Fig. 4: Mitigating injury signalling inside peripheral sensory (nociceptors) neurons ameliorated intratumoural immunosuppression.
From: Cancer-induced nerve injury promotes resistance to anti-PD-1 therapy
a, Uniform manifold approximation and projection (UMAP) plot of scRNA-seq data of DRG neurons innervating mouse paws that were inoculated with either B16F10-OVA melanoma or normal keratinocytes (Extended Data Fig. 9a). Clustering analysis revealed a subgroup of neurons termed cancer-injured neurons (CIN), representing 4.5% (710 neurons) of the melanoma group (total, 15,818 neurons) versus less than 0.1% (12 neurons) of the keratinocyte group (total, 17,133 neurons). KIN, keratinocyte-innervating neurons; MIN, melanoma-innervating neurons; SST neurons, somatostatin-expressing neurons; uc, unnamed cluster. b, Cancer-injured neurons were characterized by an enrichment of nerve-injury-related genes. c, Tumour growth curve comparing the tumour growth of melanomas inoculated into mice with a cre-flox conditional knockout of Atf3 in nociceptor neurons (Atf3-cKO, n = 7) versus their permissive littermate controls (NaV1.8WT::Atf3fl/fl, WT, n = 6). Data are mean ± s.e.m. Tumour growth over time (P < 0.001) was analysed using a mixed-effects model; post hoc comparisons for individual timepoints were conducted using Šídák’s multiple-comparison test (P < 0.001). d, Flow-cytometry-based assessment of IFNγ+CD8+ T cells in WT (n = 5) versus Atf3-cKO (n = 4) mice. The box plots show the median value (centre line), the 25th and 75th percentiles (box limits), and the minimum and maximum values (whiskers). Statistical analysis was performed using non-paired two-tailed Student’s t-tests. e, scRNA-seq analysis of intratumoural (melanoma) immune (CD45+) cells in Atf3-cKO versus WT mice (Extended Data Fig. 10) showing major immune cell compositions. f, Expression of Lag3 and Tox genes in CD8+ T cells from the immune scRNA-seq experiment presented in e. g, Expression of M1 macrophage markers (Il1b+Cd86+Cd80+, associated with enhanced antitumoural immune activity) in intratumoural macrophages. Statistical analysis was performed using two-tailed Wilcoxon rank-sum tests; significance was established through normal approximation. For Il1b, P = 0.006; for Cd86, P = 0.022; and for Cd80, P = 0.0003. h, Significantly enriched gene sets, based on the WikiPathways dataset among intratumoural macrophages. ORA, over-representation analysis.
