Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the <i>CCR5</i> Locus
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 infor...
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MDPI AG
2023-01-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/1/202 |
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| author | Frederik Holm Rothemejer Nanna Pi Lauritsen Anna Karina Juhl Mariane Høgsbjerg Schleimann Saskia König Ole Schmeltz Søgaard Rasmus O. Bak Martin Tolstrup |
| author_facet | Frederik Holm Rothemejer Nanna Pi Lauritsen Anna Karina Juhl Mariane Høgsbjerg Schleimann Saskia König Ole Schmeltz Søgaard Rasmus O. Bak Martin Tolstrup |
| author_sort | Frederik Holm Rothemejer |
| collection | DOAJ |
| description | 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 <i>CCR5</i> 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. |
| first_indexed | 2024-03-09T11:01:51Z |
| format | Article |
| id | doaj.art-1753062e6ff145ec83a76b5f278a5b9e |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T11:01:51Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-1753062e6ff145ec83a76b5f278a5b9e2023-12-01T01:12:46ZengMDPI AGViruses1999-49152023-01-0115120210.3390/v15010202Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the <i>CCR5</i> LocusFrederik Holm Rothemejer0Nanna Pi Lauritsen1Anna Karina Juhl2Mariane Høgsbjerg Schleimann3Saskia König4Ole Schmeltz Søgaard5Rasmus O. Bak6Martin Tolstrup7Department of Clinical Medicine, Aarhus University, 8200 Aarhus, DenmarkDepartment of Clinical Medicine, Aarhus University, 8200 Aarhus, DenmarkDepartment of Clinical Medicine, Aarhus University, 8200 Aarhus, DenmarkDepartment of Clinical Medicine, Aarhus University, 8200 Aarhus, DenmarkDepartment of Biomedicine, Aarhus University, 8200 Aarhus, DenmarkDepartment of Clinical Medicine, Aarhus University, 8200 Aarhus, DenmarkDepartment of Biomedicine, Aarhus University, 8200 Aarhus, DenmarkDepartment of Clinical Medicine, Aarhus University, 8200 Aarhus, DenmarkAdoptive 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 <i>CCR5</i> 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.https://www.mdpi.com/1999-4915/15/1/202HIVCAR T cellsCRISPR/Cas9humanized mice |
| spellingShingle | Frederik Holm Rothemejer Nanna Pi Lauritsen Anna Karina Juhl Mariane Høgsbjerg Schleimann Saskia König Ole Schmeltz Søgaard Rasmus O. Bak Martin Tolstrup Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the <i>CCR5</i> Locus Viruses HIV CAR T cells CRISPR/Cas9 humanized mice |
| title | Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the <i>CCR5</i> Locus |
| title_full | Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the <i>CCR5</i> Locus |
| title_fullStr | Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the <i>CCR5</i> Locus |
| title_full_unstemmed | Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the <i>CCR5</i> Locus |
| title_short | Development of HIV-Resistant CAR T Cells by CRISPR/Cas-Mediated CAR Integration into the <i>CCR5</i> Locus |
| title_sort | development of hiv resistant car t cells by crispr cas mediated car integration into the i ccr5 i locus |
| topic | HIV CAR T cells CRISPR/Cas9 humanized mice |
| url | https://www.mdpi.com/1999-4915/15/1/202 |
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