Type: Oral
Session: 703. Cellular Immunotherapies other than CAR-T Cells: Basic and Translational: Enhancing NK Cell Therapeutics
Hematology Disease Topics & Pathways:
Research, Translational Research
Using flow cytometry, we confirmed that W-NK1 drug product has higher expression of NKG2A, 2B4, and DNAM-1 as well as the immune engaging receptors CD100, CD86 and CD337 compared to donor-matched cNK cells. Upon activation by target cell killing, W-NK1 cells release a variety of cytokines and chemokines which lead to maturation and activation of dendritic cells (DC) (TNFa, IFNg, MIP-1b/CCL4, RANTES/CCL5, MIP-3a/CCL20), migration and activation of T cells (CXCL9, CXCL10, CXCL11), and maturation of macrophages (CCL2, CCL5). Indeed, we show that co-culture of immature DC, with W-NK1 in the presence of AML cells caused increased expression of the activation markers CD40, CD80, CD83 and CD86 leading to enhanced phagocytosis of target cells (~3-fold greater than when incubated with cNK). Additionally, W-NK1 secreted chemokines have the ability to recruit T-cells in a transwell assay, suggesting that W-NK1 are able to recruit T-cells into the TME. In mixed lymphocyte reactions combining NK with PBMCs and donor matched monocyte-derived DC, T cell activation (determined by CD25 expression) and proliferation was significantly enhanced when W-NK1 was compared to cNK (P <0.01). In the absence of matured DC T-cell proliferation was much lower (P <0.01). Overall, this demonstrated coordination between NK cells and DC toward T-cell activation that was significantly enhanced by W-NK1. Finally, we assessed the impact of W-NK1 on monocyte-derived M1 vs. M2 macrophages in a co-culture assay. W-NK1 demonstrated selective cytotoxicity against pro-tumorigenic M2 macrophages while sparing M1 macrophages (P < 0.001) indicating an ability to tilt the TME toward a less immunosuppressive environment.
These findings were confirmed in multiple humanized AML xenograft NCG models (THP-1 and HL-60). In vivo, W-NK1 synergized with the human immune system to coordinate robust anti-AML responses. In concert with human immune cells, W-NK1 treatment led to deeper (>10-fold) and more durable anti-tumor responses compared to humanization or W-NK1 treatment alone (P <0.05). Immune engagement was confirmed by immunofluorescence staining of the tumor samples on a GeoMx Digital Spatial Profiling platform (NanoString Technologies, San Diego, CA). Furthermore, splenocyte-derived T-cells extracted from HL-60 humanized NCG mice treated with W-NK1-demonstrated a significantly enhanced ability to kill the parental HL-60 tumor cells ex vivo (~2-fold vs. vehicle at E:T 3:1; P <0.001). Importantly, the splenocyte-derived T-cells appear to retain specificity as T-cells ‘educated’ against the HL-60 tumor cell line were less effective against an alternate THP-1 AML tumor type (1.0-fold vs. vehicle at E:T 3:1) indicating potentiation of allo-response.
In summary, our studies demonstrate that the unique properties of W-NK1 leverages potent engagement with the endogenous immune system through multiple mechanisms. This exquisite synergy between innate and adaptive mechanisms is enabled through W-NK1 reconditioning of the adaptive immune response, effectively enhancing recruitment, proliferation, activation state and antigen presentation leading to a deep and durable adaptive response. In this manner the pharmacodynamic effect of W-NK1 extends far beyond the pharmacokinetic effect. These data augur positively for patients to be treated with W-NK1 in future studies.
Disclosures: Leedom: Wugen: Current Employment, Current holder of stock options in a privately-held company. Muz: Wugen: Current Employment, Current holder of stock options in a privately-held company. Magee: Wugen: Current Employment, Current holder of stock options in a privately-held company. Vadakekolathu: Wugen: Research Funding. Arthur: Wugen: Current Employment, Current holder of stock options in a privately-held company. Tran: Wugen: Current Employment, Current holder of stock options in a privately-held company. Luukkonen: Wugen: Current Employment, Current holder of stock options in a privately-held company. Mahajan: Wugen: Current Employment, Current holder of stock options in a privately-held company. Hamil: Wugen: Current Employment, Current holder of stock options in a privately-held company. Muth: Wugen: Current Employment, Current holder of stock options in a privately-held company. Berrien-Elliott: Wugen: Consultancy, Current equity holder in private company. Fehniger: Affimed: Other: Scientific Advisory Board; Wugen: Consultancy, Current holder of stock options in a privately-held company, Research Funding; Orca Bio: Current holder of stock options in a privately-held company; Indapta: Current holder of stock options in a privately-held company; Smart Immune: Other: Scientific Advisory Board; AI Proteins: Other: Scientific Advisory Board, Research Funding. Rutella: Wugen: Research Funding. Davidson-Moncada: Wugen: Current Employment, Current holder of stock options in a privately-held company.