Inhibition of WEE1 kinase and cell cycle checkpoint activation sensitizes head and neck cancers to natural killer cell therapies

Abstract

BackgroundNatural killer (NK) cells recognize and lyse target tumor cells in an MHC-unrestricted fashion and complement antigen- and MHC-restricted killing by T-lymphocytes. NK cells and T-lymphocytes mediate early killing of targets through a common granzyme B-dependent mechanism. Tumor cell resistance to granzyme B and how this alters NK cell killing is not clearly defined.MethodsTumor cell sensitivity to cultured murine KIL and human high affinity NK (haNK) cells in the presence or absence of AZD1775, a small molecule inhibitor of WEE1 kinase, was assessed via real time impedance analysis. Mechanisms of enhanced sensitivity to NK lysis were determined and in vivo validation via adoptive transfer of KIL cells into syngeneic mice was performed.ResultsCultured murine KIL cells lyse murine oral cancer 2 (MOC2) cell targets more efficiently than freshly isolated peripheral murine NK cells. MOC2 sensitivity to granzyme B-dependent KIL cell lysis was enhanced by inhibition of WEE1 kinase, reversing G2/M cell cycle checkpoint activation and resulting in enhanced DNA damage and apoptosis. Treatment of MOC2 tumor-bearing wild-type C57BL/6 mice with AZD1775 and adoptively transferred KIL cells resulted in enhanced tumor growth control and survival over controls or either treatment alone. Validating these findings in human models, WEE1 kinase inhibition sensitized two human head and neck cancer cell lines to direct lysis by haNK cells. Further, WEE1 kinase inhibition sensitized these cell lines to antibody-dependent cell-mediated cytotoxicity when combined with the anti-PD-L1 IgG1 mAb Avelumab.ConclusionsTumor cell resistance to granzyme B-induced cell death can be reversed through inhibition of WEE1 kinase as AZD1775 sensitized both murine and human head and neck cancer cells to NK lysis. These data provide the pre-clinical rationale for the combination of small molecules that reverse cell cycle checkpoint activation and NK cellular therapies.

Tumor control via targeting PD-L1 with chimeric antigen receptor modified NK cells

“…To overcome this requirement, haNKs, or high-affinity NK cells, are NK-92 cells engineered to express endoplasmic reticulum-retained IL-2(11). haNKs have demonstrated the ability to efficiently kill carcinoma cells at low effector-to-target ratios(12, 13). Irradiation of haNKs and haNK-derived cell products ensures short life span of these cells and allows for repeat administration with no risk of cellular engraftment and low risk of cytopenias in treated hosts(14, 27).…”

“…NK-92 cells, derived from a Non-Hodgkin’s lymphoma patient, can be continuously expanded in culture and following irradiation, can be safely administered in high doses to patients with cancer(8-10). High-affinity NK (haNK) cells are NK-92 cells engineered to express endoplasmic reticulum-retained IL-2(11), have demonstrated potent effector function in numerous pre-clinical models(12, 13), and following irradiation, can also be safely administered in high doses to patients(14). haNKs represent an “off the shelf” NK cellular therapy available for pre-clinical and clinical study.…”

Unleashing Natural Killer Cells in the Tumor Microenvironment–The Next Generation of Immunotherapy?

“…Friedman et al recently utilized an inhibitor for the WEE1 kinase (AZD1775) on head and neck cancer cells, which reversed G2/M cell cycle checkpoint activation of the cancer cells, resulting in increased DNA damage and susceptibility to NK cell-mediated killing (318). Interestingly, DNA cell cycle checkpoint in this study did not impact surface expression of MHC-I, PD-L1, or NKG2D ligands (RAE, H60, MULT-1) on cancer cells, despite increasing their susceptibility to NK cells.…”

Scientifically based combination therapies with immuno‐oncology checkpoint inhibitors

“…This study also reported evidence of immunologic memory after combination treatment 172 . AZD1775 also sensitises human head and neck cancer cells to avelumab, improving antibody‐dependent cellular cytotoxicity‐mediated killing of these cancer cells 173 . AZD1775 is being trialled as a monotherapy in various tumour types such as uterine cancer (NCT03668340) and relapsed small cell lung cancer (NCT02593019).…”