DELETION AND CHARACTERIZATION OF MYELOMA PRECURSORS

意义不明的单克隆丙种球蛋白病（MGUS）存在于3% 70岁以上的人。 我们的假设是， 多发性骨髓瘤（MM）和MGUS的特征在于存在 异常的骨髓（BM）浆细胞，它们在生物学上是不同的。 部分患者从MGUS向MM转化的机制尚不清楚 明白 从我们最初的研究中可以清楚地看出， 细胞可以在外周血（PB）中循环和增殖， 然而，他们仍然争论是否这种疾病 是在浆细胞水平上传播的，而不是前体B细胞。 这 建议将涉及两个实验室的合作， 来自MM和MGUS患者的PB和BM的分离样品的细胞， 更好地了解循环浆细胞的频率， 在这些疾病中发现了克隆相关的细胞， 这些细胞的分子特征。 MGUS患者的PBMNC 和MM将通过双色免疫荧光法分析浆细胞和前体B细胞。 免疫荧光显微镜、多参数DNA含量流式细胞术 (FC)三色免疫分型法。 等位基因特异 寡核苷酸（阿索）将用于敏感的和肿瘤特异性的 聚合酶链反应（PCR）技术在我们的实验室开发， 鉴定和定量恶性浆细胞和克隆相关细胞。 通过各种大小、免疫表型和DNA含量参数分选细胞 将用ASO-PCR技术进行检测，以了解特异性 恶性克隆的特征。 还将通过FC对细胞进行分选 转化为CD 38+和CD 38-组分，并通过逆转录PCR检测， 鉴定源自早期B细胞的克隆相关转录物 前体 我们的目标是学习1）做前体细胞或浆细胞 是否在MGUS患者的外周血中循环？ 2)克隆细胞可以 在MM或MGUS患者的pB中证实，当没有浆细胞时， 从而为循环前体细胞提供了证据？3)如果 前体细胞被发现，它们的分子和免疫表型是什么？和 4)单克隆浆细胞可以在基质细胞培养物中生长吗？ 来自分选细胞群的SCID小鼠？ 这些结果将更好地定义 PB细胞在产生和维持恶性细胞中的作用。 的 MGUS和MM患者的研究提供了一个更好的机会， 了解循环细胞的相似性和差异， 这两种疾病及其在疾病进展中的作用。

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.