Engineered Zinc Finger Protein Targeting 2LTR Inhibits HIV Integration in Hematopoietic Stem and Progenitor...

Area of Science:

• Gene Therapy
• Virology
• Immunology

Background:

• Hematopoietic stem/progenitor cell (HSPC) gene therapy offers a potential cure for HIV-1.
• Zinc finger protein (2LTRZFP) targets HIV-1 cDNA's 2-LTR circle junction, inhibiting viral integration.

Purpose of the Study:

• To evaluate the efficacy and safety of 2LTRZFP in human CD34+ HSPCs for HIV-1 gene therapy.
• To assess the impact of 2LTRZFP transduction on HSPC differentiation and macrophage function.

Main Methods:

• Lentiviral vector-mediated transduction of 2LTRZFP with an mCherry tag (2LTRZFPmCherry) into human CD34+ HSPCs.
• Differentiation of transduced HSPCs into macrophages.
• Assessment of apoptosis, phagocytic function, cytokine secretion, and HIV-1 integration resistance in vitro.

Main Results:

• 2LTRZFPmCherry transduction did not significantly alter pro-apoptotic protein levels in HSPCs.
• Transduced HSPCs differentiated into mature macrophages with normal phagocytic capacity and M1 polarization.
• 2LTRZFPmCherry-transduced cells demonstrated significant resistance to HIV-1 integration in vitro.

Conclusions:

• 2LTRZFPmCherry-transduced macrophages are phenotypically and functionally normal, with no observed adverse effects.
• The anti-HIV-1 integrase scaffold (2LTRZFP) is a safe and effective antiviral molecule for HSPC-based gene therapy in HIV-1 patients.