Knockout of the inhibitory receptor TIGIT enhances the antitumor response of ex vivo expanded NK cells and prevents fratricide with therapeutic Fc-active TIGIT antibodies
Background Inhibitory receptor T-cell Immunoreceptor with Ig and ITIM domains (TIGIT) expressed by Natural Killer (NK) and T cells regulates cancer immunity and has been touted as the next frontier in the development of cancer immunotherapeutics. Although early results of anti-TIGIT and its combinat...
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BMJ Publishing Group
2023-12-01
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| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/11/12/e007502.full |
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| author | Dean A Lee Jeremiah L Oyer Md Faqrul Hasan Tayler J Croom-Perez Liza D Robles-Carrillo Sanjana Kumar Brendan W Andersen Brian P Tullius Alicja J Copik Amanda R Campbell Thomas A Dieffenthaller Catherine A Cash Jonathan E Eloriaga Meisam Naeimi Kararoudi |
| author_facet | Dean A Lee Jeremiah L Oyer Md Faqrul Hasan Tayler J Croom-Perez Liza D Robles-Carrillo Sanjana Kumar Brendan W Andersen Brian P Tullius Alicja J Copik Amanda R Campbell Thomas A Dieffenthaller Catherine A Cash Jonathan E Eloriaga Meisam Naeimi Kararoudi |
| author_sort | Dean A Lee |
| collection | DOAJ |
| description | Background Inhibitory receptor T-cell Immunoreceptor with Ig and ITIM domains (TIGIT) expressed by Natural Killer (NK) and T cells regulates cancer immunity and has been touted as the next frontier in the development of cancer immunotherapeutics. Although early results of anti-TIGIT and its combinations with antiprogrammed death-ligand 1 were highly exciting, results from an interim analysis of phase III trials are disappointing. With mixed results, there is a need to understand the effects of therapeutic anti-TIGIT on the TIGIT+ immune cells to support its clinical use. Most of the TIGIT antibodies in development have an Fc-active domain, which binds to Fc receptors on effector cells. In mouse models, Fc-active anti-TIGIT induced superior immunity, while Fc receptor engagement was required for its efficacy. NK-cell depletion compromised the antitumor immunity of anti-TIGIT indicating the essential role of NK cells in the efficacy of anti-TIGIT. Since NK cells express TIGIT and Fc-receptor CD16, Fc-active anti-TIGIT may deplete NK cells via fratricide, which has not been studied.Methods CRISPR-Cas9-based TIGIT knockout (KO) was performed in expanded NK cells. Phenotypic and transcriptomic properties of TIGIT KO and wild-type (WT) NK cells were compared with flow cytometry, CyTOF, and RNA sequencing. The effect of TIGIT KO on NK-cell cytotoxicity was determined by calcein-AM release and live cell imaging-based cytotoxicity assays. The metabolic properties of TIGIT KO and WT NK cells were compared with a Seahorse analyzer. The effect of the Fc-component of anti-TIGIT on NK-cell fratricide was determined by co-culturing WT and TIGIT KO NK cells with Fc-active and Fc-inactive anti-TIGIT.Results TIGIT KO increased the cytotoxicity of NK cells against multiple cancer cell lines including spheroids. TIGIT KO NK cells upregulated mTOR complex 1 (mTORC1) signaling and had better metabolic fitness with an increased basal glycolytic rate when co-cultured with cancer cells compared with WT NK cells. Importantly, TIGIT KO prevented NK-cell fratricide when combined with Fc-active anti-TIGIT.Conclusions TIGIT KO in ex vivo expanded NK cells increased their cytotoxicity and metabolic fitness and prevented NK-cell fratricide when combined with Fc-active anti-TIGIT antibodies. These fratricide-resistant TIGIT KO NK cells have therapeutic potential alone or in combination with Fc-active anti-TIGIT antibodies to enhance their efficacy. |
| format | Article |
| id | doaj-art-4c6550d3cfe540f09ffbff19f9a789fa |
| institution | Kabale University |
| issn | 2051-1426 |
| language | English |
| publishDate | 2023-12-01 |
| publisher | BMJ Publishing Group |
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| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj-art-4c6550d3cfe540f09ffbff19f9a789fa2025-02-03T05:05:17ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262023-12-01111210.1136/jitc-2023-007502Knockout of the inhibitory receptor TIGIT enhances the antitumor response of ex vivo expanded NK cells and prevents fratricide with therapeutic Fc-active TIGIT antibodiesDean A Lee0Jeremiah L Oyer1Md Faqrul Hasan2Tayler J Croom-Perez3Liza D Robles-Carrillo4Sanjana Kumar5Brendan W Andersen6Brian P Tullius7Alicja J Copik8Amanda R Campbell9Thomas A Dieffenthaller10Catherine A Cash11Jonathan E Eloriaga12Meisam Naeimi Kararoudi13Abigail Wexner Research Institute at Nationwide Children`s Hospital, Columbus, Ohio, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USAPediatric