Journal: Cancer Research Communications
Article Title: Dipeptidyl Peptidase 4 Restoration Facilitates Antitumor Immunity in KRAS-LKB1–Mutant Lung Cancer
doi: 10.1158/2767-9764.CRC-25-0199
Figure Lengend Snippet: DPP4 is a potential therapeutic target in KRAS -mutant cells via NK cell recruitment. A, Immunoblotting of the indicated proteins in KL H2122 cells transduced with the indicated vectors. B, DPP4 activity induced by DPP4-Glo in KL H2122 cells transduced with the indicated vectors ( n = 3). C, GSEA showing significantly differentially expressed pathways. GSEA Gene Ontology Biological Process (GO BP) category upregulation in DPP4-reconstituted H2122 cells relative to luciferase (LUC)-reconstituted H2122 cells. D, GSEA of NK cell activation involved in the immune response signature in H2122 cells transduced with DPP4 or LUC. E, ELISA for human granzyme B in conditioned medium (CM) derived from H2122 cells cocultured with NK-92 cells ( n = 3). F, Schematic of an immune cell migration assay using a three-dimensional microfluidic device with tumor spheroids embedded in a central collagen-filled channel and immune cells cocultured in a side channel. G, Representative images of NK-92 cell migration toward H2122 or DPP4-overexpressing H2122 cells (left). Immune infiltration into the peritumor region was quantified using ImageJ software (right). Scale bars, 500 μm. H, Representative images of NK-92 cell migration in KP H2009 cells treated with and without sitagliptin (left). Immune infiltration into the peritumor region was quantified using ImageJ software (right). Scale bars, 500 μm. I, ELISA for human granzyme B in CM derived from H2009 cells treated with and without sitagliptin and cocultured with NK-92 cells ( n = 3). J, Evaluation of H2122-specific CD8 + T-cell migration toward H2122 or DPP4-overexpressing H2122 cells. Immune infiltration into the peritumor region was quantified using ImageJ software. K, ELISA for human granzyme B in CM derived from H2122 cells cocultured with H2122-specific CD8 + T cells ( n = 3). L, GSEA of T-cell migration involved in the immune response signature in H2122 cells transduced with DPP4 or LUC. P values were calculated using an unpaired two-tailed Student's t test. *, P < 0.05; ***, P < 0.001.
Article Snippet: To generate H2122-specific T cells, PBMCs were cocultured with mitomycin C (MMC)–treated H2122 cells in AIM-V medium (Invitrogen) supplemented with 3% human male AB serum (Innovative Research), 10 IU/mL IL2, and 10 ng/mL IL15.
Techniques: Mutagenesis, Western Blot, Transduction, Activity Assay, Luciferase, Activation Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Cell Migration Assay, Migration, Software, Two Tailed Test
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