From the Department of Pathology and Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama 35294 and.
J Biol Chem. 2014 Feb 28;289(9):5499-509. doi: 10.1074/jbc.M113.532143. Epub 2014 Jan 8.
Recently, it was discovered that serglycin, a hematopoietic cell proteoglycan, is the major proteoglycan expressed and constitutively secreted by multiple myeloma (MM) cells. High levels of serglycin are present in the bone marrow aspirates of at least 30% of newly diagnosed MM patients. However, its contribution to the pathophysiology of MM is unknown. Here, we show that serglycin knockdown (by ∼85% compared with normal levels), using lentiviral shRNA, dramatically attenuated MM tumor growth in mice with severe combined immunodeficiency. Tumors formed from cells deficient in serglycin exhibited diminished levels of hepatocyte growth factor expression and impaired development of blood vessels, indicating that serglycin may affect tumor angiogenesis. Furthermore, knockdown of serglycin significantly decreased MM cell adhesion to bone marrow stromal cells and collagen I. Even though serglycin proteoglycan does not have a transmembrane domain, flow cytometry showed that serglycin is present on the MM cell surface, and attachment to the cell surface is, at least in part, dependent on its chondroitin sulfate side chains. Co-precipitation of serglycin from conditioned medium of MM cells using a CD44-Fc chimera suggests that CD44 is the cell surface-binding partner for serglycin, which therefore may serve as a major ligand for CD44 at various stages during myeloma progression. Finally, we demonstrate that serglycin mRNA expression in MM cells is up-regulated by activin, a predominant cytokine among those increased in MM patients with osteolytic lesions. These studies provide direct evidence for a critical role for serglycin in MM pathogenesis and show that targeting serglycin may provide a novel therapeutic approach for MM.
最近发现,一种造血细胞蛋白聚糖——硫酸乙酰肝素蛋白聚糖(Serglycin)是多发性骨髓瘤(MM)细胞表达和持续分泌的主要蛋白聚糖。在至少 30%的新诊断 MM 患者的骨髓抽吸物中存在高水平的硫酸乙酰肝素蛋白聚糖。然而,其对 MM 病理生理学的贡献尚不清楚。在这里,我们展示了使用慢病毒 shRNA 敲低(与正常水平相比降低约 85%)硫酸乙酰肝素蛋白聚糖,可显著抑制严重联合免疫缺陷小鼠中的 MM 肿瘤生长。来自缺乏硫酸乙酰肝素蛋白聚糖的细胞形成的肿瘤表现出肝细胞生长因子表达水平降低和血管发育受损,表明硫酸乙酰肝素蛋白聚糖可能影响肿瘤血管生成。此外,敲低硫酸乙酰肝素蛋白聚糖可显著降低 MM 细胞与骨髓基质细胞和胶原 I 的黏附。尽管硫酸乙酰肝素蛋白聚糖没有跨膜结构域,但流式细胞术显示硫酸乙酰肝素蛋白聚糖存在于 MM 细胞表面,并且与细胞表面的附着至少部分依赖于其软骨素硫酸盐侧链。使用 CD44-Fc 嵌合体从 MM 细胞的条件培养基中沉淀硫酸乙酰肝素蛋白聚糖表明,CD44 是硫酸乙酰肝素蛋白聚糖的细胞表面结合伴侣,因此它可能在骨髓瘤进展的各个阶段作为 CD44 的主要配体。最后,我们证明激活素(在伴有溶骨性病变的 MM 患者中增加的主要细胞因子之一)可上调 MM 细胞中的硫酸乙酰肝素蛋白聚糖 mRNA 表达。这些研究为硫酸乙酰肝素蛋白聚糖在 MM 发病机制中的关键作用提供了直接证据,并表明靶向硫酸乙酰肝素蛋白聚糖可能为 MM 提供一种新的治疗方法。