Melittin analog p5RHH enhances recombinant adeno-associated virus transduction efficiency

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外文摘要：Objective:Melittin and its derivative have been developed to support effective gene delivery systems.Their ability to facilitate endosomal release enhances the delivery of nanoparticle-based gene therapy.Nevertheless,its potential application in the context of viral vectors has not received much attention.Therefore,we would like to optimize the rAAV vector by Melittin analog to improve the transduction effi-ciency of rAAV in liver cancer cells and explore the mechanism of Melittin analog on rAAV.Methods:Various melittin-derived peptides were inserted into loop Ⅷ of the capsid protein in recom-binant adeno-associated virus vectors.These vectors carrying either gfp or fluc genes were subjected to quantitative polymerase chain reaction assays and transduction assays in human embryonic kidney 293(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 explored by using pharmacological inhi-bitors of every stage of the rAAV2 intracellular life cycle.Results:A total of 76 melittin-related peptides were identified from existing literature.Among them,CMA-3,p5RHH and aAR3 were found to significantly inhibit transduction of rAAV2 vector crude lysate.The p5RHH-rAAV2 vectors efficiently transduced not only rAAV-potent cell lines but also cell lines pre-viously 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 increased hepatic transduction in vivo in C57BL/6 mice.Conclusion:The incorporation of melittin analogs into the rAAV capsids results in a significant improve-ment 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.

外文关键词：

MelittinRecombinant adeno-associated virusCapsid engineeringTransduction 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

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作者单位：

Faculty of Traditional Chinese Medicine,Naval Medical University,Shanghai 200043,China

Oncology Department of Traditional Chinese Medicine,Changhai Hospital,Naval Medical University,Shanghai 200433,China

Department of Oncology,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China

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

Yantai Fuheng Biological Technology Co.,Ltd.,Yantai 264006,Shandong Province,China

Department of Clinical Laboratory,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,Zhejiang Province,China

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基金：

国家自然科学基金Wenzhou major scientific and technological innovation project

项目编号：

82030117ZY2022001

出版年：

2024

DOI：

10.1016/j.joim.2024.01.001

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