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Rapid Screening of CAR T Cell Functional Improvement Strategies by Highly Multiplexed Single-Cell Secretomics

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Cancer Immunotherapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2748))

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Abstract

The functional fitness of CAR T cells plays a crucial role in determining their clinical efficacy. Several strategies are being explored to increase cellular fitness, but screening these approaches in vivo is expensive and time-consuming, limiting the number of strategies that can be tested at one time. The presence of polyfunctional CAR T cells has emerged as a critical parameter correlating with clinical responses. However, even sophisticated multiplexed secretomic assays often fail to detect differences in cytokine release due to the functional heterogeneity of CAR T cell products. Here, we describe a highly multiplexed single-cell secretomic assay based on the IsoLight platform to rapidly evaluate the impact of new pharmacologic or gene-engineering approaches aiming at improving CAR T cell function. As a key study, we focus on CD19-specific CAR CD8+ T cells modulated by miR-155 overexpression, but the protocol can be applied to characterize other functional immune cell modulation strategies.

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Conflicting Interests

L.G. has consulting agreements with Lyell Immunopharma, Instil Bio, and Advaxis. L.G. is on the scientific advisory board of Poseida Therapeutics and Kiromic and a stockholder of Poseida Therapeutics. E.H. and J.Z. are employed by and have equity ownership in IsoPlexis.

Author information

Authors and Affiliations

  1. Division of Functional Immune Cell Modulation, Leibniz Institute for Immunotherapy (LIT), Regensburg, Germany

    Dragana Slavkovic-Lukic, Jessica Fioravanti, Azucena Martín-Santos & Luca Gattinoni

  2. IsoPlexis Corporation, Branford, CT, USA

    Edward Han & Jing Zhou

  3. Center for Immunomedicine in Transplantation and Oncology (CITO), University Hospital Regensburg, Regensburg, Germany

    Luca Gattinoni

  4. University of Regensburg, Regensburg, Germany

    Luca Gattinoni

Authors
  1. Dragana Slavkovic-Lukic
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  2. Jessica Fioravanti
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  3. Azucena Martín-Santos
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  4. Edward Han
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  5. Jing Zhou
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  6. Luca Gattinoni
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Corresponding authors

Correspondence to Dragana Slavkovic-Lukic or Luca Gattinoni .

Editor information

Editors and Affiliations

  1. Synthetic and Systems Biology for Biomedicine, Isituto Italiano di Tecnologia-IIT, Napoli, Napoli, Italy

    Velia Siciliano

  2. Department of Chemical Engineering, Imperial College, London, UK

    Francesca Ceroni

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Slavkovic-Lukic, D., Fioravanti, J., Martín-Santos, A., Han, E., Zhou, J., Gattinoni, L. (2024). Rapid Screening of CAR T Cell Functional Improvement Strategies by Highly Multiplexed Single-Cell Secretomics. In: Siciliano, V., Ceroni, F. (eds) Cancer Immunotherapy. Methods in Molecular Biology, vol 2748. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3593-3_11

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  • DOI: https://doi.org/10.1007/978-1-0716-3593-3_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3592-6

  • Online ISBN: 978-1-0716-3593-3

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