title: Specific transgene expression in HIV-infected cells using protease-cleavable transcription regulator.
abstract: Gene therapy is a promising strategy for the treatment of HIV infection, but cell specificity remains an issue. Recently we have developed a new concept for a drug or gene delivery system responding to cellular signals (D-RECS) to achieve cell-specific transgene expression using a non-viral polymer-based vehicle. According to this concept, intracellular signaling enzymes, which are activated specifically in target cells, are used to trigger transgene expression. We previously applied this concept to HIV-1 protease and showed that the recombinant protease could act as a suitable signal. Here we further developed this system to achieve highly specific transgene expression in HIV-infected cells. We prepared a polymeric gene regulator grafted with a cationic peptide containing the HIV-Tat peptide via a specific substrate for HIV-1 protease. The regulator formed a stable polyplex with the transgene, suppressing its transcription. HIV-1 protease cleaved the peptide and released the transgene, which was consequently expressed specifically in activated HIV-infected cells, but remained unreleased and inactive in uninfected cells. The validity of this approach was further confirmed by applying it to the CVB1 2A protease of coxsackievirus (Picornaviridae family). This strategy should be widely applicable for specific expression of a variety of therapeutic genes in virus-infected cells.
authors.affiliation: Department of Microbiology, St. Marianna University School of Medicine, Kawasaki 216-8511, Japan. asai@marianna-u.ac.jp
authors.forename: Daisuke
authors.initials: D
authors.lastname: Asai
authors.forename: Masanori
authors.initials: M
authors.lastname: Kuramoto
authors.forename: Yoko
authors.initials: Y
authors.lastname: Shoji
authors.forename: Jeong-Hun
authors.initials: JH
authors.lastname: Kang
authors.forename: Kota Bae
authors.initials: KB
authors.lastname: Kodama
authors.forename: Kenji
authors.initials: K
authors.lastname: Kawamura
authors.forename: Takeshi
authors.initials: T
authors.lastname: Mori
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Miyoshi
authors.forename: Takuro
authors.initials: T
authors.lastname: Niidome
authors.forename: Hideki
authors.initials: H
authors.lastname: Nakashima
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Katayama
mesh_terms: D002412:Cations / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry*; D015696:Gene Products, tat / Q000737:chemistry / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D015658:HIV Infections / Q000628:therapy; D016333:HIV Protease / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D019169:Jurkat Cells; D008156:Luciferases / Q000235:genetics; D010316:Particle Size; D010455:Peptides / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity; D014158:Transcription, Genetic; D019076:Transgenes
chemical_list: D002412:Cations; D004337:Drug Carriers; D015696:Gene Products, tat; D010455:Peptides; D011108:Polymers; D004247:DNA; D008156:Luciferases; D016333:HIV Protease; C514803:p16 protease, Human immunodeficiency virus 1