Targeted Nanocarrier-Mediated Editing Efficiency and Tissue-Specific Clearance of HIV-1 Proviral DNA in Humanized Mouse Models

Efficient and tissue-specific elimination of HIV-1 proviral DNA remains a central challenge for curative strategies, particularly within lymphoid and sanctuary compartments. This study evaluated the delivery performance and editing outcomes of receptor-targeted nanocarriers designed to enhance proviral DNA disruption across heterogeneous cellular reservoirs. Humanized NOD/SCID/IL2Rγ-null mice (n = 54) chronically infected with HIV-1 and maintained under suppressive antiretroviral therapy were treated with engineered polymer–lipid hybrid nanoparticles encapsulating CRISPR-associated ribonucleoprotein complexes. Nanocarriers were functionalized to preferentially target CD4⁺ T cells and CXCR4-expressing myeloid populations. Biodistribution and cellular uptake were quantified using fluorescence-assisted tissue mapping and flow cytometry, while proviral DNA levels were measured by droplet digital PCR in lymph nodes, spleen, bone marrow, and brain at baseline and 8 weeks post-treatment. Targeted nanocarriers achieved a 2.1-fold increase in uptake within CXCR4⁺ cells compared with non-targeted controls (p < 0.01) and resulted in a 38–52% reduction of proviral DNA in lymphoid tissues. In contrast, non-lymphoid compartments exhibited limited editing efficiency (<15% reduction). Longitudinal modeling indicated that reservoir depletion was primarily driven by editing efficiency rather than changes in cell turnover. No significant off-target editing or systemic toxicity was detected. These results demonstrate that cell- and receptor-specific nanocarrier design substantially improves proviral DNA disruption in dominant HIV-1 reservoirs and provide quantitative benchmarks for evaluating targeted genome-editing approaches.