DELETION AND CHARACTERIZATION OF MYELOMA PRECURSORS

未确定意义的单克隆性伽马病(MGUS)出现在3% 年龄超过70岁的人。我们的假设是尽管两者 多发性骨髓瘤(MM)和MGUS的特征是 异常的骨髓(BM)浆细胞，它们在生物学上是不同的。 一些患者从MGUS转化为MM的机制不佳 明白了。从我们的初步研究中可以清楚地看到，异常血浆 人外周血中的细胞可以循环和增殖 然而，患有MM的患者，他们的遗体对这种疾病是否 是在浆细胞水平上繁殖的，而不是前体B细胞。这 提案将涉及两个实验室的合作，以检查 MM和MGUS患者外周血和骨髓分离样本中的细胞 更好地了解浆细胞和 在这些疾病中发现克隆相关细胞，免疫学和 这些细胞的分子特征。MGUS患者的外周血单核细胞 MM的浆细胞和前体B细胞将进行双色分析 免疫荧光显微镜、多参数DNA含量流式细胞术 (Fc)，Fc三色免疫表型。一种等位基因特异性 寡核苷酸(ASO)将用于一种敏感的和肿瘤特异性的 本实验室发展的聚合酶链式反应(PCR)技术 鉴定和定量恶性浆细胞和克隆性相关细胞。 根据不同大小、免疫表型和DNA含量参数对细胞进行分类 将通过ASO-PCR技术进行检查，以了解特定的 恶性克隆的特征。单元格还将按本币进行排序 转化为CD38+和CD38-组分，用逆转录聚合酶链式反应检测 识别来自早期B细胞的克隆相关转录本 先驱物。我们的目标是学习1)前体细胞或浆细胞 在MGUS患者外周血中循环？2)克隆性细胞 MM或MGUS患者外周血中无浆细胞 从而为循环中的前体细胞提供了证据？3)如果 发现前体细胞，它们的分子和免疫表型是什么？和 4)单克隆性浆细胞可以在基质细胞培养中生长，还是在 从分类的细胞群中分离的SCID小鼠？这些结果将更好地定义 外周血细胞在产生和维持恶性细胞中的作用。这个 对MGUS和MM患者的研究提供了一个更好的机会 了解中发现的循环细胞的异同 这两种疾病及其在疾病发展中的作用。

Monoclonal gammopathy of undetermined significance (MGUS) is present in 3% of people more 70 years of ag e. It is our hypothesis that although both multiple myeloma (MM) and MGUS are characterized by the presence of abnormal bone marrow (BM) plasma cells, they are biologically different. The mechanisms of why some patients transform from MGUS to MM are poorly understood. It is clear from our initial studies that abnormal plasma cells can circulate and proliferate in the peripheral blood (PB) of patients with MM. However, their remains debate as to whether the disease is propagated at the plasma cell level versus a precursor B-cell. This proposal will involve the collaboration of two laboratories to examine cells from split-samples of the PB and BM of patients with MM and MGUS to gain a better understanding of how frequently circulating plasma cells and clonally-related cells are found in these diseases and the immunologic and molecular characteristics of these cells. PBMNC from patients with MGUS and MM will be analyzed for plasma cells and precursor B-cells by two-color immunofluorescence microscopy, multiparameter DNA content flow cytometry (FC), and three-color immunophenotyping by FC. An allele-specific oligonucleotide (ASO) will be used in a sensitive and tumor-specific polymerase chain reaction (PCR) technique developed in our laboratory to identify and quantitate malignant plasma cells and clonally-related cells. Cells sorted by various size, immunophenotypic, and DNA content parameters will be examined with the ASO-PCR technique to learn the specific characteristics of the malignant clone. Cells will also be sorted by FC into CD38+ and CD38- fractions and examined by reverse transcript PCR to identify clonally related transcripts that are derived from early B-cell precursors. We aim to learn 1) Do precursor cells or plasma cells circulate in the pB of patients with MGUS? 2) Can clonal cells be demonstrated in the pB of patients with MM or MGUS when no plasma cells are found thus providing evidence for circulating precursor cells? 3) If precursor cells are found, what is their molecular and immunophenotype? and 4) Can monoclonal plasma cells be grown in stromal cell cultures or in SCID mice from sorted cell populations? These results will better define the role of the PB cells in producing and maintaining malignant cells. The study of both MGUS and MM patients provides an opportunity to better understand the similarities and differences in circulating cells found in these two diseases and their role in disease progression.