Together Intra-Tumor Hypoxia and Macrophagic Immunity Are Driven Worst Outcome in Pediatric High-Grade Osteosarcomas

SIMPLE SUMMARY: Radiological and immunohistochemical data were correlated with the outcome in a retrospective monocentric cohort of 30 pediatric osteosarcomas (OTS). A necrotic volume of more than 50 cm(3) at diagnosis was significantly linked to a worse overall survival (OS). Regarding immunohistochemical analyses, an overexpression of hypoxic markers, such as HIF-1α and anhydrase carbonic IX (CAIX), was significantly linked to a worse OS, while pS6-RP hyperexpression was correlated with a better survival. We also featured that CD68 positive cells, representative of macrophagic M1 polarization, were mostly associated with HIF-1α and CAIX hyperexpressions and that M2-like polarization, mostly related to CD163 positivity, was correlated to mTor activation. These findings, involving clinical, radiological and biology data, allowed us to hypothesize a dual signature association ready to use routinely in future protocols. ABSTRACT: Background: Osteosarcomas (OTS) represent the most common primary bone cancer diagnosed in adolescents and young adults. Despite remarkable advances, there are no objective molecular or imaging markers able to predict an OTS outcome at diagnosis. Focusing on biomarkers contributing broadly to treatment resistance, we examine the interplay between the tumor-associated macrophages and intra-tumor hypoxia. Methods: Radiological and immunohistochemical (IHC) data were correlated with the outcome in a retrospective and monocentric cohort of 30 pediatric OTS. We studied hypoxic (pS6, phospho-mTor, HIF-1α and carbonic anhydrase IX (CAIX)) and macrophagic (CD68 and CD163) biomarkers. Results: The imaging analyses were based on MRI manual volumetric measures on axial post-contrast T1 weighted images, where, for each tumor, we determined the necrotic volume and its ratio to the entire tumor volume. When they were above 50 cm(3) and 20%, respectively, they correlated with a worse overall survival (p = 0.0072 and p = 0.0136, respectively) and event-free survival (p = 0.0059 and p = 0.0143, respectively). IHC assessments enable a significant statistical link between HIF-1α/CAIX hyper-expressions, CD68+ cells and a worse outcome, whereas activation of mTor pathway was linked to a better survival rate and CD163+ cells. Conclusions: This study evidenced the links between hypoxia and immunity in OTS, as their poor outcome may be related to a larger necrotic volume on diagnostic MRI and, in biopsies, to a specific IHC profile.