title: Development of next generation adeno-associated viral vectors capable of selective tropism and efficient gene delivery.
abstract: Virus-based nanoparticles have shown promise as vehicles for delivering therapeutic genes. However, the rational design of viral vectors that enable selective tropism towards particular types of cells and tissues remains challenging. Here, we explored structural-functional relationships of the adeno-associated virus 2 (AAV2) vector by expanding its genetic code during production. As a proof-of-principle, an azide moiety was strategically displayed on the vector capsid as a bioorthogonal chemical reporter. Upon bioorthogonal conjugation of AAV2 with fluorophores and cyclic arginyl-glycyl-aspartic acid ligands at certain modifiable sites, we characterized in vitro and in vivo AAV2 movement and enhanced tropism selectivity towards integrin-expressing tumor cells. Targeting AAV2 vectors resulted in selective killing of U87 glioblastoma cells and derived xenografts via the herpes simplex virus suicide gene thymidine kinase, with the potency of ganciclovir being increased by 25-fold. Our results demonstrated successful rational modification of AAV2 as a targeting delivery vehicle, establishing a facile platform for precision engineering of virus-based nanoparticles in basic research and therapeutic applications.
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Chuanling
authors.initials: C
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Tianzhuo
authors.initials: T
authors.lastname: Yao
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Yongxiang
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Zhongjun
authors.initials: Z
authors.lastname: Li
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China.
authors.forename: Lihe
authors.initials: L
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, 100191, China. Electronic address: deminzhou@bjmu.edu.cn.
authors.forename: Demin
authors.initials: D
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D036022:Capsid Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D002460:Cell Line; D045744:Cell Line, Tumor; D000229:Dependovirus / Q000737:chemistry / Q000235:genetics*; D004337:Drug Carriers / Q000737:chemistry; D018014:Gene Transfer Techniques; D005815:Genetic Code; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D005909:Glioblastoma / Q000235:genetics / Q000628:therapy; D006801:Humans; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry; D015202:Protein Engineering; D014161:Transduction, Genetic
chemical_list: D036022:Capsid Proteins; D004337:Drug Carriers; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid
keywords: Adeno-associated virus; Genetic code expansion; Structure-function relationships; Targeted delivery; Unnatural amino acids