Gene-modified NK-92MI cells expressing a chimeric CD16-BB-ζ or CD64-BB-ζ receptor exhibit enhanced cancer-killing ability in combination with therapeutic antibody

Abstract

Natural killer (NK) cells play a pivotal role in monoclonal antibody-mediated immunotherapy through the antibody-dependent cell-mediated cytotoxicity (ADCC) mechanism. NK-92MI is an interleukin-2 (IL-2)-independent cell line, which was derived from NK-92 cells with superior cytotoxicity toward a wide range of tumor cells in vitro and in vivo. Nonetheless, the Fc-receptor (CD16) that usually mediates ADCC is absent in NK-92 and NK-92MI cells. To apply NK-92MI cell-based immunotherapy to cancer treatment, we designed and generated two chimeric receptors in NK-92MI cells that can bind the Fc portion of human immunoglobulins. The construct includes the low-affinity Fc receptor CD16 (158F) or the high-affinity Fc receptor CD64, with the addition of the CD8a extracellular domain, CD28 transmembrane domains, two costimulatory domains (CD28 and 4-1BB), and the signaling domain from CD3ζ. The resulting chimeric receptors, termed CD16-BB-ζ and CD64-BB-ζ, were used to generate modified NK-92MI cells expressing the chimeric receptor, which were named NK-92MIhCD16 and NK-92MIhCD64 cells, respectively. We found that NK-92MIhCD16 and NK-92MIhCD64 cells significantly improved cytotoxicity against CD20-positive non-Hodgkin's lymphoma cells in the presence of rituximab. These results suggest that the chimeric receptor-expressing NK-92MI cells may enhance the clinical responses to currently available anticancer monoclonal antibodies.

Overcoming Resistance to Natural Killer Cell Based Immunotherapies for Solid Tumors

“…The insertion of the high affinity CD16 FcγRIIIa (158V) allele and IL-2 into NK-92 cells render NK-mediated ADCC using cetuximab, trastuzumab and pertuzumab against a variety of solid tumor cells (165). Additional strategies include engineering NK cells with chimeric receptors CD16-BB-ζ and CD64-BB-ζ (166). These engineered NK cells significantly improved cytotoxicity against CD20-positive NHL cells in the presence of rituximab (166) but their anti-tumor effects need to be evaluated for solid tumor cells with targeted antibodies.…”

“…Additional strategies include engineering NK cells with chimeric receptors CD16-BB-ζ and CD64-BB-ζ (166). These engineered NK cells significantly improved cytotoxicity against CD20-positive NHL cells in the presence of rituximab (166) but their anti-tumor effects need to be evaluated for solid tumor cells with targeted antibodies.…”

Chimeric Antigen Receptor Expressing Natural Killer Cells for the Immunotherapy of Cancer

“…Indeed, in a study of NK-92 cells engineered with ErbB2/HER2-CAR, while irradiation had no effect on the in vitro cytotoxicity of CAR-transduced NK92 cells, it negatively impacted their in vivo replication and persistence, with the cells no longer detectable within 7 days of adoptive infusion ( 109 ). Of note, NK-92 cells are CD16 (FCRIIIγ) negative and cannot mediate antibody-dependent cell cytotoxicity (ADCC), unless genetically modified to express CD16 ( 120 ).…”

Nano-loaded natural killer cells as carriers of indocyanine green for synergetic cancer immunotherapy and phototherapy

“…[18][19][20] In particular, NK92 cells derived from NK lymphoma are considered to have high cytotoxicity over malignant cells, to be relatively long-lived when expanded ex vivo, and to be safe and more frequently used in clinical studies. 21 In this study, an NK92-derived and highly toxic interleukin-2-independent cell line, 22 NK-92MI cells were used in order to recognize and target the cancer cell antigens along cellular surface for cancer therapy. 23 NK-92MI cells can secrete perforin and granzymes that are normally stored inside the secretory lysosomes.…”