MOLECULAR CYTOGENETICS OF MULTIPLE MYELOMA

多发性骨髓瘤(MM)是一种致命性肿瘤，其发生、发展和临床行为的遗传机制尚不清楚。建议的研究基于这样一种假设，即利用当前可用的尖端技术对MM的详细遗传特征将使人们能够对这种重要的血液系统恶性肿瘤的生物学和临床行为有新的理解。在这些研究中，我们建议将光谱核型分析(SKY)和比较基因组杂交(CGH)这两种新的分子细胞遗传学技术与染色体易位失控基因的克隆和特征相结合。这项应用有三个具体目标：1.使用CGH和SKY确定MM的染色体变化我们将把分子细胞遗传学技术SKY和CGH应用于一系列预期确定的MM肿瘤样本，以进行诊断评估并进入纪念医院(MH)治疗方案，以便全面描述疾病所有阶段的结构重排(Sky)和数值异常(CGH)。2.分离与IG基因重排失控的新基因。我们将从新的与14q32(IgH)相关的染色体易位中分离和鉴定候选的去调控基因。我们在第一次天空研究中发现的两个易位t(12；14)(q24；q32)和t(14；20)(q32；q11)将被初步定位于克隆。3.开发一种新的MM的遗传预后模型。我们将利用Sky和CGH关于染色体区域的反复重排、断裂点和得失的数据，以及传统的预后标记物，如β2-微球蛋白、血清IL-2-6和C-反应蛋白的水平，来开发MM的遗传预后模型。CGH确定的次区域得失也可能确定其扩增或失活可能影响临床结果的基因的位置。我们对多发性骨髓瘤的初步研究以及我们之前在B细胞非霍奇金淋巴瘤(NHL)分析中取得的类似目标的成功，给了我们信心，我们将在本申请中提出的研究中取得成功。

Multiple myeloma (MM) is a fatal neoplastic disease whose genetic mechanisms of origin, progression, and clinical behavior are poorly understood. The proposed studies are based on the hypothesis that detailed genetic characterization of MM utilizing currently-available cutting-edge technologies will enable a new understanding of the biology and clinical behavior of this important hematopoietic malignancy. In these studies, we propose to combine the two novel molecular cytogenetic techniques, spectral karyotyping (SKY) and comparative genomic hybridization (CGH), with those of cloning and characterization of genes deregulated in chromosomal translocations. This applicatin has three Specific Aims: 1. Define chromosomal changes in MM using CGH and SKY We will apply the molecular cytogenetic techniques SKY and CGH to a series of prospectively ascertained MM tumor samples seen for diagnostic evaluation and entered on Memorial Hospital (MH treatment protocols in order to fully characterize structural rearrangements (SKY) and numerical abnormalities (CGH) at all stages of the disease. 2. Isolate novel genes deregulated by rearrangement with IG genes. We will isolate and characterize the candidate deregulated genes from novel 14q32 (IGH)-associated chromosomal translocations. The two translocations, t(12;14)(q24;q32) and t(14;20)(q32;q11), discovered in the first SKY study of these tumors by us will be initially targeted for cloning. 3. Develop a new genetic prognostic model for MM. We will develop a genetic prognostic model for MM utilizing data from SKY and CGH on recurring rearrangements, breakpoints, and gains and losses of chromosomal regions, along with conventional prognostic markers such as levels of beta2-microglobulin, serum IL-2-6, and C-reactive protein, on patients entered on Memorial Hospital treatment protocols. The subregional gains and losses identified by CGH may also identify sites of genes whose amplification or inactivation may influence clinical outcome. Our preliminary studies of MM and our previous success with similar goals in the analysis of B-cell non-Hodgkin's lymnphoma (NHL), give us the confidence that we will be successful in the studies proposed in this application.