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

doi:10.3390/v15010202

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 newspaper › Journal article › Research › peer-review

11 Citations (Scopus)

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 language	English
Article number	202
Journal	Viruses
Volume	15
Issue	1
Number of pages	13
ISSN	1999-4915
DOIs	https://doi.org/10.3390/v15010202
Publication status	Published - Jan 2023

Keywords

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

Access to Document

10.3390/v15010202Licence: CC BY

Cite this

@article{5fd8a15e8bbf4d619d7c457bfd30d489,

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

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. ",

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

author = "Rothemejer, {Frederik Holm} and Lauritsen, {Nanna Pi} and Juhl, {Anna Karina} and Schleimann, {Mariane H{\o}gsbjerg} and Saskia K{\"o}nig and S{\o}gaard, {Ole Schmeltz} and Rasmus Bak and Martin Tolstrup",

year = "2023",

month = jan,

doi = "10.3390/v15010202",

language = "English",

volume = "15",

journal = "Viruses",

issn = "1999-4915",

publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",

number = "1",

}

TY - JOUR

T1 - Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the CCR5 Locus

AU - Rothemejer, Frederik Holm

AU - Lauritsen, Nanna Pi

AU - Juhl, Anna Karina

AU - Schleimann, Mariane Høgsbjerg

AU - König, Saskia

AU - Søgaard, Ole Schmeltz

AU - Bak, Rasmus

AU - Tolstrup, Martin

PY - 2023/1

Y1 - 2023/1

N2 - 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.

AB - 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.

KW - Animals

KW - CRISPR-Cas Systems

KW - HIV Antibodies

KW - HIV Infections

KW - HIV-1

KW - Leukocytes, Mononuclear/metabolism

KW - Mice

KW - T-Lymphocytes

UR - http://www.scopus.com/inward/record.url?scp=85152284423&partnerID=8YFLogxK

U2 - 10.3390/v15010202

DO - 10.3390/v15010202

M3 - Journal article

C2 - 36680242

SN - 1999-4915

VL - 15

JO - Viruses

JF - Viruses

IS - 1

M1 - 202

ER -

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

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this