Development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune-checkpoint TIGIT
IntroductionT cell Ig and ITIM domain receptor (TIGIT) is a next-generation immune checkpoint predominantly expressed on activated T cells and NK cells, exhibiting an unfavorable prognostic association with various malignancies. Despite the emergence of multiple TIGIT-blocking agents entering clinic...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-09-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1268900/full |
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| author | Katty Zeven Timo W.M. De Groof Hannelore Ceuppens Robin Maximilian Awad Thomas Ertveldt Wout de Mey Fien Meeus Geert Raes Geert Raes Karine Breckpot Nick Devoogdt |
| author_facet | Katty Zeven Timo W.M. De Groof Hannelore Ceuppens Robin Maximilian Awad Thomas Ertveldt Wout de Mey Fien Meeus Geert Raes Geert Raes Karine Breckpot Nick Devoogdt |
| author_sort | Katty Zeven |
| collection | DOAJ |
| description | IntroductionT cell Ig and ITIM domain receptor (TIGIT) is a next-generation immune checkpoint predominantly expressed on activated T cells and NK cells, exhibiting an unfavorable prognostic association with various malignancies. Despite the emergence of multiple TIGIT-blocking agents entering clinical trials, only a fraction of patients responded positively to anti-TIGIT therapy. Consequently, an urgent demand arises for noninvasive techniques to quantify and monitor TIGIT expression, facilitating patient stratification and enhancing therapeutic outcomes. Small antigen binding moieties such as nanobodies, are promising candidates for such tracer development.MethodsWe generated a panel of anti-human or anti-mouse TIGIT nanobodies from immunized llamas. In addition, we designed a single-chain variable fragment derived from the clinically tested monoclonal antibody Vibostolimab targeting TIGIT, and assessed its performance alongside the nanobodies. In vitro characterization studies were performed, including binding ability and affinity to cell expressed or recombinant TIGIT. After Technetium-99m labeling, the nanobodies and the single-chain variable fragment were evaluated in vivo for their ability to detect TIGIT expression using SPECT/CT imaging, followed by ex vivo biodistribution analysis.ResultsNine nanobodies were selected for binding to recombinant and cell expressed TIGIT with low sub-nanomolar affinities and are thermostable. A six-fold higher uptake in TIGIT-overexpressing tumor was demonstrated one hour post- injection with Technetium-99m labeled nanobodies compared to an irrelevant control nanobody. Though the single-chain variable fragment exhibited superior binding to TIGIT-expressing peripheral blood mononuclear cells in vitro, its in vivo behavior yielded lower tumor-to-background ratios at one hour post- injection, indicating that nanobodies are better suited for in vivo imaging than the single-chain variable fragment. Despite the good affinity, high specificity and on-target uptake in mice in this setting, imaging of TIGIT expression on tumor- infiltrating lymphocytes within MC38 tumors remained elusive. This is likely due to the low expression levels of TIGIT in this model.DiscussionThe excellent affinity, high specificity and rapid on-target uptake in mice bearing TIGIT- overexpressing tumors showed the promising diagnostic potential of nanobodies to noninvasively image high TIGIT expression within the tumor. These findings hold promise for clinical translation to aid patient selection and improve therapy response. |
| first_indexed | 2024-03-11T23:11:08Z |
| format | Article |
| id | doaj.art-051a7b27d7524e158560ba763978542a |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-03-11T23:11:08Z |
| publishDate | 2023-09-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj.art-051a7b27d7524e158560ba763978542a2023-09-21T08:27:33ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-09-011410.3389/fimmu.2023.12689001268900Development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune-checkpoint TIGITKatty Zeven0Timo W.M. De Groof1Hannelore Ceuppens2Robin Maximilian Awad3Thomas Ertveldt4Wout de Mey5Fien Meeus6Geert Raes7Geert Raes8Karine Breckpot9Nick Devoogdt10Laboratory of Molecular Imaging and Therapy (MITH), Vrije Universiteit Brussel (VUB), Brussels, BelgiumLaboratory of Molecular Imaging and Therapy (MITH), Vrije Universiteit Brussel (VUB), Brussels, BelgiumLaboratory for Molecular and Cellular Therapy (LMCT), Department of Biomedical Sciences, Vrije Universiteit Brussel (VUB), Brussels, BelgiumLaboratory for Molecular and Cellular Therapy (LMCT), Department of Biomedical Sciences, Vrije Universiteit Brussel (VUB), Brussels, BelgiumLaboratory for Molecular and Cellular Therapy (LMCT), Department of Biomedical Sciences, Vrije