Short-course radiotherapy promotes pro-inflammatory macrophages via extracellular vesicles in human rectal cancer

Abstract

BackgroundTumor-associated macrophages (TAM) constitute the most abundant immune cells in the tumor stroma initiating pro-inflammatory (M1) or immunosuppressive (M2) responses depending on their polarization status. Advances in tumor immunotherapy call for a detailed understanding of potential immunogenic mechanisms of irradiation routinely applied in rectal cancer patients.MethodsTo test the effects of radiotherapy on TAM, we ex vivo irradiated tissue samples of human rectal cancer and assessed the phenotype by flow cytometry. We furthermore evaluated the distribution of leucocyte subsets in tissue sections of patients after short-course radiotherapy and compared findings to non-pretreated rectal cancer using an immunostaining approach. Organotypic assays (OTA) consisting of macrophages, cancer-associated fibroblast and cancer cell lines were used to dissect the immunological consequences of irradiation in macrophages.ResultsWe demonstrate that short-course neoadjuvant radiotherapy in rectal cancer patients is associated with a shift in the polarization of TAM towards an M1-like pro-inflammatory phenotype. In addition, ex vivo irradiation caused an increase in the phagocytic activity and enhanced expression of markers associated with stimulatory signals necessary for T-cell activation. In OTA we observed that this alteration in macrophage polarization could be mediated by extracellular vesicles (EV) derived from irradiated tumor cells. We identified high mobility group box 1 in EV from irradiated tumor cells as a potential effector signal in that crosstalk.ConclusionsOur findings highlight macrophages as potential effector cells upon irradiation in rectal cancer by diminishing their immunosuppressive phenotype and activate pro-inflammation. Our data indicate that clinically applied short-term radiotherapy for rectal cancer may be exploited to stimulate immunogenic macrophages and suggest to target the polarization status of macrophages to enhance future immunotherapeutic strategies.

Systematic review of the immunological landscape of Wilms tumors

“… 92 Radiotherapy was shown to induce a re-programming of M2-like macrophages into M1-like macrophages. 93 , 94 The latter aspect could be specifically relevant in WT, as M2 macrophages are predominant. As neo-adjuvant radio- and chemotherapy are already included in the standard treatment for many pediatric solid cancers, it would be crucial to understand their putative immunomodulatory effects in this tumor entity and eventually consider them for a coherent combination therapy with checkpoint inhibitors.…”

Emerging Trends for Radio-Immunotherapy in Rectal Cancer

“…SCPRT (2 × 5 Gy) was also found to be associated with a shift in the polarization of TAMs towards an M1-like pro-inflammatory phenotype in vitro and in vivo [ 165 ]. Similarly, in another study, irradiated macrophages (5 × 2 Gy) showed a reduced anti-inflammatory profile, increased phagocytosis and unaltered pro-invasive and pro-angiogenic capacities [ 166 ].…”

Radiation Response in the Tumour Microenvironment: Predictive Biomarkers and Future Perspectives

“…Clinically, in patients with T3 rectal cancer, a short course of radiotherapy (neoadjuvant hyperfractionated 25 Gy in 10 fractions; surgery performed on day 2–5) promoted TAM repolarisation towards an M1-like proinflammatory phenotype. Interestingly, ex vivo modelling of this response suggested that HMGB1 in EVs from irradiated tumour cells could be responsible for this effect on TAM polarisation [ 38 ].…”