Bulk RNA-Seq analysis of adoptively co-transferred PKM2 wildtype and PKM2 knockout CD8+ T cells into tumor-bearing mice subsequently treated with and without PD-1 checkpoint blockade

In this study we investigated how manipulation of the glycolytic enzyme pyruvate kinase muscle 2 (PKM2) alters CD8+ T cell gene expression and responsiveness to PD-1 checkpoint blockade in orthotopic mouse lung adenocarcinoma. In this dataset, we performed adoptive co-transfers of antigen-specific OT-I+ Thy1.1+ CD8+ T cells genetically modified by CRISPR/Cas9 ribonucleoproteins to become PKM2 knockout (in Thy1.1+/- cells) or PKM2 wildtype (in Thy1.1+/+ cells). T cells were transferred into orthotopic mouse lung adenocarcinoma HKP1-ova-GFP tumor-bearing syngeneic C57BL/6 mice, and subsequently mice were treated with either IgG or anti-PD-1. Adoptively-transferred T cells were sorted based on Thy1.1 zygosity from tumors at two different time points and bulk RNA sequenced. Comparative gene expression profiling of three biological replicates of adoptively co-transferred PKM2 wild-type or PKM2 knockout OT-I+ CD8+ T cells, distinguished by Thy1.1 zygosity, into Thy1.2+/+ C57BL/6 mice bearing HKP1-ova-GFP orthotopic lung tumors and subsequently treated with either IgG or anti-PD-1, harvested at two timepoints.