Figure 6.

CyC directs recruitment of TREM2+ macrophages and promotes failure of cancer immunotherapy

(A) Tumor growth curves (mean and standard error of the mean) for single-flank sgScrambled (n = 8) and CST3^−/− (CST3 knockout [KO], n = 8) tumors; 100,000 cells were inoculated in right flank (cohort A).

(B) Tumor growth curves (mean and standard error of the mean) for biflank paired sgScrambled (n = 5) and CST3^−/− (n = 5) tumors; 50,000 cells were inoculated in both flanks (cohort C). Mice received three doses of anti-PD-L1 antibody. p values refer to paired two-sided t tests.

(C and D) Proliferation index (proportion of Ki67+ cells/total cells) (C) and proportion (D) of non-epithelial cells in histological sections from paired biflank sgScrambled and CST3^−/− tumors (pooled cohorts C and D). p values refer to paired two-sided t tests.

(E) Uniform manifold approximation and projection (UMAP) of 14,416 cells, annotated with cell type, from 4 tumor samples (2 sgScrambled, 2 CST3^−/−).

(F) Proportion of Trem2+ macrophages in sgScrambled and CST3^−/− tumors; p value is adjusted p value from linear model of logit-transformed proportions.

(G) Number of Trem2+ cells per mm² from digital image analysis of Trem2 immunohistochemistry in paired biflank sections from sgScrambled and CST3^−/− tumors. p value refers to paired two-sided t test.

(H) Multivariate (Cox and logistic) regression of Z scored CyC-production PGS against immuno-oncology biomarkers (progression-free survival [PFS], overall survival [OS], durable clinical benefit [DCB]) in meta-analysis of European patients (n = 685) treated with checkpoint immunotherapy (anti-CTLA4 or anti-PD1/PD-L1). Sample sizes for each clinical endpoint were n = 342, 685, and 670, respectively. In each model, covariates included PC1–4, sex, and primary cancer. Error bars reflect 95% confidence interval. Lower hazard ratios (survival, Cox regression) or higher odds ratios (durable clinical benefit, logistic regression) reflect better therapeutic outcomes (annotated with purple arrow).

(I) Sensitivity analysis indicating odds ratio and 95% confidence interval for DCB in each cancer type. p values refer to two-sided t tests unless otherwise stated. ∗p < 0.05; ∗∗p < 0.01, ∗∗∗p < 0.001.

Boxplots show median (central line) with interquartile range (IQR; box) and extrema (whiskers at 1.5× IQR).