Application of CRISPR/Cas Genomic Editing Tools for HIV Therapy: Toward Precise Modifications and Multilevel Protection

Abstract

Although highly active antiretroviral therapy (HAART) can robustly control human immunodeficiency virus (HIV) infection, the existence of latent HIV in a form of proviral DNA integrated into the host genome makes the virus insensitive to HAART. This requires patients to adhere to HAART for a lifetime, often leading to drug toxicity or viral resistance to therapy. Current genome-editing technologies offer different strategies to reduce the latent HIV reservoir in the body. In this review, we systematize the research on CRISPR/Cas-based anti-HIV therapeutic methods, discuss problems related to viral escape and gene editing, and try to focus on the technologies that effectively and precisely introduce genetic modifications and confer strong resistance to HIV infection. Particularly, knock-in (KI) approaches, such as mature B cells engineered to produce broadly neutralizing antibodies, T cells expressing fusion inhibitory peptides in the context of inactivated viral coreceptors, or provirus excision using base editors, look very promising. Current and future advancements in the precision of CRISPR/Cas editing and its delivery will help extend its applicability to clinical HIV therapy.

Increasing Gene Editing Efficiency via CRISPR/Cas9- or Cas12a-Mediated Knock-In in Primary Human T Cells

“…The second approach includes the introduction of new constructs into the genome to supply T cells with factors preventing viral entry or replication [37,38]. A recent review provides the details of these strategies [39].…”

CRISPR-Cas: ‘The Multipurpose Molecular Tool’ for Gene Therapy and Diagnosis

“…To date, three clinical trials have been set up using the ZFN method; one of them was carried out on T cells isolated from HIV-positive patients [ 7 ]. Chimeric ZFN proteins were engineered to modify the expression of CCR5 chemokine receptors, making them resistant to viral infection.…”

Novel perspectives on antisense transcription in HIV-1, HTLV-1, and HTLV-2

“…While highly active antiretroviral therapy (HAART) drugs have remained the mainstay of HIV-1 treatment, these drugs act by suppressing HIV-1 replication, and do not effectively eradicate HIV-1 proviral DNA from latently infected cells. Once HAART therapy is ceased, the latent infection is able to rebound and continue to debilitate patients (Eggleton and Nagalli, 2022;Maslennikova and Mazurov, 2022). Thus, the clear challenge in the race for an effective cure for HIV-1 infection lies in the removal of proviral DNA in latently infected cells, a similar challenge to that of HTLV-1 infection, and in fact all retroviral infections.…”

“…Thus, the clear challenge in the race for an effective cure for HIV-1 infection lies in the removal of proviral DNA in latently infected cells, a similar challenge to that of HTLV-1 infection, and in fact all retroviral infections. Multiple strategies exist for anti-HIV-1 applications of CRISPR, and they can be generally grouped into knock-out and knock-in strategies (Maslennikova and Mazurov, 2022). Knock-out of viral coreceptors CCR5 and CXCR4 and direct targeting of active and inactive proviral DNA exist as two strategies targeting the removal of critical stages of HIV-1 infection (Yu et al, 2018).…”

“…Off-target effects are also still a considerable drawback to all CRISPR therapeutic approaches. Much work is still needed to perfect these models for CRISPR therapeutics for the cure of HIV-1, and it is likely that HAART will remain first line for a considerable time (Maslennikova and Mazurov, 2022).…”