Checking a set of hypotheses

The intervention uses noninvasive intermediate-frequency alternating electric fields, termed Tumor Treating Fields (TTFields), at 150-200 kHz.

Matches this study. The study uses noninvasive alternating electric fields and applies 200 kHz to glioblastoma cells, within the specified range. Its 150 kHz experiments concern lung and pancreatic cancer cells.

Tumor Treating Fields dually activate STING and AIM2 inflammasomes to induce adjuvant immunity in glioblastoma (Abstract; Results, Figure 1A and accompanying text; Supplemental Figure 4 description.)

TTFields cause focal nuclear-envelope disruption and the formation of cytosolic micronuclear clusters in dividing glioblastoma cells.

Matches this study. Microscopy demonstrates focal nuclear-envelope rupture and cytosolic micronuclear clusters in glioblastoma cells. Ribociclib experiments establish dependence on cell-cycle entry; affected cells need not be in mitosis.

Tumor Treating Fields dually activate STING and AIM2 inflammasomes to induce adjuvant immunity in glioblastoma (Results, Figure 1A-D; Supplemental Figures 1-7 descriptions.)

Nuclear-envelope disruption and cytosolic micronuclear clusters recruit and activate the cytosolic DNA sensors cGAS/STING and AIM2/caspase-1 inflammasomes.

Matches this study. The study demonstrates sensor recruitment to micronuclear clusters, STING-dependent inflammatory signaling, and AIM2-dependent caspase-1 activation, supported by depletion and rescue experiments.

Tumor Treating Fields dually activate STING and AIM2 inflammasomes to induce adjuvant immunity in glioblastoma (Results, Figures 1-5; STING and AIM2 depletion and rescue experiments.)

This signaling converts the immunosuppressive tumor microenvironment into an immunopermissive state, with activation of dendritic cells and adaptive CD8+ T-cell immunity.

Matches this study. Conditioned-medium and syngeneic implantation experiments demonstrate dendritic-cell activation, activated CD8+ T cells, tumor immune infiltration, and memory responses dependent on tumor-cell STING/AIM2 expression. These animal results follow implantation of cells pretreated with TTFields in vitro.

Tumor Treating Fields dually activate STING and AIM2 inflammasomes to induce adjuvant immunity in glioblastoma (Results, Figures 6-10; Figures 7E-F and 8D-E.)

The proposed TTFields-induced mechanism increases survival in patients with glioblastoma receiving maintenance temozolomide.

Not verified in the article. The 12-patient cohort measures immune changes after initiation of TTFields and maintenance temozolomide, rather than testing whether this mechanism mediates improved survival. The survival trial is cited, but its full article is absent from the supplied sources, and the causal mechanism-to-survival link remains unverified.

Tumor Treating Fields dually activate STING and AIM2 inflammasomes to induce adjuvant immunity in glioblastoma (Results, Figures 11-15; Discussion; reference 81.)

The decisive experiment uses the syngeneic KR158-luc glioblastoma model.

Matches this study. Luciferase-tagged KR158 cells are used in an orthotopic glioblastoma model syngeneic to C57BL/6J mice, matching the specified model.

Tumor Treating Fields dually activate STING and AIM2 inflammasomes to induce adjuvant immunity in glioblastoma (Results, Figures 6, 7 and 9; Table 1.)

The decisive experiment is specified as a 2×2 design comparing wild type with STING/AIM2 double knockout (DKO), crossed with TTFields versus a sham-heat thermal control and temozolomide monotherapy; the placement of temozolomide monotherapy within the stated 2×2 design remains unspecified.

Different experimental conditions. The four groups cross scrambled-control versus dual STING/AIM2 knockdown with untreated versus TTFields-pretreated cells. Knockdown substitutes for the specified knockout, and a sham-heat control is not verified. Temozolomide alone and with TTFields is tested separately using 300 μM for 72 hours before implantation, rather than within the specified knockout factorial experiment. The proposed placement of temozolomide within 2×2 remains ambiguous.

