Fig. 2

The expression of genes required for PD-1 signaling is associated with clinical response to PD-1 blockade. Diagram of the experimental pipeline from transduction of the Jurkat T cells, cell labeling, stimulation, sorting, and guide sequencing (A). Venn diagram showing the number of genes identified by sequencing in the two groups of sorted cells (B). Graph showing the levels of secreted IL-2 measured in each clone of Jurkat short hairpin knockdown T cells under the indicated treatment conditions, n = 4 for each cell line; average values are shown (C). Heatmaps showing the differential expression levels of genes that we discovered in our screen to be involved in PD-1 downstream signaling, now in T cells isolated from cancer patients that were treated with anti-PD-1 antibodies in three separated clinical trials (SKCM1, BCC, SCC) (D). The patients were divided into those who responded favorably to PD-1 blockade (R) and those who failed to respond (NR). Volcano plots display expression levels of genes that were either upregulated or downregulated in the cancer patients who failed to respond to PD-1 blockade (E). Differences in the expression levels were calculated by implementing Limma statistics. GO-enrichment analysis of the genes identified in the screen and involved in the PD-1 signaling pathway (F). A berswarm plot expression levels of genes discovered in the screen and as differentially expressed in the T cell from the cancer patients subjected to PD-1 blockade (G). Bolded genes are those that were shared in all three clinical trials