Continuously expanding CAR NK-92 cells display selective cytotoxicity against B-cell leukemia and lymphoma

Cytotherapy. 2017 Feb;19(2):235-249. doi: 10.1016/j.jcyt.2016.10.009. Epub 2016 Nov 22.

Abstract

Background aims: Natural killer (NK) cells can rapidly respond to transformed and stressed cells and represent an important effector cell type for adoptive immunotherapy. In addition to donor-derived primary NK cells, continuously expanding cytotoxic cell lines such as NK-92 are being developed for clinical applications.

Methods: To enhance their therapeutic utility for the treatment of B-cell malignancies, we engineered NK-92 cells by lentiviral gene transfer to express chimeric antigen receptors (CARs) that target CD19 and contain human CD3ζ (CAR 63.z), composite CD28-CD3ζ or CD137-CD3ζ signaling domains (CARs 63.28.z and 63.137.z).

Results: Exposure of CD19-positive targets to CAR NK-92 cells resulted in formation of conjugates between NK and cancer cells, NK-cell degranulation and selective cytotoxicity toward established B-cell leukemia and lymphoma cells. Likewise, the CAR NK cells displayed targeted cell killing of primary pre-B-ALL blasts that were resistant to parental NK-92. Although all three CAR NK-92 cell variants were functionally active, NK-92/63.137.z cells were less effective than NK-92/63.z and NK-92/63.28.z in cell killing and cytokine production, pointing to differential effects of the costimulatory CD28 and CD137 domains. In a Raji B-cell lymphoma model in NOD-SCID IL2R γnull mice, treatment with NK-92/63.z cells, but not parental NK-92 cells, inhibited disease progression, indicating that selective cytotoxicity was retained in vivo.

Conclusions: Our data demonstrate that it is feasible to generate CAR-engineered NK-92 cells with potent and selective antitumor activity. These cells may become clinically useful as a continuously expandable off-the-shelf cell therapeutic agent.

Keywords: B-cell malignancies; CD19; adoptive immunotherapy; chimeric antigen receptor; natural killer cells.

MeSH terms

  • Animals
  • Antigens, CD19 / immunology
  • B-Lymphocytes / immunology
  • CD3 Complex / genetics
  • CD3 Complex / metabolism
  • Cell Line, Tumor
  • Cell Proliferation
  • Cytotoxicity, Immunologic* / genetics
  • Epitopes / genetics
  • HEK293 Cells
  • Humans
  • Immunotherapy, Adoptive / methods
  • Killer Cells, Natural* / immunology
  • Killer Cells, Natural* / metabolism
  • Killer Cells, Natural* / transplantation
  • Leukemia, Lymphocytic, Chronic, B-Cell / immunology
  • Leukemia, Lymphocytic, Chronic, B-Cell / therapy*
  • Lymphocyte Activation / genetics
  • Lymphoma / immunology
  • Lymphoma / therapy*
  • Lymphoma, B-Cell / immunology
  • Lymphoma, B-Cell / therapy
  • Male
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / metabolism*
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / genetics
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / metabolism

Substances

  • Antigens, CD19
  • CD3 Complex
  • CD3 antigen, zeta chain
  • Epitopes
  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor Receptor Superfamily, Member 9