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Understanding the role of calcium-mediated cell death in high-frequency irreversible electroporationWasson, Elisa M.; Alinezhadbalalami, Nastaran; Brock, Rebecca M.; Allen, Irving C.; Verbridge, Scott S.; Davalos, Rafael V.
Bioelectrochemistry, 02/2020, Letnik: 131Journal Article
High-frequency irreversible electroporation (H-FIRE) is an emerging electroporation-based therapy used to ablate cancerous tissue. Treatment consists of delivering short, bipolar pulses (1–10μs) in a series of 80–100 bursts (1 burst/s, 100μs on-time). Reducing pulse duration leads to reduced treatment volumes compared to traditional IRE, therefore larger voltages must be applied to generate ablations comparable in size. We show that adjuvant calcium enhances ablation area in vitro for H-FIRE treatments of several pulse durations (1, 2, 5, 10μs). Furthermore, H-FIRE treatment using 10μs pulses delivered with 1mM CaCl2 results in cell death thresholds (771±129V/cm) comparable to IRE thresholds without calcium (698±103V/cm). Quantifying the reversible electroporation threshold revealed that CaCl2 enhances the permeabilization of cells compared to a NaCl control. Gene expression analysis determined that CaCl2 upregulates expression of eIFB5 and 60S ribosomal subunit genes while downregulating NOX1/4, leading to increased signaling in pathways that may cause necroptosis. The opposite was found for control treatment without CaCl2 suggesting cells experience an increase in pro survival signaling. Our study is the first to identify key genes and signaling pathways responsible for differences in cell response to H-FIRE treatment with and without calcium. •CaCl2 enhances ablation volume for H-FIRE treatment of varying pulse widths.•CaCl2 enhances permeabilization of cells compared to a NaCl control.•CaCl2 upregulates expression of eIFB5 and 60s ribosomal subunit genes.•CaCl2 treatment leads to increased signaling in pathways that may cause necroptosis.•Treatment without CaCl2 upregulates expression of NOX1/4 genes.•Treatment without CaCl2 leads to an increase in pro survival signaling.
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