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Heparanase promotes tumor infiltration and antitumor activity of CAR-redirected T lymphocytes

Nature Medicine volume 21, pages 524–529 (2015)Cite this article

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Abstract

Adoptive transfer of chimeric antigen receptor (CAR)-redirected T lymphocytes (CAR-T cells) has had less striking therapeutic effects in solid tumors1,2,3 than in lymphoid malignancies4,5. Although active tumor-mediated immunosuppression may have a role in limiting the efficacy of CAR-T cells6, functional changes in T lymphocytes after their ex vivo manipulation may also account for the reduced ability of cultured CAR-T cells to penetrate stroma-rich solid tumors compared with lymphoid tissues. We therefore studied the capacity of human in vitro–cultured CAR-T cells to degrade components of the extracellular matrix (ECM). In contrast to freshly isolated T lymphocytes, we found that in vitro–cultured T lymphocytes lack expression of the enzyme heparanase (HPSE), which degrades heparan sulfate proteoglycans, the main components of ECM. We found that HPSE mRNA is downregulated in in vitro–expanded T cells, which may be a consequence of p53 (officially known as TP53, encoding tumor protein 53) binding to the HPSE gene promoter. We therefore engineered CAR-T cells to express HPSE and showed their improved capacity to degrade the ECM, which promoted tumor T cell infiltration and antitumor activity. The use of this strategy may enhance the activity of CAR-T cells in individuals with stroma-rich solid tumors.

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Figure 1: LTE-T cells show reduced invasion of the ECM and loss of the enzyme HPSE.
Figure 2: LTE-T cells modified to express HPSE show enhanced degradation of the ECM.
Figure 3: LTE-T cells co-expressing HPSE and GD2-specific CAR retain GD2 specificity and have enhanced capacity to degrade ECM.
Figure 4: CAR-GD2+ LTE-T cells co-expressing HPSE show enhanced tumor infiltration and improve overall survival in xenograft tumor models.

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Acknowledgements

The authors would like to thank I.Vlodavsky and M. Brenner for the critical review of the manuscript and C. Gillespie for editing. This work was supported in part by the US National Institutes of Health-National Cancer Institute (G.D., no. R01 CA142636) and by a Department of Defense and Technology and Therapeutic Development Award (G.D., no. W81XWH-10-10425). L. Metelitsa kindly provided the CHLA-255 human NB cell line.

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Authors and Affiliations

  1. Center for Cell and Gene Therapy, Baylor College of Medicine and Houston Methodist Hospital, Houston, Texas, USA

    Ignazio Caruana, Barbara Savoldo, Valentina Hoyos, Gerrit Weber & Gianpietro Dotti

  2. Department of Pediatrics, Texas Children's Hospital, Baylor College of Medicine, Houston, Texas, USA

    Barbara Savoldo

  3. Biostatistics Shared Resource, Baylor College of Medicine, Houston, Texas, USA

    Hao Liu

  4. Department of Surgery, Texas Children's Hospital, Baylor College of Medicine, Houston, Texas, USA

    Eugene S Kim

  5. Department of Pathology and Immunology, Baylor College of Medicine, Houston, Texas, USA

    Michael M Ittmann, Dario Marchetti & Gianpietro Dotti

  6. Interdepartmental Program in Translational Biology and Molecular Medicine, Baylor College of Medicine, Houston, Texas, USA

    Michael M Ittmann

  7. Michael E. DeBakey Department of Veterans Affairs Medical Center, Dan L. Duncan Cancer Center, Houston, Texas, USA

    Michael M Ittmann

  8. Department of Medicine, Baylor College of Medicine, Houston, Texas, USA

    Gianpietro Dotti

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Contributions

G.D., I.C. and B.S. designed experiments; I.C., V.H., G.W. and E.S.K. performed the experiments; I.C., B.S. and G.D. analyzed the data; I.C. and G.D. wrote the manuscript; H.L. performed the statistical analysis; M.M.I. performed the pathology; D.M. provided his expertise in the heparanase field and provided crucial reagents; all the authors reviewed and approved the final version of the manuscript.

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Correspondence to Gianpietro Dotti.

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Competing interests

G.D. and B.S. have ownership interest (including patents) in the field of T cell and gene-modified T cell therapy for cancer and have a collaborative research agreement with Celgene and Bluebird Bio.

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Caruana, I., Savoldo, B., Hoyos, V. et al. Heparanase promotes tumor infiltration and antitumor activity of CAR-redirected T lymphocytes. Nat Med 21, 524–529 (2015). https://doi.org/10.1038/nm.3833

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