Melittin analog p5RHH enhances recombinant adeno-associated virus transduction efficiency  

作　　者：Jing-shun Meng Yun He Heng-bin Yang Li-ping Zhou Si-yuan Wang Xi-lin Feng Omar Yahya Al-shargi Xiao-min Yu Li-qing Zhu Chang-quan Ling 

机构地区：[1]Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China [2]Oncology Department of Traditional Chinese Medicine,Changhai Hospital,Naval Medical University,Shanghai 200433,China [3]Faculty of Traditional Chinese Medicine,Naval Medical University,Shanghai 200043,China [4]State Key Laboratory of Genetic Engineering and Engineering Research Center of Gene Technology(Ministry of Education),School of Life Sciences,Zhongshan Hospital,Fudan University,Shanghai 200438,China [5]Yantai Fuheng Biological Technology Co.,Ltd.,Yantai,Shandong Province 264006,China. [6]Department of Clinical Laboratory,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,Zhejiang Province,China

出　　处：《Journal of Integrative Medicine》2024年第1期72-82,共11页结合医学学报（英文版）

基　　金：sponsored by grants from the National Natural Science Foundation of China(No.82030117);the Wenzhou major scientific and technological innovation project(No.ZY2022001).

摘　　要：Objective Melittin and its derivatives have been characterized to establish effective gene delivery systems.Their capability of facilitating endosomal release enhances the nanoparticles-based gene delivery.Nevertheless,little investigation has been conducted to explore its potential application in the context of viral vectors.Methods Various melittin-derived peptides were inserted into the loop VIII of the capsid proteins of recombinant adeno-associated virus vectors.These vectors carrying either gfp or fluc genes were subjected to qPCR assays and transduction assays of HEK293T cells to investigate the efficiency of vector production and gene delivery.In addition,the ability of a specific p5RHH-rAAV vector to deliver genes was examined through in vitro transduction of different cultured cells and in vivo tail vein administration to C57BL/6 mice.Finally,the intricate details of the vector-mediated transduction mechanisms were revealed by specific pharmacological inhibitors of every stage of the rAAV2 intracellular life cycle.Results A total of 76 melittin-related peptides were compiled from existing literature.Among them,cMA2,Melt13,p5RHH and aAR3 were found to significantly enhance the gene delivery efficiency of rAAV2 vectors.The p5RHH-rAAV2 vectors efficiently transduced not only rAAV-potent cell lines but also cell lines previously considered resistant to rAAV.Mechanistically,bafilomycin A1,a vacuolar endosome acidification inhibitor,completely inhibited the transgene expression mediated by the p5RHH-rAAV2 vectors.Most importantly,p5RHH-rAAV8 vectors also demonstrated increased hepatic transduction in vivo in C57BL/6 mice.Conclusion The incorporation of melittin analogues into the rAAV capsids results in a significant improvement in rAAV-mediated transgene expression.While further modifications remain an area of interest,our studies have substantially broadened the pharmacological prospects of melittin in the context of viral vector-mediated gene delivery.

关 键 词：MELITTIN Recombinant adeno-associated virus Capsid engineering Transduction efficiency 

分 类 号：Q78[生物学—分子生物学]