Figure 4 from Cytotoxic NK Cells Impede Response to Checkpoint Immunotherapy in Melanoma with an Immune-Excluded Phenotype

<p>Pharmacologic targeting of NK cells. <b>A,</b> Uniform Manifold Approximation and Projection (UMAP) of re-clustered NK cells from the <i>NRAS;Ink4a</i> and YUMM5.2 tumors (left), selected genes expression between NK cell clusters (middle), and proportions of NK cells from the <i>NRAS;Ink4a</i> and YUMM5.2 tumors compared between control- and αPD-1–treated condition (right) based on the scRNA-seq experiment. <b>B,</b> Dot plot of the expression of selected genes between <i>NRAS;Ink4a</i> and YUMM5.2 tumors in NK cells only based on the scRNA-seq experiment. <b>C,</b> Volcano plot representing differentially expressed proteins in NK cells from the <i>NRAS;Ink4a</i> tumor between control and αPD-1 treatment. Significantly expressed proteins were considered based on the adjusted <i>P</i> value > 10e–3 based on the CITE-seq experiment. <b>D,</b> Growth curves (left) and doubling time calculation (right) of <i>NRAS;Ink4a</i> tumors <i>in vivo</i> treated with αPD-1, JMS-17-2, and CD38 (<i>n</i> ≥ 4 per cohort and a Welch-corrected <i>t</i> test was performed on the doubling time of tumors upon exponential fitting; **, <i>P</i> < 0.01). <b>E,</b> Quantification of immune cells (CD45<sup>+</sup>), NK cells (CD45<sup>+</sup>NKP46<sup>+</sup>), CD8<sup>+</sup> T cells (CD45<sup>+</sup>CD3<i>ε</i><sup>+</sup>CD8<i>α</i><sup>+</sup>), and macrophages (CD45<sup>+</sup>F4/80<sup>+</sup>) in multiplexed immunofluorescence images (each dot corresponds to a single tumor; data analyzed with a Welch-corrected <i>t</i> test; *, <i>P</i> < 0.05; **, <i>P</i> < 0.01). Tumor rim corresponds to an area ≤0.2 mm<sup>2</sup> in proximity to the tumor border.</p>