摘要
目的利用DP7-C/DOTAP脂质体作为腺病毒运载工具,一方面通过DP7-C/DOTAP脂质体增强溶瘤腺病毒转染CAR受体低表达肿瘤细胞,克服溶瘤腺病毒在转染细胞时受到的CAR限制,提高转染效率;另一方面通过用DP7-C/DOTAP脂质体包裹溶瘤腺病毒,保护溶瘤腺病毒不被中和抗体清除。验证两方面增强溶瘤腺病毒的“溶瘤”作用。方法采用薄膜分散法制备DP7-C修饰的DOTAP脂质体(DP7-C/DOTAP),将腺病毒(Ad)与DP7-C/DOTAP脂质体在体外孵育形成DP7-C/DOTAP/Ad复合物。对DP7-C/DOTAP/Ad复合物的粒径、电位和形态学进行表征。然后在体外检测DP7-C/DOTAP/Ad复合物转染CAR受体低表达肿瘤细胞的效率及抵抗腺病毒中和抗体的功效。最后在小鼠中验证DP7-C/DOTAP脂质体促进溶瘤腺病毒H101的抗肿瘤效果。结果DP7-C/DOTAP脂质体其粒径分布在140~200 nm,电位为48~58 mV,透射电镜结果显示DP7-C/DOTAP/Ad复合物制备成功。与Ad相比,DP7-C/DOTAP/Ad复合物可显著提高低表达CAR受体卵巢癌细胞SKOV3的转染效率,同时DP7-C/DOTAP脂质体可以起到保护腺病毒不被中和抗体识别和清除的作用。在SKOV3卵巢癌皮下移植瘤模型中,单独H101组抑瘤率为23.90%,H101/DP7-C/DOTAP组抑瘤率为54.81%,可更好地抑制肿瘤生长(P<0.05)。结论DP7-C/DOTAP脂质体包裹腺病毒可增强腺病毒的转染效率和杀伤效果。DP7-C/DOTAP脂质体可有效促进腺病毒转染CAR低表达细胞。DP7-C/DOTAP脂质体可以对腺病毒起到良好的保护效果,不被抗腺病毒中和抗体中和。在动物实验中初步证明采用DP7-C/DOTAP脂质体能够增强溶瘤病毒的抗肿瘤效用。
Objective DP7-C/DOTAP liposomes were employed as carriers for oncolytic adenoviruses.On one hand,these liposomes enhanced the transfection of tumor cells with low CAR receptor expression,thereby overcoming the CAR-dependent entry limitation of oncolytic adenoviruses and improving transfection efficiency.On the other hand,by encapsulating oncolytic adenoviruses,DP7-C/DOTAP liposomes protected them from clearance by neutralizing antibodies.Together,these mechanisms enhanced the oncolytic effect of the adenovirus.Methods DP7-C-modified DOTAP liposomes(DP7-C/DOTAP)were prepared using the thin-film dispersion method.The complexes of adenovirus(Ad)and DP7-C/DOTAP liposomes(DP7-C/DOTAP/Ad)were formed via in vitro incubation.The particle size,zeta potential,and morphology of the complexes were characterized.Subsequently,the transfection efficiency of the DP7-C/DOTAP/Ad complex in CAR-low tumor cells and its ability to evade neutralization by anti-adenovirus antibodies were evaluated in vitro.Finally,the antitumor effect of DP7-C/DOTAP-encapsulated oncolytic adenovirus H101 was assessed in a mouse model.Results The DP7-C/DOTAP liposomes had a particle size of approximately 140-200 nm and a zeta potential of 48-58 mV.Transmission electron microscopy confirmed the successful preparation of the DP7-C/DOTAP/Ad complex.Compared with naked Ad,the DP7-C/DOTAP/Ad complex significantly enhanced the transfection efficiency in SKOV3 ovarian cancer cells with low CAR expression.Furthermore,the liposomes protected the adenovirus from recognition and clearance by neutralizing antibodies.In the SKOV3 xenograft model,the tumor inhibition rate was 23.90%for H101 alone and 54.81%for the H101/DP7-C/DOTAP group,indicating a significantly enhanced antitumor effect(P<0.05).Conclusions This study demonstrates that DP7-C/DOTAP liposomes enhance both the transfection efficiency and oncolytic effect of adenoviruses.They facilitate adenovirus entry into cells with low CAR expression and provide protection against neutralizing antibodies.In vivo,DP7-C/DOTAP liposomes significantly improved the antitumor efficacy of H101(P<0.05),preliminarily confirming their potential to enhance the therapeutic effect of oncolytic viruses.
作者
胡蝶
苍玉德
杨莉
HU Die;CANG Yude;YANG Li(Department of Biotherapy,Cancer Center and State Key Laboratory of Biotherapy,West China Hospital,Sichuan University,Chengdu 610041,China;Frontiers Medical Center,Tianfu Jincheng Laboratory,Chengdu 610212,China)
出处
《中国医药生物技术》
2025年第S2期1-11,共11页
Chinese Medicinal Biotechnology
基金
天府锦城实验室(前沿医学中心)团队赛马项目(TFJC2023010005)。
