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（根据调查员的摘要改编）：背景；多种的 骨髓瘤（MM）是一种无法治愈的浆细胞恶性肿瘤。大多数调查人员都相信 MM中的进展和结果是特定遗传改变的继发 恶性细胞。 14q32的易位被认为是遗传 MM的标志。通过常规的细胞遗传学分析（CC），其他人和博士 fonseca已经确定染色体异常的存在是 与不利结果相关。长臂的结构异常 染色体11（主要是易位），13个短臂有17个 特殊的预后意义。但是，CC充满了许多困难 包括无法检测非增殖细胞异常。 荧光原位杂交（FISH）可以检测染色体异常 在相间细胞中。鱼类的预后意义尚不清楚。 初步数据表明，鱼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，疾病进展和结局对于最终 克服疾病。此外，这些异常可能会发现 MM的生物学不同亚组。 假设：1）特定的染色体结构和数值异常将 在MM患者中具有预后意义（总生存和事件 免费生存）。 2）易位会导致基因过表达 通过RNA分析检测（逆转录酶聚合酶链反应， TR-PCR）和免疫组织化学。 3）这些染色体异常将具有 与已知的生物学和预后因素相关。 4）具体 启动治疗后两年的染色体异常可能会导致 与治疗相关的骨髓增生或急性白血病（TMDS/AML）开发。 目标：1）确定频率和预后意义（总体 14Q32与其他人之间的易位生存和无事件生存） 供体染色体（11q13，116.3，16q23），删除（13q和17p13）和 数值染色体异常（染色体6、7、9、11、15,17）。 2） 将特定染色体易位的存在与结果相关联 RT-PCR（Cyclin D1，FGFR3，MMSET和C-MAF）和C-MAF的基因过表达和 免疫组织化学（仅细胞周期蛋白D1）。 3）相关的染色体异常 肿瘤生物学标记（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非整倍性， 和浆膜形态学）。 4）评估特定染色体 与脊髓疾病和治疗相关的白血病和治疗相关的异常 将这些发现与TMDS/AML的临床发展有关。 材料和方法；为了检测染色体异常，他将执行 使用基因座特异性和染色体特异性探针在档案中使用双色鱼 250例E9487的患者的样本（相关的辅助实验室 临床试验试验E9486）。精确识别单型浆细胞 将对细胞质免疫球蛋白的鱼和荧光染色。 此外，他将对RT-PCR进行RNA分析和免疫组织化学的RT-PCR 蛋白质过表达（Cyclin D1）。他将执行生物相关性 使用其他标记，并执行整体生存和无事件生存 根据存在或不存在特定异常的分析。 还将使用多变量模型研究变量，以测试它们 独立的预测因子。