Spatial Clustering of CD68+ Tumor Associated Macrophages with Tumor Cells is Associated with Worse Overall Survival in Metastatic Clear Cell Renal Cell Carcinoma

Abstract

Immune infiltration is typically quantified using cellular density, not accounting for cellular clustering. Tumor-associated macrophages (TAM) activate oncogenic signaling through paracrine interactions with tumor cells, which may be better reflected by local cellular clustering than global density metrics. Using multiplex immunohistochemistry and digital pathologic analysis we quantified cellular density and cellular clustering for myeloid cell markers in 129 regions of interest from 55 samples from 35 patients with metastatic ccRCC. CD68+ cells were found to be clustered with tumor cells and dispersed from stromal cells, while CD163+ and CD206+ cells were found to be clustered with stromal cells and dispersed from tumor cells. CD68+ density was not associated with OS, while high tumor/CD68+ cell clustering was associated with significantly worse OS. These novel findings would not have been identified if immune infiltrate was assessed using cellular density alone, highlighting the importance of including spatial analysis in studies of immune cell infiltration of tumors.SIGNIFICANCEIncreased clustering of CD68+ TAMs and tumor cells was associated with worse overall survival for patients with metastatic ccRCC. This effect would not have been identified if immune infiltrate was assessed using cell density alone, highlighting the importance of including spatial analysis in studies of immune cell infiltration of tumors.

Geospatial characterization of immune cell distributions and dynamics across the microenvironment in renal cell carcinoma

“…Most hypothesized mechanisms for the M2-like TAM pro-tumor effect rely on close proximity to their effector cell, and as such we aimed to associate measures of geospatial TAM clustering with clinical outcomes. Previous work by our group identified worse OS in patients with increased interface clustering of tumor cells and CD68+ TAMs, a pan-macrophage marker, in a small cohort of patients with metastatic ccRCC who subsequently received IT, further supporting the notion that TAM spatial distribution can affect outcomes 25 . In the present study, we identified more advanced clinical stage and substantially worse OS and CSS for patients with CD163+ M2-like TAMs clustered into the stromal compartment at the tumor-stroma interface.…”

“…However, scRNA-seq methodologies are tissue-destructive, impeding the ability to analyze the effect of immune cell geospatial location and distribution. TAMs affect the TIME through a variety of receptor-binding and paracrine mechanisms that rely on proximity to their effector cells, and the specific location and geospatial distribution of TAMs has been shown to have a significant impact on their pro-tumor activity [24][25][26][27] .…”

A Cuproptosis-Related lncRNAs Signature Could Accurately Predict Prognosis in Patients with Clear Cell Renal Cell Carcinoma

“…Besides, this group demonstrated greater expressions of inflammation-promoting and parainflammation factors as well as more intense type I Interferon (IFN) response, indicating a decreased autoimmunity function and a poor prognosis. If tumor-associated macrophages reveal a high infiltration, a poor OS in metastatic ccRCC could be expected [39]. In ccRCC patients, tumorinfiltrating mast cells secret IL-10 and TGF-β to decrease anti-tumor immunity, leading to a worse prognosis [40].…”

Three-dimensional tumour microenvironment reconstruction and tumour-immune interactions' analysis

“…More specifically, we used the localized segmented cell centres and a radius ] 25μ in order to quantify the number of immune cells. We chose this radius as a biologically relevant distance for interaction between tumour cells and immune cells [27]. Thus, we counted the number of the different immune cells that were identified within this radius of each segmented tumour cell.…”