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  1. Article ; Online: Epigenetic reprogramming of CAR T cells for in vivo functional persistence against solid tumors.

    Saitakis, Michael

    Genes and immunity

    2024  

    Abstract: Limited CAR T-cell expansion and persistence hinder therapeutic responses in solid cancer patients. To enhance the functional persistence of engineered T-cell therapies, we performed genetic disruption in human CAR T cells of SUV39H1, a histone 3 lysine ... ...

    Abstract Limited CAR T-cell expansion and persistence hinder therapeutic responses in solid cancer patients. To enhance the functional persistence of engineered T-cell therapies, we performed genetic disruption in human CAR T cells of SUV39H1, a histone 3 lysine 9 methyltransferase that promotes heterochromatin formation. This resulted in phenotypic CAR-T reprogramming that elicited optimal and sustained antitumor functionality. Single-cell transcriptomic (scRNA-seq) and chromatin accessibility (scATAC-seq) analyses of tumor-infiltrating CAR T cells showed early reprogramming into self-renewing, stem-like populations with decreased expression of dysfunction genes in all subpopulations. Moreover, we provided evidence that SUV39H1 inactivation elicits potent and durable functional persistence upon multiple tumor rechallenges. This opens a safe path to enhancing adoptive cell therapies for solid tumors.
    Language English
    Publishing date 2024-02-22
    Publishing country England
    Document type Journal Article
    ZDB-ID 2060566-3
    ISSN 1476-5470 ; 1466-4879
    ISSN (online) 1476-5470
    ISSN 1466-4879
    DOI 10.1038/s41435-024-00262-x
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    Zs.A 5592: Show issues Location:
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  2. Article: Biophysical Aspects of T Lymphocyte Activation at the Immune Synapse.

    Hivroz, Claire / Saitakis, Michael

    Frontiers in immunology

    2016  Volume 7, Page(s) 46

    Abstract: T lymphocyte activation is a pivotal step of the adaptive immune response. It requires the recognition by T-cell receptors (TCR) of peptides presented in the context of major histocompatibility complex molecules (pMHC) present at the surface of antigen- ... ...

    Abstract T lymphocyte activation is a pivotal step of the adaptive immune response. It requires the recognition by T-cell receptors (TCR) of peptides presented in the context of major histocompatibility complex molecules (pMHC) present at the surface of antigen-presenting cells (APCs). T lymphocyte activation also involves engagement of costimulatory receptors and adhesion molecules recognizing ligands on the APC. Integration of these different signals requires the formation of a specialized dynamic structure: the immune synapse. While the biochemical and molecular aspects of this cell-cell communication have been extensively studied, its mechanical features have only recently been addressed. Yet, the immune synapse is also the place of exchange of mechanical signals. Receptors engaged on the T lymphocyte surface are submitted to many tensile and traction forces. These forces are generated by various phenomena: membrane undulation/protrusion/retraction, cell mobility or spreading, and dynamic remodeling of the actomyosin cytoskeleton inside the T lymphocyte. Moreover, the TCR can both induce force development, following triggering, and sense and convert forces into biochemical signals, as a bona fide mechanotransducer. Other costimulatory molecules, such as LFA-1, engaged during immune synapse formation, also display these features. Moreover, T lymphocytes themselves are mechanosensitive, since substrate stiffness can modulate their response. In this review, we will summarize recent studies from a biophysical perspective to explain how mechanical cues can affect T lymphocyte activation. We will particularly discuss how forces are generated during immune synapse formation; how these forces affect various aspects of T lymphocyte biology; and what are the key features of T lymphocyte response to stiffness.
    Language English
    Publishing date 2016
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2606827-8
    ISSN 1664-3224
    ISSN 1664-3224
    DOI 10.3389/fimmu.2016.00046
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  3. Article ; Online: SUV39H1 Ablation Enhances Long-term CAR T Function in Solid Tumors.

