Detailed bladder cancer immunoprofiling reveals new clues for immunotherapeutic strategies

Abstract

ObjectivesBladder cancer (BlCa) is the tenth most frequent malignancy worldwide and the costliest to treat and monitor. Muscle‐invasive BlCa (MIBC) has a dismal prognosis, entailing the need for alternative therapies for the standard radical cystectomy. Checkpoint blockade immunotherapy has been approved for high‐grade non‐muscle‐invasive BlCa (HG NMIBC) and metastatic disease, but its effectiveness in localised MIBC remains under scrutiny. Herein, we sought to characterise and compare the immune infiltrate of HG NMIBC and MIBC samples, including ICOS expression, a targetable immune checkpoint associated with regulatory T cell(Tregs)‐mediated immunosuppression.MethodsImmunohistochemistry for CD83, CD20, CD68, CD163, CD3, CD8, CD4, FoxP3/ICOS and PD‐L1 was performed in HG NMIBC and MIBC samples (n = 206), and positive staining was quantified in the peritumoral and/or intratumoral tissue compartments with QuPath imaging software.ResultsCD20+ B cells, CD68+ and CD163+ tumor‐associated macrophages were significantly increased in MIBCs and associated with poor prognosis. In turn, higher infiltration of T cells was associated with prolonged survival, with exception of the CD4+ helper subset. Intratumoral expression of CD3 and CD8 was independent prognostic factors for increased disease‐free survival (DFS) in multivariable analysis. Remarkably, Tregs (FoxP3+/FoxP3+ICOS+) were found differentially distributed between tissue compartments. PD‐L1 immunoexpression independently predicted a shorter DFS and associated with higher infiltration of FoxP3+ICOS+ Tregs.ConclusionsImmune infiltrates of HG NMIBC and MIBC display significant differences that may help selecting patients for immunotherapies. Considering ICOS immunoexpression results, it might constitute a relevant therapeutic target, eventually in combination with anti‐PD‐1/PD‐L1 therapies, for certain BlCa patient subsets.

Tumor microenvironment and epithelial-mesenchymal transition in bladder cancer: Cytokines in the game?

“…In BlCa tissues, CD4 + levels were significantly higher in pTa-pT1 patients, comparing with most aggressive tumors (Jóźwicki et al, 2016;Viveiros et al, 2022). However, stratifying the tumoral areas, it was observed that CD4 + cells were significantly enriched in high-grade NMIBCs in peritumoral area, while CD4 + levels were significantly abundant in MIBCs in intratumoral area (Viveiros et al, 2022).…”

“…Specifically, in Zhang S et al study, higher CD8 + was associated with better OS in non-organ confined disease, but with worse OS in organ-confined disease patients, suggesting that cytotoxic T cells might have antitumor activity in non-organ confined disease and a protumor activity in organ-confined disease Zhang et al (2017). Viveiros N et al observed that MIBC patients presented higher CD8 + expression, comparing with NMIBC high-grade, but, specifically, MIBC tumors with high intratumoral CD8 expression demonstrated higher DFS and OS Viveiros et al (2022). Additionally, it was shown that poor CD8 + T cell expression, along with type I IFN signature and IFN-inducible inhibitory factors, characterize a non-T cell inflamed bladder TME (Trujillo et al, 2018), usually correlated with poor prognosis and resistance to immunotherapies (Sweis et al, 2016).…”

“…Overall T cells (CD3 + ) were significant increase in MIBC tumors, comparing with high-grade NMIBCs, although no differences were found in bladder peritumoral areas (Viveiros et al, 2022). It was shown that CD3 + in tumor infiltrating lymphocytes (TILs) were related with poor outcome in BlCa patients (Russo et al, 2022).…”

“…Zhang Y et al showed, in silico, that activated memory CD4 + cells were significantly associated with better outcome, while resting memory CD4 + cells were associated with poor outcome in BlCa patients Zhang et al (2020). In BlCa tissues, CD4 + levels were significantly higher in pTa-pT1 patients, comparing with most aggressive tumors (Jóźwicki et al, 2016;Viveiros et al, 2022). However, stratifying the tumoral areas, it was observed that CD4 + cells were significantly enriched in high-grade NMIBCs in peritumoral area, while CD4 + levels were significantly abundant in MIBCs in intratumoral area (Viveiros et al, 2022).…”

“…In BlCa, higher amounts of CD68 + (pan-macrophage marker) cells, were associated with higher grade and advanced tumors ( Huang et al, 2020 ; Harras and Abo Safia, 2021 ). Specifically, TAMs (CD68 + ) number was significantly higher in MIBCs comparing with NMIBCs ( Hanada et al, 2000 ; Viveiros et al, 2022 ) and higher amounts of CD68 + cells were significantly associated with poorer disease specific survival (DSS) in bladder peritumoral regions and with worse overall survival (OS) and DSS in bladder intratumoral regions ( Viveiros et al, 2022 ). Co-cultures between macrophages and BlCa cell lines showed an increase in colony formation, cell migration and cell invasion ( Huang et al, 2020 ).…”