Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the CCR5 Locus

Frederik Holm Rothemejer, Nanna Pi Lauritsen, Anna Karina Juhl, Mariane Høgsbjerg Schleimann, Saskia König, Ole Schmeltz Søgaard, Rasmus Bak, Martin Tolstrup*

*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

Adoptive immunotherapy using chimeric antigen receptor (CAR) T cells has been highly successful in treating B cell malignancies and holds great potential as a curative strategy for HIV infection. Recent advances in the use of anti-HIV broadly neutralizing antibodies (bNAbs) have provided vital information for optimal antigen targeting of CAR T cells. However, CD4+ CAR T cells are susceptible to HIV infection, limiting their therapeutic potential. In the current study, we engineered HIV-resistant CAR T cells using CRISPR/Cas9-mediated integration of a CAR cassette into the CCR5 locus. We used a single chain variable fragment (scFv) of the clinically potent bNAb 10-1074 as the antigen-targeting domain in our anti-HIV CAR T cells. Our anti-HIV CAR T cells showed specific lysis of HIV-infected cells in vitro. In a PBMC humanized mouse model of HIV infection, the anti-HIV CAR T cells expanded and transiently limited HIV infection. In conclusion, this study provides proof-of-concept for developing HIV-resistant CAR T cells using CRISPR/Cas9 targeted integration.

Original languageEnglish
Article number202
JournalViruses
Volume15
Issue1
Number of pages13
ISSN1999-4915
DOIs
Publication statusPublished - Jan 2023

Keywords

  • Mice
  • Animals
  • HIV Infections
  • CRISPR-Cas Systems
  • Leukocytes, Mononuclear/metabolism
  • HIV Antibodies
  • HIV-1
  • T-Lymphocytes

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