    López-Cobo, Sheila / Fuentealba, Jaime R / Gueguen, Paul / Bonté, Pierre-Emmanuel / Tsalkitzi, Kyriaki / Chacón, Irena / Glauzy, Salomé / Bohineust, Armelle / Biquand, Ariane / Silva, Lisseth / Gouveia, Zelia / Goudot, Christel / Perez, Franck / Saitakis, Michael / Amigorena, Sebastian

    Cancer discovery

    2023  Volume 14, Issue 1, Page(s) 120–141

    Abstract: Failure of adoptive T-cell therapies in patients with cancer is linked to limited T-cell expansion and persistence, even in memory-prone 41BB-(BBz)-based chimeric antigen receptor (CAR) T cells. We show here that BBz-CAR T-cell stem/memory ... ...

    Abstract Failure of adoptive T-cell therapies in patients with cancer is linked to limited T-cell expansion and persistence, even in memory-prone 41BB-(BBz)-based chimeric antigen receptor (CAR) T cells. We show here that BBz-CAR T-cell stem/memory differentiation and persistence can be enhanced through epigenetic manipulation of the histone 3 lysine 9 trimethylation (H3K9me3) pathway. Inactivation of the H3K9 trimethyltransferase SUV39H1 enhances BBz-CAR T cell long-term persistence, protecting mice against tumor relapses and rechallenges in lung and disseminated solid tumor models up to several months after CAR T-cell infusion. Single-cell transcriptomic (single-cell RNA sequencing) and chromatin opening (single-cell assay for transposase accessible chromatin) analyses of tumor-infiltrating CAR T cells show early reprogramming into self-renewing, stemlike populations with decreased expression of dysfunction genes in all T-cell subpopulations. Therefore, epigenetic manipulation of H3K9 methylation by SUV39H1 optimizes the long-term functional persistence of BBz-CAR T cells, limiting relapses, and providing protection against tumor rechallenges.
    Significance: Limited CAR T-cell expansion and persistence hinders therapeutic responses in solid cancer patients. We show that targeting SUV39H1 histone methyltransferase enhances 41BB-based CAR T-cell long-term protection against tumor relapses and rechallenges by increasing stemness/memory differentiation. This opens a safe path to enhancing adoptive cell therapies for solid tumors. See related article by Jain et al., p. 142. This article is featured in Selected Articles from This Issue, p. 5.
    MeSH term(s) Animals ; Humans ; Mice ; Chromatin ; Immunotherapy, Adoptive ; Methyltransferases/genetics ; Methyltransferases/metabolism ; Neoplasms/genetics ; Neoplasms/therapy ; Receptors, Chimeric Antigen ; Recurrence ; Repressor Proteins/genetics ; Repressor Proteins/metabolism
    Chemical Substances Chromatin ; Methyltransferases (EC 2.1.1.-) ; Receptors, Chimeric Antigen ; Repressor Proteins ; SUV39H1 protein, human (EC 2.1.1.) ; Suv39h1 protein, mouse (EC 2.1.1.)
    Language English
    Publishing date 2023-11-07
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2625242-9
    ISSN 2159-8290 ; 2159-8274
    ISSN (online) 2159-8290
    ISSN 2159-8274
    DOI 10.1158/2159-8290.CD-22-1350
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 6916: Show issues Location:
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  4. Article ; Online: Acoustic sensors as a biophysical tool for probing cell attachment and cell/surface interactions.

    Saitakis, Michael / Gizeli, Electra

    Cellular and molecular life sciences : CMLS

    2011  Volume 69, Issue 3, Page(s) 357–371

    Abstract: Acoustic biosensors offer the possibility to analyse cell attachment and spreading. This is due to the offered speed of detection, the real-time non-invasive approach and their high sensitivity not only to mass coupling, but also to viscoelastic changes ... ...

