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Journal of Experimental & Clinical Cancer Research

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01-12-2021 | Radiotherapy | Research

Interferon- and STING-independent induction of type I interferon stimulated genes during fractionated irradiation

Authors: Ruben S. A. Goedegebuure, Esther A. Kleibeuker, Francesca M. Buffa, Kitty C. M. Castricum, Syed Haider, Iris A. Schulkens, Luuk ten Kroode, Jaap van den Berg, Maarten A. J. M. Jacobs, Anne-Marie van Berkel, Nicole C. T. van Grieken, Sarah Derks, Ben J. Slotman, Henk M. W. Verheul, Adrian L. Harris, Victor L. Thijssen

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2021

Abstract

Background

Improvement of radiotherapy efficacy requires better insight in the dynamic responses that occur during irradiation. Here, we aimed to identify the molecular responses that are triggered during clinically applied fractionated irradiation.

Methods

Gene expression analysis was performed by RNAseq or microarray analysis of cancer cells or xenograft tumors, respectively, subjected to 3–5 weeks of 5 × 2 Gy/week. Validation of altered gene expression was performed by qPCR and/or ELISA in multiple cancer cell lines as well as in pre- and on-treatment biopsies from esophageal cancer patients (NCT02072720). Targeted protein inhibition and CRISPR/Cas-induced gene knockout was used to analyze the role of type I interferons and cGAS/STING signaling pathway in the molecular and cellular response to fractionated irradiation.

Results

Gene expression analysis identified type I interferon signaling as the most significantly enriched biological process induced during fractionated irradiation. The commonality of this response was confirmed in all irradiated cell lines, the xenograft tumors and in biopsies from esophageal cancer patients. Time-course analyses demonstrated a peak in interferon-stimulated gene (ISG) expression within 2–3 weeks of treatment. The response was accompanied by a variable induction of predominantly interferon-beta and/or -lambda, but blocking these interferons did not affect ISG expression induction. The same was true for targeted inhibition of the upstream regulatory STING protein while knockout of STING expression only delayed the ISG expression induction.

Conclusions

Collectively, the presented data show that clinically applied fractionated low-dose irradiation can induce a delayed type I interferon response that occurs independently of interferon expression or STING signaling. These findings have implications for current efforts that aim to target the type I interferon response for cancer treatment.

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Title: Interferon- and STING-independent induction of type I interferon stimulated genes during fractionated irradiation
Authors: Ruben S. A. Goedegebuure
Esther A. Kleibeuker
Francesca M. Buffa
Kitty C. M. Castricum
Syed Haider
Iris A. Schulkens
Luuk ten Kroode
Jaap van den Berg
Maarten A. J. M. Jacobs
Anne-Marie van Berkel
Nicole C. T. van Grieken
Sarah Derks
Ben J. Slotman
Henk M. W. Verheul
Adrian L. Harris
Victor L. Thijssen
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Radiotherapy
Interferon
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2021
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/s13046-021-01962-2

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