Fig. 5: Caspase-1 inhibition enhances chemotherapeutic efficacy by normalizing myelopoiesis.

a UMAP projection of pseudotime trajectory analysis, to infer cellular differentiation status of myeloid-lineage hematopoietic progenitor cell clusters, based on their differential gene expression using the Monocle3 pipeline (n = 2 mice per group, 16,130 number of total cells analyzed). b UMAP illustrating the hematopoietic and myeloid progenitor cell populations in various treatment conditions. c Stacked bar plot representing the relative distribution or frequency of each progenitor cell cluster, identified in the myeloid-lineage hematopoietic progenitor cell partitioned immune population. d Box plot illustrating the relative distribution of GMP amongst different treatment group (n = 2). e Schematic diagram illustrating GC chemotherapy treatment with or without caspase-1 inhibition, and endpoint collection of bone marrow, blood and tumor for further downstream analyses. Created in BioRender. Wong, S. (2026) https://BioRender.com/b3e0oiw. Flow cytometry analysis illustrating the percentage of GMP (f) and neutrophils (g), in the bone marrow of tumor-bearing mice treated with vehicle [black], gemcitabine-cisplatin chemotherapy (Chemo; Gemcitabine: 120 mg /kg, Cisplatin: 6 mg /kg, VX-765: 60 mg /kg) [red], VX-765 caspase-1 inhibitor (Casp1i) + Chemo combination [teal], and Casp1i [blue], respectively (n = 7 mice per group). h Flow cytometry analysis illustrating the percentage of neutrophils in the peripheral blood of tumor-bearing mice treated with vehicle [black], gemcitabine-cisplatin chemotherapy (Chemo; Gemcitabine: 120 mg/kg, Cisplatin: 6 mg/kg, VX-765: 60 mg/kg) [red], VX-765 caspase-1 inhibitor (Casp1i) + Chemo combination [teal], and Casp1i [blue], respectively (n = 5 per group). i Flow cytometry analysis illustrating the absolute count of intratumoral CD11b⁺Ly6G⁺ neutrophils, in tumor-bearing mice treated with vehicle [black], gemcitabine-cisplatin (Chemo; Gemcitabine: 120 mg/kg, Cisplatin: 6 mg/kg, VX-765: 60 mg/kg) [red], VX-765 caspase-1 inhibitor (Casp1i) + Chemo combination [teal], and Casp1i [blue], respectively (n = 7 mice per group). j Normalized absolute count of CD62L⁺CD11b⁺Ly6G⁺ neutrophils into tumors of mice treated with vehicle [black], gemcitabine-cisplatin (Chemo; Gemcitabine: 120 mg/kg, Cisplatin: 6 mg/kg, VX-765: 60 mg/kg) [red], VX-765 caspase-1 inhibitor (Casp1i) + Chemo combination [cyan], and Casp1i [blue], respectively (n = 7 mice per group). Tumor growth kinetics in mice harboring G69 (Gemcitabine: 120 mg/kg, Cisplatin: 6 mg/kg, VX-765: 60 mg/kg) (k) or 4T1 (Gemcitabine: 30 mg/kg, Cisplatin: 6 mg/kg, VX-765: 60 mg/kg) (l) xenografts treated with vehicle [black], gemcitabine-cisplatin (Chemo) [red], VX-765 caspase-1 inhibitor (Casp1i) + Chemo combination [teal], and Casp1i [blue], respectively (n = 7 mice per group). Statistics: Ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test (f–j) and two-way ANOVA with Geisser-Greenhouse correction followed by Dunnett’s multiple comparison test (k, l) were performed. Error bars represent standard error of mean (s.e.m.) unless otherwise specified. Abbreviations: hematopoietic stem and progenitor cell (HSPC), lympho-myeloid progenitor (LMPP), granulocyte-monocyte progenitors (GMP), common lymphoid progenitors (CMP), megakaryocyte-erythrocyte progenitors (MEP), monocyte progenitors (MoP), dendritic cell progenitors (DcP), neutrophil progenitor (NeuP), pre-neutrophil (preNeu), Pre-Basophil/Mast progenitors (Pre-BMP), mast cell progenitors (MaP), basophil progenitors (BaP) and Megakaryocyte progenitors (MkP). Box plots show the minimum and maximum values, and the center line represents the mean. Bone Marrow, Peripheral Blood and Intra-Tumoral cartoon icons were Created in BioRender. Wong, S. (2026) https://BioRender.com/b3e0oiw. Source data are provided as a Source Data file.