Engineered Zinc Finger Protein Targeting 2LTR Inhibits HIV Integration in Hematopoietic Stem and Progenitor Cell-Derived Macrophages: In Vitro Study

International Journal of Molecular Sciences

Chatchai Tayapiwatana¹  Chaniporn Puaninta²  Suparerk Borwornpinyo³  Bruce E. Torbett⁴  Suradej Hongeng⁵  Wannisa Khamaikawin⁶  Supachai Sakkhachornphop⁷  Methichit Wattanapanitch⁸  Koollawat Chupradit⁸ 

[1] Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand;Department of Anatomy, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand;Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand;Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA;Excellent Center for Drug Discovery, Mahidol University, Bangkok 10400, Thailand;Faculty of Medicine, King Mongkut’s Institute of Technology Ladkrabang, Bangkok 10520, Thailand;Research Institute for Health Sciences, Chiang Mai University, Chiang Mai 50200, Thailand;Siriraj Center for Regenerative Medicine, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand;

关键词: anti-HIV-1 scaffold;    zinc finger protein;    zinc finger domain;    hematopoietic stem/progenitor cell;    gene therapy;   

DOI  :  10.3390/ijms23042331

来源: DOAJ

【 摘 要 】

Human hematopoietic stem/progenitor cell (HSPC)-based gene therapy is a promising direction for curing HIV-1-infected individuals. The zinc finger protein (2LTRZFP) designed to target the 2-LTR-circle junction of HIV-1 cDNA was previously reported as an intracellular antiviral molecular scaffold that prevents HIV integration. Here, we elucidate the efficacy and safety of using 2LTRZFP in human CD34⁺ HSPCs. We transduced 2LTRZFP which has the mCherry tag (2LTRZFPmCherry) into human CD34⁺ HSPCs using a lentiviral vector. The 2LTRZFPmCherry-transduced HSPCs were subsequently differentiated into macrophages. The expression levels of pro-apoptotic proteins of the 2LTRZFPmCherry-transduced HSPCs showed no significant difference from those of the non-transduced control. Furthermore, the 2LTRZFPmCherry-transduced HSPCs were successfully differentiated into mature macrophages, which had normal phagocytic function. The cytokine secretion assay demonstrated that 2LTRZFPmCherry-transduced CD34⁺ derived macrophages promoted the polarization towards classically activated (M1) subtypes. More importantly, the 2LTRZFPmCherry transduced cells significantly exhibited resistance to HIV-1 integration in vitro. Our findings demonstrate that the 2LTRZFPmCherry-transduced macrophages were found to be functionally and phenotypically normal, with no adverse effects of the anti-HIV-1 scaffold. Our data suggest that the anti-HIV-1 integrase scaffold is a promising antiviral molecule that could be applied to human CD34⁺ HSPC-based gene therapy for AIDS patients.

【 授权许可】

Unknown