Session: 703. Cellular Immunotherapies: Basic and Translational: Poster II
Hematology Disease Topics & Pathways:
Research, clinical trials, Biological therapies, Translational Research, Clinical Research, genomics, immune mechanism, Therapies, metabolism, Biological Processes, Natural Killer (NK) Cell Therapies
Bone marrow (BM) aspirates and trephine biopsies for immune GE profiling and spatially resolved proteomics (IO360® panel, n = 740 genes, and GeoMx® DSP; NanoString Technologies) were collected at baseline (BL) and on treatment (OT) from 15 patients receiving CIML-NK cells on study NCT #01898793. ssGSEA-based scores were computed using the IOBR package in R. Machine learning was used to select features that correlated with response. Cell phenotypes were evaluated by mass cytometry. NK cell function was assayed in either conventional (N-; 20% IMDM) or TME-aligned media. Multi-omics studies were performed by mass spectrometry. Cell trafficking to TME was measured in NSG mouse tumors by tracking fluorescent-labeled WU-NK-101.
Correlative studies showed that BL GE signatures reflecting heightened neutrophil infiltration, chemokine receptor expression, and cancer antigen presentation predicted response to CIML-NKs (AUROC = 0.875, 0.875 and 0.850, respectively). We computed an 8-gene signature score (RPTOR, IRF9, TLR1, NLRP3, CD7, IL6R, FUT4, IL1RN) that was significantly higher at BL in responders and that correlated with response duration (Fig. 1A). Immune deconvolution of GE data with CIBERSORT highlighted critical differences in TME cellular composition at BL in responders to CIML-NKs, including a higher inferred abundance of macrophages, gamma/delta T cells and activated dendritic cells (Fig. 1B). In OT samples from responders, glycolytic GE programs as well as purine/pyrimidine and amino acid (AA) metabolism were upregulated compared with BL; no TME modulation was observed in non-responders (Fig. 1C). Analysis of WU-NK-101 cells revealed a unique phenotype compared with cNK cells, including higher expression of activating receptors, CD25, CD69, and GZMB, which positions WU-NK-101 for rapid activation and improved cytotoxicity. The latter hypothesis was confirmed by in vitro assays, with higher half maximal effective concentration (mEC50) for cNK vs WU-NK-101 (mEC50 = 5.2 vs 1.7; P = 0.032; Fig. 1D). The adverse impact of immunosuppressive TME-aligned media on cytotoxicity was evident on cNK but negligible to WU-NK-101 (mEC50 = 16.3 vs 2.0; P = 0.018; Fig. 1D). In N-media, WU-NK-101 used glucose as its main nutritional source; in hypoglycemic TME-aligned media, AA metabolic pathways were upregulated, indicating metabolic adaptability. This was supported by increased cell surface expression of AA transporters. WU-NK-101’s homing to the BM averaged 16.5 ± 2.4%, which is 5.5-fold higher than previously reported for feeder-cell+IL-2-expanded NK cells (~3%; Sato et al. Clin. Cancer Res. 2020).
In summary, an 8-gene TME signature showed excellent predictive ability for response to ML-NKs. In responders, ML-NK infusion was associated with TME modulation and with metabolic re-programming. Compared to cNK cells, WU-NK-101 had enhanced trafficking to the BM, anti-tumor activity, and a metabolic profile consistent with aerobic glycolysis, potentially contributing to mitigating TME adversity. Due to their resilience to an immunosuppressive TME, WU-NK-101 cells may represent an effective treatment modality for R/R AML; a phase 1 study of WU-NK-101 in R/R AML is in development (NCT #05470140).
Disclosures: Rutella: NanoString Technologies: Research Funding; MacroGenics: Research Funding; Wugen: Research Funding. Muth: Wugen: Current Employment, Current holder of stock options in a privately-held company. Mathyer: Wugen: Current Employment, Current holder of stock options in a privately-held company. Carter: Wugen, Inc.: Current Employment, Current holder of stock options in a privately-held company. Tumala: Wugen, Inc.: Current Employment, Current holder of stock options in a privately-held company. Arthur: Wugen: Current Employment, Current holder of stock options in a privately-held company. Magee: Wugen, Inc.: Current Employment, Current holder of stock options in a privately-held company. Petit: METAFORA biosystems: Current Employment, Current equity holder in private company. Blanchard: Exelixis: Current equity holder in publicly-traded company; Gilead: Current equity holder in publicly-traded company; Amgen: Current equity holder in publicly-traded company. Bhatnagar: Metafora Biosystems: Current Employment. Cooper: Wugen: Current Employment, Current holder of stock options in a privately-held company. Berrien-Elliott: Wugen: Consultancy, Current holder of stock options in a privately-held company, Patents & Royalties. Davidson-Moncada: Wugen: Current Employment, Current holder of stock options in a privately-held company. Fehniger: Indapta: Current holder of stock options in a privately-held company, Other: Scientific Advisory Board; Orca Bio: Current holder of stock options in a privately-held company; HCW Biologics: Research Funding; ImmunityBio: Research Funding; BMS: Other: Provision of drug for investigator initiated trial; Affimed: Other: Scientific Advisory Board, Research Funding; Wugen: Consultancy, Current holder of stock options in a privately-held company, Patents & Royalties.
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