title: Inhibition of BK virus replication in human kidney cells by BK virus large tumor antigen-specific shRNA delivered by JC virus-like particles.
abstract: Polyomavirus-associated nephropathy (PVAN) due to lytic infection by the BK polyomavirus (BKPyV) remains an important cause of allograft dysfunction and graft loss in renal transplant recipients. PVAN is commonly treated by reducing the dosage of immunosuppressive drugs and adding adjuvant antiviral agents, but the outcomes have been less than satisfactory. The BKPyV early protein large tumor antigen (LT) is indispensable for viral genome replication and viral late protein expression. Therefore, suppressing LT expression may be a way to inhibit BKPyV replication without harming the host human kidney cells. Previous studies have shown that JC polyomavirus (JCPyV) virus-like particles (VLPs), which have tropism for the human kidney, can package and transfer exogenous genes into human kidney cells for expression. In this study, we constructed an expression plasmid for a BKPyV LT-specific shRNA (shLT) and used JCPyV VLPs as a delivery vehicle to transduce the shLT plasmid into BKPyV-infected human kidney cells. The expression of BKPyV early (LT) and late (VP1) proteins was examined after transduction by immunofluorescence microscopy and Western blotting. We found that transduction with the shLT plasmid decreased the proportions of BKPyV LT- and VP1-expressing cells by 73% and 82%, respectively, relative to control. The viral genomes were also decreased by 56%. These results point to the promising possibility of developing shLT-transducing JCPyV VLPs as a specific anti-BKPyV approach for PVAN treatment.
authors.affiliation: Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan; Department of Urology, Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors.forename: Mien-Chun
authors.initials: MC
authors.lastname: Lin
authors.affiliation: Department of Microbiology and Immunology, Chung Shan Medical University, Taichung, Taiwan.
authors.forename: Meilin
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Medical Research, Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors.forename: Chiung-Yao
authors.initials: CY
authors.lastname: Fang
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan.
authors.forename: Pei-Lain
authors.initials: PL
authors.lastname: Chen
authors.affiliation: Department of Urology, Chia-Yi Christian Hospital, Chia-Yi, Taiwan. Electronic address: 01712@cych.org.tw.
authors.forename: Cheng-Huang
authors.initials: CH
authors.lastname: Shen
authors.affiliation: Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan. Electronic address: biodcc@ccu.edu.tw.
authors.forename: Deching
authors.initials: D
authors.lastname: Chang
mesh_terms: D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism*; D000998:Antiviral Agents / Q000378:metabolism*; D001739:BK Virus / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D007577:JC Virus / Q000235:genetics; D010957:Plasmids; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D014764:Viral Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000956:Antigens, Viral; D000998:Antiviral Agents; D034741:RNA, Small Interfering; D014764:Viral Proteins
keywords: BK polyomavirus; BKPyV replication; Gene therapy; JCPyV VLP; Short hairpin RNA