Prognostic significance of myeloid immune cells and their spatial distribution in the colorectal cancer microenvironment

Abstract

<div lang="en" class="abs"><p class="abs_header">Abstract</p><p><em>Background:</em> Myeloid cells represent an abundant yet heterogeneous cell population in the colorectal cancer microenvironment, and their roles remain poorly understood.</p><p><em>Methods:</em> We used multiplexed immunofluorescence combined with digital image analysis to identify CD14⁺ monocytic and CD15⁺ granulocytic cells and to evaluate their maturity (HLA-DR and CD33), immunosuppressive potential (ARG1) and proximity to cytokeratin (KRT)-positive tumor cells in 913 colorectal carcinomas. Using covariate data of 4465 incident colorectal cancers in two prospective cohort studies, the inverse probability weighting method was used with multivariable-adjusted Cox proportional hazards models to assess cancer-specific mortality according to ordinal quartiles (Q1–Q4) of myeloid cell densities. Immune cell–tumor cell proximity was measured with the nearest neighbor method and the G-cross function, which determines the likelihood of any tumor cell having at least one immune cell of the specified type within a certain radius.</p><p><em>Results:</em> Higher intraepithelial (<em>P_trend</em>=0.0002; HR for Q4 (vs Q1), 0.48, 95% CI 0.31 to 0.76) and stromal (Ptrend <0.0001; HR for Q4 (vs Q1), 0.42, 95% CI 0.29 to 0.63) densities of CD14⁺HLA-DR⁺ cells were associated with lower colorectal cancer-specific mortality while, conversely, higher intraepithelial densities of CD14⁺HLA-DR⁻ cells were associated with higher colorectal cancer-specific mortality (<em>P_trend</em>=0.0003; HR for Q4 (vs Q1), 1.78, 95% CI 1.25 to 2.55). Spatial analyses indicated that CD15⁺ cells were located closer to tumor cells than CD14⁺ cells, and CD14⁺HLA-DR⁺ cells were closer to tumor than CD14⁺HLA-DR⁻ cells (p<0.0001). The G-cross proximity measurement, evaluating the difference in the likelihood of any tumor cell being colocated with at least one CD14⁺HLA-DR⁺ cell versus CD14⁺HLA-DR⁻ cell within a 20 µm radius, was associated with lower colorectal cancer-specific mortality (<em>P_trend</em> <0.0001; HR for Q4 (vs Q1), 0.37, 95% CI 0.24 to 0.57).</p><p><em>Conclusions:</em> Myeloid cell populations occur in spatially distinct distributions and exhibit divergent, subset-specific prognostic significance in colorectal cancer, with mature CD14⁺HLA-DR⁺ and immature CD14⁺HLA-DR⁻ monocytic phenotypes most notably showing opposite associations. These results highlight the prognostic utility of multimarker evaluation of myeloid cell infiltrates and reveal a previously unrecognized degree of spatial organization for myeloid cells in the immune microenvironment.</p></div>