Fig. 8 |. A low myeloid response is associated with worse patient outcome in ACC.

UMAP of scRNA-seq data from adrenal tissue of female Znrf3-cKO animals. Expression of individual myeloid cell markers Cd33 (a), Cd68 (b), Itgam (also known as CD11b) (c) and Msr1 (also known as CD204) (d) is shown. e, The combined expression of these four markers was used to generate an AMRS. Exp, expression. f, Analysis of TCGA-ACC data, including patient demographics and outcome, in accordance with mRNA expression of AMRS genes (CD33, CD68, ITGAM, MSR1) and established prognostic markers (high MKI67 (proliferation) and low G0S2 (recurrent disease)). AMRS is significantly lower in female than in male TCGA patients with ACC (g), and low AMRS is associated with shorter overall (h) and progression-free (i) survival. j, In primary human ACC tumors, CD68⁺ histiocytes are found in a subset of cases. k, A representative example region is shown relative to a Znrf3-cKO mouse adrenal tumor. l,m, The infiltration of CD68⁺ myeloid cells in primary human ACC tumors is heterogeneous. Regions with a high Ki67 index (>15%) have a significantly lower CD68 index than regions of low proliferation (Ki67 < 5%). IHC quantification was performed using QuPath digital analysis based on the number of positive cells normalized to total nuclei. Tumor-associated stroma and highly necrotic regions were excluded from analysis. Each dot represents a distinct tumor region (n = 50 tumor regions from 38 patients). Statistical analysis was performed using two-tailed Mann–Whitney test (g), log-rank Mantel–Cox test (h,i) or two-tailed Student’s t-test (m). Scale bars, 100 μm.