Universiteit Brussel (VUB), Brussels, BelgiumLaboratory for Molecular and Cellular Therapy (LMCT), Department of Biomedical Sciences, Vrije Universiteit Brussel (VUB), Brussels, BelgiumLaboratory for Molecular and Cellular Therapy (LMCT), Department of Biomedical Sciences, Vrije Universiteit Brussel (VUB), Brussels, BelgiumLaboratory for Cellular and Molecular Immunology (CMIM), Vrije Universiteit Brussel (VUB), Brussels, BelgiumMyeloid Cell Immunology Lab, Vlaams Instituut voor Biotechnologie (VIB) Center for Inflammation Research, Brussels, BelgiumLaboratory for Molecular and Cellular Therapy (LMCT), Department of Biomedical Sciences, Vrije Universiteit Brussel (VUB), Brussels, BelgiumLaboratory of Molecular Imaging and Therapy (MITH), Vrije Universiteit Brussel (VUB), Brussels, BelgiumIntroductionT cell Ig and ITIM domain receptor (TIGIT) is a next-generation immune checkpoint predominantly expressed on activated T cells and NK cells, exhibiting an unfavorable prognostic association with various malignancies. Despite the emergence of multiple TIGIT-blocking agents entering clinical trials, only a fraction of patients responded positively to anti-TIGIT therapy. Consequently, an urgent demand arises for noninvasive techniques to quantify and monitor TIGIT expression, facilitating patient stratification and enhancing therapeutic outcomes. Small antigen binding moieties such as nanobodies, are promising candidates for such tracer development.MethodsWe generated a panel of anti-human or anti-mouse TIGIT nanobodies from immunized llamas. In addition, we designed a single-chain variable fragment derived from the clinically tested monoclonal antibody Vibostolimab targeting TIGIT, and assessed its performance alongside the nanobodies. In vitro characterization studies were performed, including binding ability and affinity to cell expressed or recombinant TIGIT. After Technetium-99m labeling, the nanobodies and the single-chain variable fragment were evaluated in vivo for their ability to detect TIGIT expression using SPECT/CT imaging, followed by ex vivo biodistribution analysis.ResultsNine nanobodies were selected for binding to recombinant and cell expressed TIGIT with low sub-nanomolar affinities and are thermostable. A six-fold higher uptake in TIGIT-overexpressing tumor was demonstrated one hour post- injection with Technetium-99m labeled nanobodies compared to an irrelevant control nanobody. Though the single-chain variable fragment exhibited superior binding to TIGIT-expressing peripheral blood mononuclear cells in vitro, its in vivo behavior yielded lower tumor-to-background ratios at one hour post- injection, indicating that nanobodies are better suited for in vivo imaging than the single-chain variable fragment. Despite the good affinity, high specificity and on-target uptake in mice in this setting, imaging of TIGIT expression on tumor- infiltrating lymphocytes within MC38 tumors remained elusive. This is likely due to the low expression levels of TIGIT in this model.DiscussionThe excellent affinity, high specificity and rapid on-target uptake in mice bearing TIGIT- overexpressing tumors showed the promising diagnostic potential of nanobodies to noninvasively image high TIGIT expression within the tumor. These findings hold promise for clinical translation to aid patient selection and improve therapy response.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1268900/fullTIGITimmune checkpoint (ICP)nuclear imagingnoninvasive diagnosistracer developmentnanobodies |
| spellingShingle | Katty Zeven Timo W.M. De Groof Hannelore Ceuppens Robin Maximilian Awad Thomas Ertveldt Wout de Mey Fien Meeus Geert Raes Geert Raes Karine Breckpot Nick Devoogdt Development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune-checkpoint TIGIT Frontiers in Immunology TIGIT immune checkpoint (ICP) nuclear imaging noninvasive diagnosis tracer development nanobodies |
| title | Development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune-checkpoint TIGIT |
| title_full | Development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune-checkpoint TIGIT |
| title_fullStr | Development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune-checkpoint TIGIT |
| title_full_unstemmed | Development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune-checkpoint TIGIT |
| title_short | Development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune-checkpoint TIGIT |
| title_sort | development and evaluation of nanobody tracers for noninvasive nuclear imaging of the immune checkpoint tigit |
| topic | TIGIT immune checkpoint (ICP) nuclear imaging noninvasive diagnosis tracer development nanobodies |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1268900/full |
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