697 Pulsed Electric Fields induces a stepwise activation of host anti-tumor immunity in patients with early-stage non-small cell lung cancer (NSCLC)

BackgroundEnhancing anti-tumor immunity is a foundational therapeutic strategy against cancer.1 The Aliya Pulsed Electric Fields (PEF) proprietary system has been shown to promote local and systemic anti-tumor immune activation in pre-clinical murine models through the activation of immunogenic cell death (ICD) and the release of damage associated molecular patterns (DAMPs) and tumor-specific antigens.2 3 Unlike apoptosis, often immunosuppressive, ICD mechanisms generate a potent immune response by releasing DAMPs that are recognized by and attract immune cells.4 ICD mechanisms can suppress cancer cell proliferation and migration.5 6 In this study we evaluated PEF’s capability for modulating anti-tumor immunity in patients with NSCLC.MethodsThe treat-and-resect INCITE ES study enrolled patients with suspected or confirmed NSCLC stage IA2-IB and no cancer treatment history within two years. This two-arm study included 34 patients in the treatment group and 8 patients in the control group. Treatment group subjects received AliyaTM PEF (GTI-00018 investigational device; Galvanize Therapeutics) after the diagnostic biopsy, whereas the control group only had the biopsy. Blood, serum, and tissue samples were collected pre- and post-PEF at specific timepoints (figure 1).ResultsIngenuity Pathway Analysis (IPA) of serum cytokines predicts that host innate immunity pathways that drive immune cell trafficking, differentiation and activation are activated in PEF-treated samples, including the Acute Phase Response, IL-6, JAK-STAT and Th17/IL-17 signaling (figure 2A).7–9 scRNA-Seq from this initial cohort of patients additionally shows that PEF-treated tumors have a significant increase in the proportion of plasma B cells, T cells and neutrophils, with neutrophils expressing genes indicative of functional activation (figure 2B).7 In samples from PEF-treated patients, IPA further predicts activation of ICD mechanisms, such as pyroptosis and HMGB1 signaling (figure 3A,B).4 10 After this initial innate immune response, IPA predicts activation of Th2 signaling, responsible for dampening acute inflammation and orchestrating adaptive anti-tumor immunity (figure 4A).11 Flow cytometry analysis further suggests activation of adaptive immunity, as PEF samples have higher levels of circulating B cells and effector memory T cells, and lower Tregs (figure 4B). Within the tumor microenvironment, PEF tumors show increased proportion of cytotoxic CD8+ T cells, plasma B cells and tumor