Tumor Treating Fields dually activate STING and AIM2 inflammasomes to induce adjuvant immunity in glioblastoma (Results, Figure 7A-D; Supplemental Figure 15 description; Methods.)

The biological target is the voltage-gated sodium channel NaV1.5, with SCN5A as its canonical target identifier, aberrantly expressed in human breast cancer cells.

Matches this study. Driffort and colleagues studied functional NaV1.5 in human MDA-MB-231 breast cancer cells, measured SCN5A expression, and suppressed its expression with targeted shRNA.

Ranolazine inhibits NaV1.5-mediated breast cancer cell invasiveness and lung colonization (Findings; Figures 1 and 2A-B.)

The intervention uses ranolazine, described as an approved phase 4 drug, at 50 µM in vitro and 50 mg/kg/day in vivo.

Different experimental conditions. The study used 50 µM ranolazine in vitro and 50 mg/kg intraperitoneally in mice, but administered the animal treatment five days per week. It identifies ranolazine as approved for chronic angina; the phase 4 designation is not documented.

Ranolazine inhibits NaV1.5-mediated breast cancer cell invasiveness and lung colonization (Findings, ranolazine pharmacology and animal experiment; Figures 1, 2E-G and 3A.)

Ranolazine selectively inhibits the persistent sodium current of NaV1.5, thereby eliminating pathological membrane depolarization and suppressing proteolytic matrix degradation.

Not verified in the article. Ranolazine inhibited sodium currents and reduced focal matrix proteolysis, with an invasion assay supporting NaV1.5 dependence. However, the reported electrophysiology measured peak currents and gating; it did not establish selective persistent-current inhibition followed by elimination of pathological membrane depolarization as the cause of reduced proteolysis.

Ranolazine inhibits NaV1.5-mediated breast cancer cell invasiveness and lung colonization (Findings; Figure 1A-C and Figure 2E-G.)

Ranolazine treatment is predicted to reduce metastatic lung colonization without cardiotoxicity.

Not verified in the article. Ranolazine reduced the lung bioluminescence signal by 77 ± 8% relative to control, supporting reduced colonization burden. Similar body-weight trajectories and the authors' description of no apparent toxicity do not establish absence of cardiotoxicity; cardiac-specific measurements are not reported.

Ranolazine inhibits NaV1.5-mediated breast cancer cell invasiveness and lung colonization (Findings, lung colonization results; Figure 3B-E; concluding discussion.)

The decisive experiment uses NMRI nude mice bearing orthotopic MDA-MB-231 xenografts.

Different experimental conditions. The ranolazine study used NMRI nude mice and MDA-MB-231-Luc cells, but injected cells into the tail vein rather than establishing orthotopic xenografts. Its three groups contained 18, 12 and 8 mice, totaling 38. Nelson and colleagues separately studied orthotopic MDA-MB-231 tumors in Rag2−/− Il2rg−/− mice.

Ranolazine inhibits NaV1.5-mediated breast cancer cell invasiveness and lung colonization (Findings, animal experiment; Figure 3A-E.)

The decisive comparison includes four groups: control, ranolazine, SCN5A-targeting short hairpin RNA (shRNA), and SCN5A-targeting shRNA combined with ranolazine.

Different experimental conditions. The animal experiment compared control cells, SCN5A-suppressed cells, and control cells with ranolazine; it omitted the combined SCN5A-suppression plus ranolazine group. That combination was examined in vitro, where ranolazine produced no further reduction of invasion after NaV1.5 suppression.

Ranolazine inhibits NaV1.5-mediated breast cancer cell invasiveness and lung colonization (Figure 2E; Findings, animal experiment; Figure 3A-C.)

The experiment includes control of mitochondrial β-oxidation, with the form of that control left unspecified.

Not verified in the article. No supplied study documents control of mitochondrial β-oxidation within the proposed ranolazine experiment. The separate metabolic study measured glycolysis and mitochondrial respiration under other interventions, which does not verify this β-oxidation control.

The experiment includes electrocardiographic monitoring of the QT interval.

Not verified in the article. No supplied study reports electrocardiographic QT monitoring in the proposed breast cancer experiment.