Combined Antiviral Therapy Using Designed Molecular Scaffolds Targeting Two Distinct Viral Functions, HIV-1 Genome Integration and Capsid Assembly

• Type of Material: Article
• Authors: Wannisa Khamaikawin, Chiang Mai University; Somphot Saoin, Chiang Mai University; Sawitree Nangola, University of Phayao; Koollawat Chupradit, Chiang Mai University; Supachai Sakkhachornphop, Chiang Mai University; Sudarat Hadpech, Chiang Mai University; Nattawat Onlamoon, Mahidol University; Aftab Ansari, Emory University; Siddappa Byrareddy, Emory University; Pierre Boulanger, University Lyon 1 & INRA UMR-754; Saw-See Hong, University Lyon 1 & INRA UMR-754; Bruce E Torbett, The Scripps Research Institute; Chatchai Tayapiwatana, Chiang Mai University
• Language: English
• Date: 2015-08-25
• Publisher: Nature Publishing Group: Open Access Journals - Option B
• Publication Version: Final Published Version
• Copyright Statement: © 2015 Official journal of the American Society of Gene & Cell Therapy
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1038/mtna.2015.22
• Title of Journal or Parent Work: Molecular Therapy - Nucleic Acids
• ISSN: 2162-2531
• Volume: 4
• Start Page: e249
• End Page: e249
• Grant/Funding Information: Cluster and Program Management Office (CPMO); National Research University project under the Thailand's Office of the Commission on Higher Education; National Research Council of Thailand (NRCT); Health Systems Research Institute (HSRI); National Institutes of Health (NIH) grant CFAR-P30 AI036214, GM083658 and HL091219; Thailand Research Fund through the Royal Golden Jubilee Ph.D. program (grant number PHD/0024/2552 to W.K.); National Science and Technology Development Agency (NSTDA)
• Keywords: HIV-1-resistant cells; SHIV; designed zinc-finger proteins; anti-HIV-1 molecular scaffolds; SIV; designed ankyrin repeat proteins
• Research Categories: Biology, Virology; Health Sciences, Immunology

Abstract

Designed molecular scaffolds have been proposed as alternative therapeutic agents against HIV-1. The ankyrin repeat protein (Ank^GAG 1D4) and the zinc finger protein (2LTRZFP) have recently been characterized as intracellular antivirals, but these molecules, used individually, do not completely block HIV-1 replication and propagation. The capsid-binder Ank^GAG1D4, which inhibits HIV-1 assembly, does not prevent the genome integration of newly incoming viruses. 2LTRZFP, designed to target the 2-LTR-circle junction of HIV-1 cDNA and block HIV-1 integration, would have no antiviral effect on HIV-1-infected cells. However, simultaneous expression of these two molecules should combine the advantage of preventive and curative treatments. To test this hypothesis, the genes encoding the N-myristoylated Myr(+)Ank^GAG1D4 protein and the 2LTRZFP were introduced into human T-cells, using a third-generation lentiviral vector. SupT1 cells stably expressing 2LTRZFP alone or with Myr(+)Ank^GAG1D4 showed a complete resistance to HIV-1 in viral challenge. Administration of the Myr(+)Ank^GAG1D4 vector to HIV-1-preinfected SupT1 cells resulted in a significant antiviral effect. Resistance to viral infection was also observed in primary human CD4+ T-cells stably expressing Myr(+)Ank^GAG1D4, and challenged with HIV-1, SIVmac, or SHIV. Our data suggest that our two anti-HIV-1 molecular scaffold prototypes are promising antiviral agents for anti-HIV-1 gene therapy.