Service Project # 7: Lentivirus-like Particle Specific Delivery of Cas12 Ribonucleoprotein (RNP) to HIV Reservoir Cells in vivo for an HIV Cure 

Collaborating Investigator: Wenhui Hu
Affiliation: Temple University
Funding Status: NIH R01AI174301
Project Period: 12/01/22-02/28/27

Summary

The goal of SP #7 is intracellular delivery of mRNA and CRISPR/Cas genome editors for HIV treatment. This requires a delivery system for CD4+ T cell targeting and efficient intracellular delivery. NIMs developed in TR&D2 Aim 3 for gene delivery to T cells will be used to overcome this delivery challenge. Polymers will be sent to Dr. Hu to encapsulate mRNA and the CRISPR/Cas genome editors into nanoparticles, which will then be evaluated by Dr. Hu in his models. This academic research will be published and disseminated. Success may also lead to additional SPs requesting the validated TR&D2 materials for other CD4+ T cell targeting or gene editing uses. 

Approach

Aim. Evaluation of NIMs for CD4+ targeted gene delivery and gene editing.  This aim is focused on evaluation of the nanomaterials for gene delivery to CD4+ T cells. A CD4-targeted DARPin developed by Dr. Hu will be utilized to conjugate to the surface of leading NIMs for targeting.  GFP mRNA delivery will be evaluated first, and with positive results, Cas12a RNP/mRNA will be delivered subsequently. Initially experiments with several NIM formulations will be evaluated in vitro, with the best formulation then evaluated in vivo in mice. Ultimately, the technology is planned to be used to excise HIV provirus and CCR5 as a novel strategy for HIV treatment. This SP will validate the effectiveness of NIM-based materials for CD4+ T cell targeting and for gene editing applications. 

NIMs will be synthesized as described in TR&D2 Specific Aim 1, conjugated with the CD4-conjugated DARPin following the procedures of TR&D2 Specific Aim 3 and then shipped to Temple for evaluation.  The NIM use will be evaluated and optimized for mRNA and RNP/mRNA delivery to CD4+ T cells in vitro in multiwell plates and in vivo in mice following systemic injection.