构建小鼠Graves'病模型及靶向ICAM-1和TSHR治疗的策略优化研究

The Graves’ disease (GD) animal model building has not been widely agreed upon by academics due to the complex model preparation process, low success rate and poor repeatability. The establishment of GD animal model contributes to the study of aetiological agent and therapeutic method of it. Our published results confirmed that GD mice model was built successfully by intramuscular injecting recombinant plasmid （pcDNA3.1/TSHR268） and EP at the same time and on the same location to enhance immunization. The new method has advantages of short modeling cycle, high positive rate, good stability and repeatability. In other previous researches, ICAM-1 was found to play a role in occurrence and development of GD. And as an autoantigen of GD, thyroid stimulating hormone receptor (TSHR) plays a key role in onset and progression of GD. On this basis, we plan to use the anti ICAM-1 mAb with biological activity and the siRNA with inhibition target of TSHR for targeting therapy research of model mice’s GD. We will compare the results of above treatment with radionuclide therapy and study the possibility of anti ICAM-1 mAb and siRNA for GD treatment. Based on the above research, we can do further study on etiology and pathogenesis of GD, and provide new ideas for clinical treatment of it.

Graves’病（GD）动物模型制备过程复杂、成功率低、可重复性差，目前缺乏公认构建方法。建立符合临床GD的动物模型，对于研究其发病机理，以及开发新的治疗策略具有重要意义。我们前期已公开发表的研究证实，使用重组质粒（pcDNA3.1/TSHR268）肌肉注射免疫小鼠，并在注射部位电穿孔，可成功构建小鼠GD模型，该方法建模周期短、阳性率高、稳定性及可重复性好。基于现有研究，ICAM-1与GD的发生与发展密切相关，而TSHR是GD的自身抗原，在GD的发病及病情进展中至关重要。本项目拟采用靶向ICAM-1及TSHR的方法，构建具有生物活性的抗ICAM-1单抗和以TSHR为抑制靶点的siRNA，对模型小鼠的GD进行实验性治疗，通过与传统的放射性核素治疗比较，研究抗ICAM-1单抗和siRNA治疗GD的可能性，从而对GD的病因、发病机制进行深入研究，为GD的临床治疗提供新思路。

格雷夫斯病(Graves’ disease，GD) 是一种器官特异性自身免疫性甲状腺疾病，目前病因尚未完全阐明，同时也是甲亢最常见的病例类型，约占80%以上。目前临床治疗GD的方法主要有三种：抗甲状腺药物治疗、放射性碘及外科手术治疗。三种疗法均可不同程度控制GD病情，但均不能针对病因治愈GD。构建GD动物模型有助于对GD的病因、发病机制及治疗方法进行研究。本研究成功构建了制作周期短、阳性率高、稳定性及可重复性好的BALB/c小鼠GD模型。细胞间黏附分子-1（intercellular adhesion molecule-1, ICAM-1）与GD的发生与发展密切相关，而促甲状腺激素受体（thyroid stimulating hormone receptor, TSHR）是GD的自身抗原，在GD的发病及病情进展中至关重要。本研究构建具有生物活性的抗ICAM-1单克隆抗体（anti ICAM-1 mAb）和以TSHR为抑制靶点的小干扰RNA（siRNA），对模型小鼠的GD进行实验性治疗。两种治疗方法均可使GD模型小鼠的促甲状腺激素（thyroid-stimulating hormone, TSH）升高，甲状腺素（thyroxine, T4）、甲状腺刺激性抗体（thyroid stimulating antibody, TSAb）、甲状腺刺激阻断性抗体（thyroid stimulation-blocking antibody, TSBAb）不同程度降低。同时治疗后的模型小鼠甲状腺摄取能力降低，从而取得满意的治疗效果。但是不同方法治疗后模型小鼠甲状腺病理学检查与未治疗的小鼠相比，未见明显差异。此外由于抗ICAM-1单抗组的血清学指标改善更为明显，其治疗效果优于siRNA组。本研究以成功构建BALB/c小鼠GD模型为基础，探索GD治疗的新方法，对GD的病因、发病机制进行深入研究，为GD的临床治疗提供新思路。