    Abstract Acoustic biosensors offer the possibility to analyse cell attachment and spreading. This is due to the offered speed of detection, the real-time non-invasive approach and their high sensitivity not only to mass coupling, but also to viscoelastic changes occurring close to the sensor surface. Quartz crystal microbalance (QCM) and surface acoustic wave (Love-wave) systems have been used to monitor the adhesion of animal cells to various surfaces and record the behaviour of cell layers under various conditions. The sensors detect cells mostly via their sensitivity in viscoelasticity and mechanical properties. Particularly, the QCM sensor detects cytoskeletal rearrangements caused by specific drugs affecting either actin microfilaments or microtubules. The Love-wave sensor directly measures cell/substrate bonds via acoustic damping and provides 2D kinetic and affinity parameters. Other studies have applied the QCM sensor as a diagnostic tool for leukaemia and, potentially, for chemotherapeutic agents. Acoustic sensors have also been used in the evaluation of the cytocompatibility of artificial surfaces and, in general, they have the potential to become powerful tools for even more diverse cellular analysis.
    MeSH term(s) Acoustics ; Animals ; Biosensing Techniques/instrumentation ; Cell Adhesion ; Cytoskeleton ; Humans ; Polymers/chemistry ; Quartz Crystal Microbalance Techniques ; Surface Properties
    Chemical Substances Polymers
    Language English
    Publishing date 2011-10-15
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 1358415-7
    ISSN 1420-9071 ; 1420-682X
    ISSN (online) 1420-9071
    ISSN 1420-682X
    DOI 10.1007/s00018-011-0854-8
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  5. Article ; Online: Quantification of the effect of glycocalyx condition on membrane receptor interactions using an acoustic wave sensor.

    Saitakis, Michael / Gizeli, Electra

    European biophysics journal : EBJ

    2010  Volume 40, Issue 2, Page(s) 209–215

    Abstract: The effect of the cell glycocalyx on the binding of a membrane receptor, class I major histocompatibility complex (MHC) human leukocyte antigen (HLA)-A2, to an immobilized anti-HLA antibody was investigated using an acoustic sensor based on a Love wave ... ...

    Abstract The effect of the cell glycocalyx on the binding of a membrane receptor, class I major histocompatibility complex (MHC) human leukocyte antigen (HLA)-A2, to an immobilized anti-HLA antibody was investigated using an acoustic sensor based on a Love wave geometry. The enzyme neuraminidase was used to remove sialic acid residues from the cell glycocalyx. Real-time measurements of the amplitude of the acoustic wave showed that treatment with neuraminidase facilitates HLA/anti-HLA-mediated cell attachment via a 3.6-fold increase of the two-dimensional (2D) binding constant of the interaction. This could be attributed to better approach of binding partners due to favorable condition of the desialylated glycocalyx. The results underline the importance of microtopological factors in membrane receptor binding and reveal the potential of the Love wave sensor and 2D binding parameters for studying cell-substrate binding events.
    MeSH term(s) Acoustics/instrumentation ; Binding Sites ; Biosensing Techniques/instrumentation ; Biosensing Techniques/methods ; Cell Membrane/metabolism ; Glycocalyx/metabolism ; HLA-A2 Antigen/chemistry ; HLA-A2 Antigen/metabolism ; Humans ; Kinetics ; Receptors, Cell Surface/chemistry ; Receptors, Cell Surface/metabolism
    Chemical Substances HLA-A2 Antigen ; Receptors, Cell Surface
    Language English
    Publishing date 2010-10-17
    Publishing country Germany
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 283671-3
    ISSN 1432-1017 ; 0175-7571
    ISSN (online) 1432-1017
    ISSN 0175-7571
    DOI 10.1007/s00249-010-0632-9
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  6. Article: Quantification of the effect of glycocalyx condition on membrane receptor interactions using an acoustic wave sensor

    Saitakis, Michael / Gizeli, Electra

    European biophysics journal with biophysics letters. 2011 Feb., v. 40, no. 2

    2011  

    Abstract: The effect of the cell glycocalyx on the binding of a membrane receptor, class I major histocompatibility complex (MHC) human leukocyte antigen (HLA)-A2, to an immobilized anti-HLA antibody was investigated using an acoustic sensor based on a Love wave ... ...

