# CD4 T cell-targeted nanoparticle in vivo delivery of CRISPR/Cas9 genome editors for HIV cure

> **NIH NIH R01** · TEMPLE UNIV OF THE COMMONWEALTH · 2021 · $753,945

## Abstract

SUMMARY
 HIV-1 infection requires a life-term antiretroviral treatment because its cessation leads to rapid viral
rebound from the HIV-1 latent cellular/tissue reservoir. Novel approaches to eradicate or permanently silence
HIV-1 proviruses are urgently needed to achieve a “sterile” cure of HIV infection, for which CRIPSR/Cas9
genome editing has opened a new avenue. In the past years, we and others have utilized Cas9-mediated
genome editing to excise HIV-1 provirus in vitro, ex vivo and in vivo. One of challenges before clinical
application is how to deliver effectively, specifically and safely the powerful genome editing machinery to HIV-1
latently infected cells. The objective of this proposal is to develop novel synthetic nanoparticle (NP) for the in
vivo delivery of Cas9/sgRNA ribonucleoprotein (RNP) specifically to CD4 T cells, the most important HIV-1
latent cellular reservoir. We have recently developed a novel synthetic PEG-Morpholine copolymer (PEG-pMor)
NP system for in vivo drug delivery in mouse model. Our preliminary data demonstrated the feasibility and
efficiency of this PEG-pMor NP to deliver Cas9/sgRNA plasmid or RNP in multiple organs/tissues resulting in
eradication of HIV-1 proviral DNA or host cellular genes in vivo. In this proposal, a novel CD4-specific designed
ankyrin repeat protein (DARPin) peptide will be displayed on the surface of the PEG-pMor NP to achieve
targeted delivery of Cas9/sgRNA RNP to human CD4 T cells in HIV-1-infected humanized mouse models. In
Aim I, we will develop and characterize CD4 T cell-targeting NP both in vitro and in vivo and determine the
efficiency of CD4-specific DARPin-mediated Cas9/duplex sgRNA RNP to excise CCR5 gene in human primary
T cells (in vitro) and humanized mouse model (in vivo). In Aim II, we will determine the efficiency of CD4 T cell-
targeting NP for in vivo delivery of Cas9/sgRNA RNP to excise HIV-1 proviral DNA. In Aim III, we will assess
the combinatory therapeutic potential of CD4 T cell-targeting NP in vivo delivery of Cas9/quadruplex sgRNA
RNP (LTR1/GagD+CCR5-A/B) in blocking or delaying latent HIV-1 viral rebound in humanized mouse model.
This high-reward proposal focuses on the screening of novel CD4 T cell-specific delivery of RNP genome
editors to excise HIV-1 provirus and disable HIV-1 entry coreceptor CCR5 gene. The positive outcome will
offer a novel tool to deliver Cas9/sgRNA RNP to CD4 T cells and/or other reservoir cells in vivo, and thus
provide new avenues for the development of therapeutics to cure HIV-1.

## Key facts

- **NIH application ID:** 10129897
- **Project number:** 5R01AI145034-03
- **Recipient organization:** TEMPLE UNIV OF THE COMMONWEALTH
- **Principal Investigator:** Wenhui Hu
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $753,945
- **Award type:** 5
- **Project period:** 2019-04-08 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10129897

## Citation

> US National Institutes of Health, RePORTER application 10129897, CD4 T cell-targeted nanoparticle in vivo delivery of CRISPR/Cas9 genome editors for HIV cure (5R01AI145034-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10129897. Licensed CC0.

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