聚合物-脂质纳米粒用于增强酪氨酸血症I型治疗性碱基编辑器质粒的肝靶向递送研究

Polymer-lipid nanoparticles enhance liver-targeted delivery of therapeutic base editor plasmid for the treatment of hereditary tyrosinemia type 1(HT-1)

酪氨酸血症I型是一种罕见的常染色体隐性遗传病,目前尚无有效的治疗方法。近年来,以碱基编辑器为代表的基因编辑技术已被报道用于酪氨酸血症I型的治疗。然而,由于生理屏障的存在,碱基编辑器递送困难。在本研究中,我们构建了一种靶向去唾液酸糖蛋白受体的聚合物-脂质纳米递送系统,用于改善酪氨酸血症I型治疗性核酸药物的递送效率。我们首先合成了一种生物可降解性丙烯酸酯-氨基醇共聚物用于递送碱基编辑器质粒,其转染效率显著优于市售转染试剂Hieff Trans^(TM)。随后,共聚物纳米粒与DOPE-PEG-Gal NAc自组装形成聚合物-脂质纳米粒,用于增强纳米粒的肝脏递送效率。在体外转染实验中,包载Fah-p CMV-ABE6.3-EGFP碱基编辑器质粒的聚合物-脂质纳米粒表现出了良好的肝细胞选择性,其转染效率是游离质粒的70倍以上。研究表明,携带肝靶向配体的聚合物-脂质纳米递送系统能够有效增强治疗性碱基编辑器质粒的肝靶向递送效率并为酪氨酸血症I型的基因治疗提供了一种潜在的递送载体。

Hereditary tyrosinemia type 1(HT-1)is a rare autosomal recessive genetic disease with no effective cure at presen t.In recent years,gene-editing techniques such as base editor(BE)have been explored for treating HT-1.However,the delivery of nucleic acids faces challenges due to existing physiological barriers.In the present study,we constructed an asialoglycoprotein receptor(ASGPR)-targeted polymer-lipid nanoparticle to enhance the delivery efficiency of therapeutic nucleic acids for HT-1.To deliver BE plasmids,a biodegradable cationic polymer,acrylate-amino alcohol copolymer was synthesized and demonstrated superior transfection efficiency compared to the commercially available transfection reagent,Hieff TransTM.Subsequently,DOPE-PEG-GalNAc was combined with copolymer nanoparticles to enhance the hepatocyte delivery of the nanoparticles.Upon loading the recombinant BE plasmid,Fah-pCMV-ABE6.3-EGFP,the polymer-lipid nanoparticles exhibited remarkable hepatocyte selectivity,with a transfection efficiency over 70-fold higher than that of the free plasmid.The study suggested that polymer-lipid nanoparticles,in combination with liver-targeted ligands,could effectively enhance the liver-targeted delivery of therapeutic BE plasmids,providing a promising vector for gene therapy of HT-1.