Pathways Toward a Functional HIV-1 Cure: Balancing Promise and Perils of CRISPR Therapy

Herskovitz, Jonathan; Hasan, Mahmudul; Patel, Milankumar; Kevadiya, Bhavesh D.; Gendelman, Howard E.

doi:10.1007/978-1-0716-1871-4_27

Cited by 3 publications

(3 citation statements)

References 81 publications

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“…Since the discovery of the CRISPR/Cas9 system it has been extensively studied regarding its application to inactivate HIV-1 29 . Identification of efficacious gRNA target sites that are conserved among divergent viral strains and multiplexing of gRNAs are key objectives for CRISPR/Cas therapy against HIV-1.…”

Section: Discussionmentioning

confidence: 99%

“…HIV-1 provirus excision or inactivation by genome editing technologies such as ZFNs, TALENs and the CRISPR/Cas system is one of the major HIV-1 eradication strategies. Besides targeting of the latent provirus, the CRISPR/Cas system has also been investigated as a therapeutic approach to immunize cells against HIV-1 infection and/or suppress viral replication 29 .…”

Section: Discussionmentioning

confidence: 99%

“…HIV-1 targeting with CRISPR/Cas is one of the major HIV-1 eradication strategies and has been investigated in numerous studies using Staphylococcus pyogenes Cas9, Staphylococcus aureus Cas9, Cas12a or Cas13a, and targeting different regulatory or structural genome region such as the LTR, gag , pol , tat and rev genes 9 . Furthermore, the CRISPR/Cas system can also be harnessed as a therapy during acute HIV-1 infection to inhibit and suppress virus replication while simultaneously targeting integrated HIV-1 proviruses 10,11 .…”

Section: Introductionmentioning

confidence: 99%

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Targeting HIV-1 with CRISPR/Cas9 delivered by retargeted adenoviruses effectively suppresses viral replication

Klinnert,

Freitag,

Plückthun

et al. 2023

Preprint

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Integrated, intact, latent HIV-1 viruses in infected cells are the main obstacle to curing HIV-1 infections. Targeted inactivation of HIV-1 proviruses with CRISPR/Cas9 is a promising strategy to eradicate HIV-1. In addition, CRISPR/Cas9 is able to target replicating HIV-1 and could be used as a therapy during productive infection.Here, we combine the CRISPR/Cas9 system with a novel adenovirus (Ad) targeted delivery technology to test it as a therapeutic approach to inhibit HIV-1. First, we selected six HIV-1-specific gRNAs targeting the HIV-1 LTRs and thegaggene and tested their efficacy in inhibiting HIV-1 virion production in an HEK 293T cell co-transfection screen. The gRNA-TAR showed the most robust and potent inhibition of HIV-1 by >99% alone or in combination with the gRNA-p24, which induced a ∼1 kb deletion between both gRNA target sites in HIV-1 DNA. Delivery of this dual gRNA-TAR/p24 CRISPR/Cas9 system with CD3-CD28-IL2-retargeted Ads was highly effective, transducing 62.3±23.3% of cells and suppressing HIV-1 replication by 88.0±4.5% in primary CD4+T cells from three independent donors.Our dual gRNA-TAR/p24-CRISPR/Cas9-Ad strategy represents a novel therapeutic approach to effectively inhibit HIV-1 in a highly HIV-1 and T cell-specific manner.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Targeting HIV-1 with CRISPR/Cas9 delivered by retargeted adenoviruses effectively suppresses viral replication

Klinnert,

Freitag,

Plückthun

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

Latent HIV-1 provirus in vitro suppression using combinatorial CRISPR/Cas9 strategy

et al. 2022

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Targeted shock-and-kill HIV-1 gene therapy approach combining CRISPR activation, suicide gene tBid and retargeted adenovirus delivery

et al. 2023

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Infections with the human immunodeficiency virus type 1 (HIV-1) are incurable due the long-lasting, latent viral reservoir. The shock-and-kill cure approach aims to activate latent proviruses in HIV-1 infected cells and subsequently kill these cells with strategies such as therapeutic vaccines or immune enhancement. Here, we combined the dCas9-VPR CRISPR activation (CRISPRa) system with gRNA-V, the truncated Bid (tBid)-based suicide gene strategy and CD3-retargeted adenovirus (Ad) delivery vectors, in an all-in-one targeted shock-and-kill gene therapy approach to achieve specific elimination of latently HIV-1 infected cells. Simultaneous transduction of latently HIV-1 infected J-Lat 10.6 cells with a CD3-retargeted Ad-CRISPRa-V and Ad-tBid led to a 57.7 ± 17.0% reduction of productively HIV-1 infected cells and 2.4-fold ± 0.25 increase in cell death. The effective activation of latent HIV-1 provirus by Ad-CRISPRa-V was similar to the activation control TNF-α. The strictly HIV-1 dependent and non-leaky killing by tBid could be demonstrated. Furthermore, the high transduction efficiencies of up to 70.8 ± 0.4% by the CD3-retargeting technology in HIV-1 latently infected cell lines was the basis of successful shock-and-kill. This novel targeted shock-and-kill all-in-one gene therapy approach has the potential to safely and effectively eliminate HIV-1 infected cells in a highly HIV-1 and T cell specific manner.

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Pathways Toward a Functional HIV-1 Cure: Balancing Promise and Perils of CRISPR Therapy

Cited by 3 publications

References 81 publications

Targeting HIV-1 with CRISPR/Cas9 delivered by retargeted adenoviruses effectively suppresses viral replication

Targeting HIV-1 with CRISPR/Cas9 delivered by retargeted adenoviruses effectively suppresses viral replication

Latent HIV-1 provirus in vitro suppression using combinatorial CRISPR/Cas9 strategy

Targeted shock-and-kill HIV-1 gene therapy approach combining CRISPR activation, suicide gene tBid and retargeted adenovirus delivery

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