Extended Data Fig. 11: Immune cell dependence of STING agonist-mediated suppression of metastasis reactivation.
From: STING inhibits the reactivation of dormant metastasis in lung adenocarcinoma
(a) Rationale for experimental design. (b-c) qRT-PCR analysis of CXCL10 and CCL5 mRNA levels in WT or STING knockout KPad1 (b) or H2087-LCC (c) cells treated with 33 μM MSA-2 for 4 h. Mean ± s.e.m., representative of 2 independent experiments. Each dot represents a technical replicate of the assay. (d–e) Treatment was performed as in (b, c) but with 10 μM (KPad1) or 50 μM (H2087-LCC) ADU-S100. Mean ± s.e.m., representative of 2 independent experiments. Each dot represents a technical replicate of the assay. (f-j) Metastasis-free survival plots of B6-albino mice intracardially inoculated with 2.5x105 KPad1 cells, treated with both MSA-2 and IgG control (f, n = 10 mice) or individual antibodies to deplete NK cells (g, n = 10 mice for vehicle or 8 mice for MSA-2), CD4+ T cells (h, n = 10 mice), CD8+ T cells (i, n = 10 mice), or a combination of these antibodies (j, n = 10 mice for vehicle or 9 mice for MSA-2). Mice were administered antibodies (200 μg/mouse) once weekly for 3 weeks starting 6 days after cell inoculation, and vehicle or MSA-2 (50 mg/kg of body weight) once weekly for 2 weeks starting 9 days after cell inoculation. Log-rank test. (k) Metastasis-free survival plots of B6-albino mice intracardially inoculated with 2.5 x 105 KPad1 cells pre-treated with 33 μM MSA-2 for 24h before inoculation. n = 10 mice per group. (l) Metastasis-free survival plots of C57BL/6J mice intravenously inoculated with 2.5 x 105 WT or Sting knockout KPad1 cells and treated with vehicle or ADU-S100 (1.25 mg/kg of body weight) once weekly by intratracheal delivery until the study endpoint. ADU-S100 treatment started 5 days after cell inoculation. n = 15 mice (control) or 8 mice (ADU-S100). Log-rank test. (m) Quantification of WT or STING knockout H2087-LCC cell numbers in lungs from athymic mice treated with vehicle or ADU-S100 (6.25 mg/kg of body weight) once weekly for 4 weeks by intratracheal delivery. ADU-S100 treatment started 1 week after intravenous inoculation of 1 x 105 cells. Lungs were harvested 5 weeks after inoculation. n = 5 mice per group. Mean ± s.e.m., two-sided unpaired t-test.
