Abstract

T cell malignancies represent a group of hematologic cancers with high rates of relapse and mortality in patients for whom no effective targeted therapies exist. The shared expression of target antigens between chimeric antigen receptor (CAR) T cells and malignant T cells has limited the development of CAR-T because of unintended CAR-T fratricide and an inability to harvest sufficient autologous T cells. Here, we describe a fratricide-resistant “off-the-shelf” CAR-T (or UCART7) that targets CD7+ T cell malignancies and, through CRISPR/Cas9 gene editing, lacks both CD7 and T cell receptor alpha chain (TRAC) expression. UCART7 demonstrates efficacy against human T cell acute lymphoblastic leukemia (T-ALL) cell lines and primary T-ALL in vitro and in vivo without the induction of xenogeneic GvHD. Fratricide-resistant, allo-tolerant “off-the-shelf” CAR-T represents a strategy for treatment of relapsed and refractory T-ALL and non-Hodgkin’s T cell lymphoma without a requirement for autologous T cells.

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Acknowledgements

This work is dedicated in memory of Gordon S. Cooper. 1946–2017. We thank Dr. Carl June (University of Pennsylvania) for providing the backbone of a third-generation CAR and the pELNS-Ef1α lentiviral vector.

Funding

Specialized Program of Research Excellence (SPORE) in Leukemia NIH: 1P50CA171063-01A1, R35 CA210084-01A, the Gabrielle’s Angels Foundation, the Children's Discovery Institute of Washington University and St. Louis Children's Hospital, the Alvin J. Siteman Cancer Research Fund at Washington University in St. Louis, MO.

Author contributions

MLC and JFD conceived project. MLC, JFD, JC, and MR designed the experiments. MLC, JKR, JMN, and KE cloned the CAR constructs and generated virus. MLC performed gene editing and generated CAR-T. MLC, JKR, and JO preformed and analyzed in vitro assays. MLC, JKR, JMN, BW, and LNG performed in vivo experiments. JLP and SA performed BLI imaging. KS and MLC performed FACS analysis. DMW and AG developed PDX models. MLC, CAM, CCF, and RSF completed and analyzed off-target nuclease activity analysis. FG performed all statistical analyses. All authors were involved in the interpretation of data and preparation of this manuscript.

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Affiliations

  1. Department of Internal Medicine, Division of Oncology, Washington University School of Medicine, St. Louis, MO, 63110, USA

    • Matthew L Cooper
    • , Jaebok Choi
    • , Karl Staser
    • , Julie K Ritchey
    • , Jessica M Devenport
    • , Kayla Eckardt
    • , Michael P Rettig
    • , Bing Wang
    • , Linda G Eissenberg
    • , Armin Ghobadi
    • , Leah N Gehrs
    • , Christopher A Miller
    • , Julie O’Neal
    •  & John F DiPersio
  2. Department of Internal Medicine, Division of Dermatology, Washington University School of Medicine, St. Louis, MO, 63110, USA

    • Karl Staser
  3. Mallinckrodt Institute of Radiology, Washington University in St. Louis School of Medicine, St. Louis, MO, 63110, USA

    • Julie L Prior
    •  & Samuel Achilefu
  4. McDonnell Genome Institute, Washington University School of Medicine, St. Louis, MO, 63108, USA

    • Christopher A Miller
    • , Catrina C Fronick
    •  & Robert S Fulton
  5. Department of Surgery, Division of Public Health Sciences, Washington University School of Medicine, St. Louis, MO, 63110, USA

    • Feng Gao
  6. Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA

    • David M Weinstock
    •  & Alejandro Gutierrez
  7. Division of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, 02215, USA

    • Alejandro Gutierrez

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The authors declare that they have no conflict of interest.

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Correspondence to Matthew L Cooper or John F DiPersio.

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DOI

https://doi.org/10.1038/s41375-018-0065-5

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