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Development of Organ-specific Treatment for Bone Metastasis in Neuroblastoma

神经母细胞瘤骨转移器官特异性治疗的进展

基本信息

批准号：
14570793
负责人：
MICHIGAMI Toshimi
金额：
$ 1.86万
依托单位：
Osaka Medical Center and Research Institute for Maternal and Child Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570793/
关键词：
neuroblastoma bone metastasis RANKL gene therapy osteoprotegerin 破骨細胞筋肉内遺伝子導入

项目摘要

Bone metastasis in neuroblastoma is an unfavorable prognostic factor even with intensive therapy. We have previously reported that nuclear factor κB ligand (RANKL) expression is stimulated in the coculture of human neuroblastoma cell line and murine bone marrow cells containing osteoclast precursors and stromal cells, compared with the culture of bone marrow cells alone. As the result, osteoclastogenesis was increased in the coculture. In other cancers such as prostate cancer and multiple myeloma as well, RANKL signaling is suggested to play a role. In an attempt to develop an organ-specific treatment for bone metastasis in neuroblastoma, we examined the effect of gene therapy using osteoprotegerin (OPG) expression vector in some animal models of osteolytic bone diseases. As the expression vector, we utilized pCAGGS containing CAG promoter. We constructed pCAGGS-OPG-FLAG, and confirmed the biological activity of OPG-FLAG in an in vitro osteoclastogenesis assay. To introduce pCAGGS-OPG-FLAG into the muscle of animals, we utilized naked DNA injection method combined with electroporation. In animal models of humoral hypercalcemia of malignancy, introduction of pCAGGS-OPG-FLAG restored hypercalcemia. In OPG knockout mice, the treatment using pCAGGS-OPG-FLAG increased bone mineral density. These data suggested the usefulness of treatment targeting RANKL signaling in osteolytic bone diseases, containing bone metasatsis in neuroblastoma.

神经母细胞瘤的骨转移是一个不利的预后因素，即使经过强化治疗。我们以前曾报道，与单独培养骨髓细胞相比，在人神经母细胞瘤细胞系和含有破骨细胞前体和基质细胞的小鼠骨髓细胞的共培养物中，核因子κB配体（RANKL）的表达受到刺激。其结果是，破骨细胞生成增加的共培养。在其他癌症如前列腺癌和多发性骨髓瘤中，RANKL信号传导也被认为发挥作用。为了开发一种针对神经母细胞瘤骨转移的器官特异性治疗方法，我们在一些溶骨性骨病动物模型中检测了使用骨保护素（OPG）表达载体的基因治疗效果。作为表达载体，我们利用含有CAG启动子的pCAGGS。我们构建了pCAGGS-OPG-FLAG，并在体外破骨细胞生成实验中证实了OPG-FLAG的生物学活性。为了将pCAGGS-OPG-FLAG导入动物肌肉中，我们采用裸DNA注射法结合电穿孔法。在恶性肿瘤的体液性高钙血症的动物模型中，引入pCAGGS-OPG-FLAG恢复了高钙血症。在OPG敲除小鼠中，使用pCAGGS-OPG-FLAG的治疗增加了骨矿物质密度。这些数据表明靶向RANKL信号传导的治疗在溶骨性骨病中的有用性，包括神经母细胞瘤中的骨转移。