膜铁转运蛋白1在骨髓瘤发生、发展及骨质破坏中的作用及机制研究

Abnormal gene expression is shown to regulate the development and proliferation of multiple myeloma (MM) and evoke other pathological conditions such as bone destruction. But the role of ferroportin 1(FPN1) gene in the above-mentioned pathological process of MM is unclear at present. Our preliminary data have found that the FPN1 gene has lower expression in the vast majority of MM patients than those in normal people. Further, low expression of FPN1 is associated with inferior prognosis and serious bone destruction in MM patients. Over-expression of FPN1 significantly inhibits the growth of myeloma cells both in vivo and in vitro. We have developed FPN1 gene knockout mice which have an increase of the proliferation of plasma cells in bone marrow, bone marrow macrophage differentiates into osteoclast, and bone destruction. Based on our findings, we will investigate the role and mechanism of FPN1 in the development, progression, osteoclastogenesis, and bone destruction of MM by using gene chip analysis, western blot, flow cytometry analysis, two-dimensional electrophoresis of MS proteomics, CT, animal model etc. We will also determine the therapeutic efficacy of the FPN1 activator (sulforaphane, SFN) in combination with its downstream target inhibitors on myeloma disease progression and bone destruction. Furthermore, we will evaluate the relationship between FPN1 and myeloma prognosis and bone destruction. This study will provide new myeloma therapeutic targets and will also serve as a theoretical basis for the development of novel targeted agents to help clinical treatment of MM.

基因异常表达在多发性骨髓瘤（MM）发生、发展及骨质破坏等病理过程中起关键作用，但基因膜铁转运蛋白1（FPN1）在骨髓瘤上述病理过程中的作用目前尚不清楚。我们前期研究发现：大多数MM患者骨髓瘤细胞中FPN1低表达；FPN1低表达与MM患者预后差和严重骨破坏相关；MM细胞系过表达FPN1后，体内外MM细胞的生长都明显受到抑制；FPN1基因敲除小鼠骨髓中增殖的浆细胞显著增加、骨髓巨噬细胞分化为的破骨细胞明显增多、骨破坏加重。围绕上述发现，我们将应用基因芯片分析、Western blot、流式细胞仪分析、二维电泳-MS 蛋白质组学、CT、动物模型等方法进行研究，探讨FPN1在MM发生、发展和破骨细胞分化、骨破坏中的作用及机制；阐明莱菔硫烷等FPN1活化剂及下游信号靶向抑制剂对骨髓瘤生长及骨破坏的影响；分析FPN1与骨髓瘤预后及骨破坏的关系。本研究将为骨髓瘤提供新的治疗靶点，有助于临床治疗骨髓瘤。

基因异常表达在多发性骨髓瘤（MM）发生、发展及骨质破坏等病理过程中起关键作用，但膜铁转运蛋白1（FPN1）在MM上述病理过程中的作用目前尚不清楚。本项目采用基因表达谱分析、CRISPR-Cas9基因编辑、流式细胞仪检测、双荧光素酶报告实验、染色质免疫共沉淀技术和裸鼠皮下移植瘤动物模型等实验方法，并通过功能挽救实验和裸鼠移植瘤实验对体内和体外生物学行为进行验证。..我们研究发现：FPN1低表达在体外明显促进MM细胞的增殖和克隆形成，miR-17-5p可通过靶向结合FPN1的3’UTR区域而抑制FPN1的表达，核转录因子Nrf2可通过抑制miR-17-5p的转录进而调控FPN1的表达，Nrf2可直接结合FPN1的启动子区域转录调节其表达。此外，Nrf2/miR-17-5p轴可作为一个潜在的分子靶点共同调节机体活性氧和细胞内铁水平。本项目为MM治疗提供了新的治疗靶标，具有潜在的临床应用前景。同时，为了更深入研究MM发病机制，探讨干预MM的新策略，在本基金的资助下开展了具有潜在转化前景的新型小分子化合物抗MM作用的探索性研究，取得了可喜的成果。..已经发表标注本基金的SCI论文12篇，影响因子（IF）共60.53分，其中IF大于8分的2篇，IF在5-8分之间的5篇，中文期刊论文2篇。本基金第一标注的SCI论文9篇，IF共40.64分；本基金第二标注的SCI论文3篇，IF共19.89分；申请人是这些文章的通讯作者。申请国家发明专利5项（已获受理号，等待审批中），其中授权1项。培养博士研究生1名，硕士研究生1名。参加国内和国际学术交流11人次。.