    Abstract The effect of the cell glycocalyx on the binding of a membrane receptor, class I major histocompatibility complex (MHC) human leukocyte antigen (HLA)-A2, to an immobilized anti-HLA antibody was investigated using an acoustic sensor based on a Love wave geometry. The enzyme neuraminidase was used to remove sialic acid residues from the cell glycocalyx. Real-time measurements of the amplitude of the acoustic wave showed that treatment with neuraminidase facilitates HLA/anti-HLA-mediated cell attachment via a 3.6-fold increase of the two-dimensional (2D) binding constant of the interaction. This could be attributed to better approach of binding partners due to favorable condition of the desialylated glycocalyx. The results underline the importance of microtopological factors in membrane receptor binding and reveal the potential of the Love wave sensor and 2D binding parameters for studying cell-substrate binding events.
    Language English
    Dates of publication 2011-02
    Size p. 209-215.
    Publisher Springer-Verlag
    Publishing place Berlin/Heidelberg
    Document type Article
    ZDB-ID 283671-3
    ISSN 1432-1017 ; 0175-7571
    ISSN (online) 1432-1017
    ISSN 0175-7571
    DOI 10.1007/s00249-010-0632-9
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  7. Article ; Online: In vivo genome-wide CRISPR screens identify SOCS1 as intrinsic checkpoint of CD4

    Sutra Del Galy, Aurélien / Menegatti, Silvia / Fuentealba, Jaime / Lucibello, Francesca / Perrin, Laetitia / Helft, Julie / Darbois, Aurélie / Saitakis, Michael / Tosello, Jimena / Rookhuizen, Derek / Deloger, Marc / Gestraud, Pierre / Socié, Gérard / Amigorena, Sebastian / Lantz, Olivier / Menger, Laurie

    Science immunology

    2021  Volume 6, Issue 66, Page(s) eabe8219

    Abstract: ... Although ... ...

    Abstract Although CD8
    MeSH term(s) Animals ; CD4-Positive T-Lymphocytes/immunology ; Clustered Regularly Interspaced Short Palindromic Repeats/immunology ; Female ; Male ; Mice ; Mice, Knockout ; Mice, Transgenic ; Suppressor of Cytokine Signaling 1 Protein/immunology ; Th1 Cells/immunology
    Chemical Substances Socs1 protein, mouse ; Suppressor of Cytokine Signaling 1 Protein
    Language English
    Publishing date 2021-12-03
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 2470-9468
    ISSN (online) 2470-9468
    DOI 10.1126/sciimmunol.abe8219
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  8. Article ; Online: Different TCR-induced T lymphocyte responses are potentiated by stiffness with variable sensitivity.

    Saitakis, Michael / Dogniaux, Stéphanie / Goudot, Christel / Bufi, Nathalie / Asnacios, Sophie / Maurin, Mathieu / Randriamampita, Clotilde / Asnacios, Atef / Hivroz, Claire

    eLife

    2017  Volume 6

    Abstract: T cells are mechanosensitive but the effect of stiffness on their functions is still debated. We characterize herein how human primary ... ...

    Abstract T cells are mechanosensitive but the effect of stiffness on their functions is still debated. We characterize herein how human primary CD4
    Language English
    Publishing date 2017--08
    Publishing country England
    Document type Journal Article
    ZDB-ID 2687154-3
    ISSN 2050-084X ; 2050-084X
    ISSN (online) 2050-084X
    ISSN 2050-084X
    DOI 10.7554/eLife.23190
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  9. Article ; Online: Qualitative differences in T-cell activation by dendritic cell-derived extracellular vesicle subtypes.

