cells. The prognostic significance of FISH is not yet known. Preliminary data suggest that chromosomal abnormalities as detected by FISH are of significance as well. Understanding the genetic mechanisms of MM cell proliferation, disease progression and outcome is important to eventually overcoming the disease. Furthermore these abnormalities may identify biologically different subgroups of MM. Hypothesis: 1) Specific chromosomal structural and numerical abnormalities will have prognostic significance in patients with MM (overall survival and event free survival). 2) Translocations will result in gene overexpression as detected by RNA analysis (reverse transcriptase polymerase chain reaction, TR-PCR) and immunohistochemistry. 3) These chromosomal abnormalities will have correlations with known biological and prognostic factors. 4) Specific chromosomal abnormalities at two years after initiation of treatment may result in development of therapy-related myelodysplasia or acute leukemia (tMDS/AML). Objectives: 1) Determine the frequency and prognostic significance (overall survival and event free survival) of translocations between 14q32 and other donors chromosomes (11q13, 4q16.3, 16q23), deletions (13q and 17p13) and numerical chromosomal abnormalities (chromosomes 6, 7, 9, 11, 15,17). 2) Correlate the presence of specific chromosomal translocations with resulting gene overexpression by RT-PCR (cyclin D1, FGFR3, MMSET, and c-maf) and immunohistochemistry (cyclin D1 only). 3) Correlate chromosomal abnormalities with tumor biological markers (plasma cell labeling index, B-2 microglobulin, C-reactive protein, soluble IL-6 receptor, DNA content S-phase, DNA aneuploidy, and plasmablastic morphologic). 4) Assess for specific chromosomal abnormalities associated to myelodysphasia and therapy related leukemia and relate these findings to clinical development of tMDS/AML. Material and Methods; To detect chromosomal abnormalities he will perform dual-color FISH using locus-specific and chromosome-specific probes on archival samples of 250 patients collected for E9487 (associated ancillary laboratory trial to clinical trial E9486). To precisely identify monotypic plasma cells he will couple FISH with fluorescent staining for the cytoplasmic immunoglobulin. Additionally, he will do RT-PCR for RNA analysis and immunohistochemistry for protein overexpression (cyclin D1). He will perform biological correlations with other markers, and perform an overall survival and event free survival analysis according to the presence or absence of specific abnormalities. Variables will also be studied using a multiple-variable model to test them as independent predictors.

描述（改编自研究者摘要）：背景;多种 骨髓瘤（MM）是一种不可治愈的浆细胞恶性肿瘤。大多数调查人员认为 MM的进展和结局继发于特定的遗传改变， 的恶性细胞。14 q32的易位被认为是遗传性的。 通过常规细胞遗传学分析（CC），其他人和Dr。 丰塞卡已经确定染色体异常的存在是 与不良后果相关。长臂的结构异常 11号染色体（主要是易位），13号染色体和17号染色体的短臂有 特殊的预后意义。然而，CC充满了许多困难 包括不能检测非增殖细胞中的异常。 荧光原位杂交（FISH）可以检测染色体异常 在间期细胞中。FISH的预后意义尚不清楚。 初步数据表明，通过FISH检测到的染色体异常是 也很重要。了解MM细胞的遗传机制 增殖，疾病进展和结果是重要的， 战胜疾病。此外，这些异常可以识别 MM的生物学不同亚组。 假设：1）特定的染色体结构和数量异常将 在MM患者中具有预后意义（总生存期和事件 自由生存）。2)易位将导致基因过表达， 通过RNA分析（逆转录聚合酶链反应， RT-PCR）和免疫组织化学。3)这些染色体异常 与已知生物学和预后因素的相关性。4)具体 治疗开始后2年的染色体异常可能导致 治疗相关的骨髓增生异常或急性白血病（tMDS/AML）的发展。 目的：1）确定频率和预后意义（总体 生存和无事件生存）之间的易位14 q32和其他 供体染色体（11 q13，4q16.3，16 q23），缺失（13 q和17 p13）和 染色体数目异常（6、7、9、11、15、17号染色体）。（二） 将特定染色体易位的存在与结果 通过RT-PCR检测基因过表达（细胞周期蛋白D1、FGFR 3、MMSET和c-maf）， 免疫组化（仅细胞周期蛋白D1）。3)相关染色体异常 用肿瘤生物学标志物（浆细胞标记指数，B-2微球蛋白， C反应蛋白、可溶性IL-6受体、DNA含量、S期、DNA异倍体、 和浆细胞形态学）。4)评估特定染色体 与骨髓困难和治疗相关白血病相关的异常， 将这些发现与tMDS/AML的临床发展联系起来。 材料和方法：为了检测染色体异常，他将进行 利用基因座特异性和染色体特异性探针对档案进行双色FISH 为E9487（相关辅助实验室）采集的250例患者样本 临床试验E9486）。为了精确识别单型浆细胞， 将FISH与细胞质免疫球蛋白的荧光染色偶联。 此外，他将做RT-PCR的RNA分析和免疫组织化学， 蛋白质过表达（细胞周期蛋白D1）。他将进行生物关联 与其他标志物，并进行总生存期和无事件生存期 根据特定异常的存在或不存在进行分析。 还将使用多变量模型对变量进行研究， 独立预测者