Anti EGFR scFv::FTH1/FTH1纳米粒子的制备及其在哮喘治疗中的应用

Preparation of anti EGFR scFv::FTH1/FTH1 nanoparticles for asthma treatment

摘要:

哮喘作为常见的呼吸性疾病之一，严重危害人类的健康，造成了严重的医疗负担。最新研究表明表皮生长因子受体 (Epidermal growth factor receptor，EGFR) 参与哮喘的发生发展。为构建靶向EGFR用于哮喘治疗的纳米粒子，通过基因工程方法将抗EGFR的单链抗体 (Single chain antibody fragment，scFv) 修饰在铁蛋白重链亚基 (Human ferritin H-chain，FTH1) N端，采用混合复性的方式成功构建anti EGFR scFv::FTH1/FTH1纳米粒子。通过透射电子显微镜对纳米粒子的结构进行分析，结果表明，纳米粒子可以自组装成中空笼状结构，粒径约为12 nm。采用SDS-PAGE对纯化后的纳米粒子进行半定量分析发现，亚基物质的量之比为FTH1︰anti EGFR scFv::FTH1=7︰3。在哮喘小鼠动物模型上发现Anti EGFR scFv::FTH1/FTH1纳米粒子能有效抑制哮喘小鼠肺组织中杯状细胞增生及粘液分泌，对胶原沉积纤维化也有一定抑制效果。这些研究为铁蛋白应用于哮喘的治疗提供了良好的基础。

Abstract:

Asthma is a common respiratory disease that affects 300 million of people worldwide, posing a serious health risk and medical burden. Development of new anti-asthmatic drugs and alternative treatment regimens is therefore encouraged. Recent studies have shown that Epidermal Growth Factor Receptor (EGFR) is involved in asthma development. In order to construct nanoparticles targeting EGFR for asthma treatment, a single chain antibody fragment (scFv) against EGFR was genetically engineered and modified at the N-terminal end of the human ferritin H-chain (FTH1) to construct Anti EGFR scFv::FTH1/FTH1 nanoparticles. Transmission electron microscopy showed that the nanoparticles were self-assembled into hollow cage-like structures with the particle size of about 12 nm. Semi-quantitative analysis of the purified nanoparticles by SDS-PAGE revealed the mass ratio of FTH1 to Anti EGFR scFv::FTH1 was 7:3. In House Dust Mite (HDM) driven models, Anti EGFR scFv::FTH1/FTH1 nanoparticles efficiently attenuated several key features of asthma, including goblet cell hyperplasia, mucous metaplasia and subepithelial fibrosis, showing the potential of using ferritin based nanoparticle for asthma treatment.