    Tkach, Mercedes / Kowal, Joanna / Zucchetti, Andres E / Enserink, Lotte / Jouve, Mabel / Lankar, Danielle / Saitakis, Michael / Martin-Jaular, Lorena / Théry, Clotilde

    The EMBO journal

    2017  Volume 36, Issue 20, Page(s) 3012–3028

    Abstract: Exosomes, nano-sized secreted extracellular vesicles (EVs), are actively studied for their diagnostic and therapeutic potential. In particular, exosomes secreted by dendritic cells (DCs) have been shown to carry MHC-peptide complexes allowing efficient ... ...

    Abstract Exosomes, nano-sized secreted extracellular vesicles (EVs), are actively studied for their diagnostic and therapeutic potential. In particular, exosomes secreted by dendritic cells (DCs) have been shown to carry MHC-peptide complexes allowing efficient activation of T lymphocytes, thus displaying potential as promoters of adaptive immune responses. DCs also secrete other types of EVs of different size, subcellular origin and protein composition, whose immune capacities have not been yet compared to those of exosomes. Here, we show that large EVs (lEVs) released by human DCs are as efficient as small EVs (sEVs), including exosomes, to induce CD4
    MeSH term(s) CD4-Positive T-Lymphocytes/drug effects ; CD4-Positive T-Lymphocytes/immunology ; Dendritic Cells/immunology ; Dendritic Cells/metabolism ; Extracellular Vesicles/metabolism ; Humans ; Lymphocyte Activation
    Language English
    Publishing date 2017-09-18
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 586044-1
    ISSN 1460-2075 ; 0261-4189
    ISSN (online) 1460-2075
    ISSN 0261-4189
    DOI 10.15252/embj.201696003
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  10. Article ; Online: Probing the interaction of a membrane receptor with a surface-attached ligand using whole cells on acoustic biosensors.

    Saitakis, Michael / Tsortos, Achilleas / Gizeli, Electra

    Biosensors & bioelectronics

    2010  Volume 25, Issue 7, Page(s) 1688–1693

    Abstract: Two different types of acoustic sensors, a surface acoustic wave device supporting a Love-wave (Love-SAW) and a quartz crystal microbalance system with dissipation (QCM-D), were used to demonstrate the potential of acoustic devices to probe the binding ... ...

    Abstract Two different types of acoustic sensors, a surface acoustic wave device supporting a Love-wave (Love-SAW) and a quartz crystal microbalance system with dissipation (QCM-D), were used to demonstrate the potential of acoustic devices to probe the binding of a cell membrane receptor to an immobilized ligand. The class I Major Histocompatibility Complex molecule HLA-A2 on the surface of whole cells and anti-HLA monoclonal antibodies immobilized on the sensor were used as an interaction pair. Acoustic measurements consisted of recording the energy and velocity or frequency of the acoustic wave. Results showed that both devices could detect the number of cells in solution as well as the cells bound to the surface. In addition, the Love-wave sensor, which can sense binding events within the relatively short distance of approximately 50 nm from the device surface, was sensitive to the number of bonds formed between the cell membrane and the device surface while the QCM-D, which can sense deeper within the liquid, was found to respond well to stimuli that affected the cell membrane rigidity (cytochalasin D treatment). The above results suggest that acoustic biosensors can be a powerful tool in the study of cell/substrate interactions and acoustic devices of different type can be used in a complementary way.
    MeSH term(s) Acoustics/instrumentation ; Biological Assay/instrumentation ; Biosensing Techniques/instrumentation ; Biosensing Techniques/methods ; Cell Membrane/metabolism ; Equipment Design ; Equipment Failure Analysis ; Ligands ; Molecular Probe Techniques/instrumentation ; Protein Binding ; Protein Interaction Mapping/instrumentation ; Receptors, Cell Surface/metabolism ; Reproducibility of Results ; Sensitivity and Specificity
    Chemical Substances Ligands ; Receptors, Cell Surface
    Language English
    Publishing date 2010-03-15
    Publishing country England
    Document type Evaluation Studies ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1011023-9
    ISSN 1873-4235 ; 0956-5663
    ISSN (online) 1873-4235
    ISSN 0956-5663
    DOI 10.1016/j.bios.2009.12.008
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