Cellular Therapies, AdventHealth for Children, Orlando, Florida, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USAAbigail Wexner Research Institute at Nationwide Children`s Hospital, Columbus, Ohio, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USAAbigail Wexner Research Institute at Nationwide Children`s Hospital, Columbus, Ohio, USABurnett School of Biomedical Sciences, University of Central Florida, Orlando, Florida, USACenter for Childhood Cancer, Abigail Wexner Research Institute, Nationwide Children’s Hospital, Columbus, Ohio, USABackground Inhibitory receptor T-cell Immunoreceptor with Ig and ITIM domains (TIGIT) expressed by Natural Killer (NK) and T cells regulates cancer immunity and has been touted as the next frontier in the development of cancer immunotherapeutics. Although early results of anti-TIGIT and its combinations with antiprogrammed death-ligand 1 were highly exciting, results from an interim analysis of phase III trials are disappointing. With mixed results, there is a need to understand the effects of therapeutic anti-TIGIT on the TIGIT+ immune cells to support its clinical use. Most of the TIGIT antibodies in development have an Fc-active domain, which binds to Fc receptors on effector cells. In mouse models, Fc-active anti-TIGIT induced superior immunity, while Fc receptor engagement was required for its efficacy. NK-cell depletion compromised the antitumor immunity of anti-TIGIT indicating the essential role of NK cells in the efficacy of anti-TIGIT. Since NK cells express TIGIT and Fc-receptor CD16, Fc-active anti-TIGIT may deplete NK cells via fratricide, which has not been studied.Methods CRISPR-Cas9-based TIGIT knockout (KO) was performed in expanded NK cells. Phenotypic and transcriptomic properties of TIGIT KO and wild-type (WT) NK cells were compared with flow cytometry, CyTOF, and RNA sequencing. The effect of TIGIT KO on NK-cell cytotoxicity was determined by calcein-AM release and live cell imaging-based cytotoxicity assays. The metabolic properties of TIGIT KO and WT NK cells were compared with a Seahorse analyzer. The effect of the Fc-component of anti-TIGIT on NK-cell fratricide was determined by co-culturing WT and TIGIT KO NK cells with Fc-active and Fc-inactive anti-TIGIT.Results TIGIT KO increased the cytotoxicity of NK cells against multiple cancer cell lines including spheroids. TIGIT KO NK cells upregulated mTOR complex 1 (mTORC1) signaling and had better metabolic fitness with an increased basal glycolytic rate when co-cultured with cancer cells compared with WT NK cells. Importantly, TIGIT KO prevented NK-cell fratricide when combined with Fc-active anti-TIGIT.Conclusions TIGIT KO in ex vivo expanded NK cells increased their cytotoxicity and metabolic fitness and prevented NK-cell fratricide when combined with Fc-active anti-TIGIT antibodies. These fratricide-resistant TIGIT KO NK cells have therapeutic potential alone or in combination with Fc-active anti-TIGIT antibodies to enhance their efficacy.https://jitc.bmj.com/content/11/12/e007502.full |
| spellingShingle | Dean A Lee Jeremiah L Oyer Md Faqrul Hasan Tayler J Croom-Perez Liza D Robles-Carrillo Sanjana Kumar Brendan W Andersen Brian P Tullius Alicja J Copik Amanda R Campbell Thomas A Dieffenthaller Catherine A Cash Jonathan E Eloriaga Meisam Naeimi Kararoudi Knockout of the inhibitory receptor TIGIT enhances the antitumor response of ex vivo expanded NK cells and prevents fratricide with therapeutic Fc-active TIGIT antibodies Journal for ImmunoTherapy of Cancer |
| title | Knockout of the inhibitory receptor TIGIT enhances the antitumor response of ex vivo expanded NK cells and prevents fratricide with therapeutic Fc-active TIGIT antibodies |
| title_full | Knockout of the inhibitory receptor TIGIT enhances the antitumor response of ex vivo expanded NK cells and prevents fratricide with therapeutic Fc-active TIGIT antibodies |
| title_fullStr | Knockout of the inhibitory receptor TIGIT enhances the antitumor response of ex vivo expanded NK cells and prevents fratricide with therapeutic Fc-active TIGIT antibodies |
| title_full_unstemmed | Knockout of the inhibitory receptor TIGIT enhances the antitumor response of ex vivo expanded NK cells and prevents fratricide with therapeutic Fc-active TIGIT antibodies |
| title_short | Knockout of the inhibitory receptor TIGIT enhances the antitumor response of ex vivo expanded NK cells and prevents fratricide with therapeutic Fc-active TIGIT antibodies |
| title_sort | knockout of the inhibitory receptor tigit enhances the antitumor response of ex vivo expanded nk cells and prevents fratricide with therapeutic fc active tigit antibodies |
| url | https://jitc.bmj.com/content/11/12/e007502.full |
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