CXCR4 Cell and Lymphoid Targeted CRISPR-Cas9 Lipid Nanoparticles Facilitate HIV-1 Proviral DNA Excision in CD4+ T cells

Author

Lubaba A. Zaman, University of Nebraska Medical CenterFollow

ORCID ID

0009-0001-4600-6411

Graduation Date

Spring 5-10-2025

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Programs

Interdisciplinary Graduate Program in Biomedical Sciences

First Advisor

Howard E Gendelman

Abstract

Advancements in antiretroviral therapy (ART) have enabled those people living with HIV-1 (PLWH) to live a life free from the many complications of viral infection. However, the daily use of medications for the person’s lifetime and the associated social stigma are impactful for cost and access. Moreover, type one HIV (HIV-1) is latent in multiple tissue and cell reservoirs resulting in viral rebound and the emergence of viral resistance. The latter is followed by ART interruption. However, to date, a ‘cure’ for HIV-1 remains elusive. With these needs in mind, we have developed targeted lipid nanoparticles (T-LNPs) as a means to improve delivery that can lead to viral elimination. Although viral and nanoparticle approaches were developed to deliver clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 formulations, a safe and effective means to preclude off-target toxicities is not available. Changing the chemical composition and surface functionality of LNPs can selectively deliver CRISPR/Cas9-guide RNA (gRNA) complex. We demonstrated that LNPs that target the CXCR4 receptor can selectively excise integrated viral DNA in laboratory cell models and primary CD4+ T cells, therefore, can be used for delivery to HIV-infected cells and tissues in animal models of persistent human infection. Moreover, we altered the tropism of LNPs to deliver to extrahepatic organs, specifically the spleen. Therefore, it can be tuned to effectively deliver to HIV-1 organ reservoirs as well. Finally, we have shown that that this two-pronged approach can excise viral DNA from infected humanized mice. In summary, our studies provided a proof-of- concept that we can deliver gene-editing therapies safely and effectively to difficult-to-reach latent HIV-1 reservoirs for viral DNA excision.

Comments

2025 Copyright, the authors

Recommended Citation

Zaman, Lubaba A., "CXCR4 Cell and Lymphoid Targeted CRISPR-Cas9 Lipid Nanoparticles Facilitate HIV-1 Proviral DNA Excision in CD4+ T cells" (2025). Theses & Dissertations. 921.
https://digitalcommons.unmc.edu/etd/921