Basic Study
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World J Gastroenterol. Jun 7, 2022; 28(21): 2302-2319
Published online Jun 7, 2022. doi: 10.3748/wjg.v28.i21.2302
Impact of radiotherapy on the immune landscape in oesophageal adenocarcinoma
Noel E Donlon, Maria Davern, Fiona O’Connell, Andrew Sheppard, Aisling Heeran, Anshul Bhardwaj, Christine Butler, Ravi Narayanasamy, Claire Donohoe, James J Phelan, Niamh Lynam-Lennon, Margaret R Dunne, Stephen Maher, Jacintha O’Sullivan, John V Reynolds, Joanne Lysaght
Noel E Donlon, Maria Davern, Fiona O’Connell, Andrew Sheppard, Aisling Heeran, Anshul Bhardwaj, Christine Butler, Ravi Narayanasamy, Claire Donohoe, James J Phelan, Niamh Lynam-Lennon, Margaret R Dunne, Stephen Maher, Jacintha O’Sullivan, John V Reynolds, Joanne Lysaght, Department of Surgery, Trinity Translational Medicine Institute, St James Hospital, Dublin D08, Ireland
Author contributions: Donlon NE and Davern M contributed equally to this work; Donlon NE and Davern M contributed to experimental design and execution, and manuscript drafting and revision; O’Connell F and Sheppard A contributed to experiments; Heeran A, Bhardwaj A, and Butler C contributed to sample acquisition; Narayanasamy R, Donohoe C, Phelan JJ, Lynam-Lennon N, Dunne MR, and Maher S contributed to concept design; Phelan JJ contributed to statistical analysis; O’Sullivan J, Reynolds JV, and Lysaght J contributed to paper revision and supervision of the project.
Institutional review board statement: The study was reviewed and approved by the Tallaght/St James’s Hospital Ethics Committee.
Conflict-of-interest statement: There are no conflicts of interest to report.
Data sharing statement: Data generated in this study will be available upon specific request from the corresponding author.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Noel E Donlon, MD, Assistant Professor, Department of Surgery, Trinity Translational Medicine Institute, St James Hospital, James’s Street, Dublin D08, Ireland. donlonn@tcd.ie
Received: December 3, 2021
Peer-review started: December 3, 2021
First decision: January 27, 2022
Revised: February 19, 2022
Accepted: April 26, 2022
Article in press: April 26, 2022
Published online: June 7, 2022
Processing time: 180 Days and 14.7 Hours
Abstract
BACKGROUND

In the contemporary era of cancer immunotherapy, an abundance of clinical and translational studies have reported radiotherapy (RT) and immunotherapies as a viable option for immunomodulation of many cancer subtypes, with many related clinical trials ongoing. In locally advanced disease, chemotherapy or chemoradiotherapy followed by surgical excision of the tumour remain the principal treatment strategy in oesophageal adenocarcinoma (OAC), however, the use of the host immune system to improve anti-tumour immunity is rapidly garnering increased support in the curative setting.

AIM

To immunophenotype OAC patients’ immune checkpoint (IC) expression with and without radiation and evaluate the effects of checkpoint blockade on cell viability.

METHODS

In the contemporary era of cancer immunotherapy, an abundance of studies have demonstrated that combination RT and IC inhibitors (ICIs) are effective in the immunomodulation of many cancer subtypes, with many related clinical trials ongoing. Although surgical excision and elimination of tumour cells by chemotherapy or chemoradiotherapy remains the gold standard approach in OAC, the propagation of anti-tumour immune responses is rapidly garnering increased support in the curative setting. The aim of this body of work was to immunophenotype OAC patients’ IC expression with and without radiation and to establish the impact of checkpoint blockade on cell viability. This study was a hybrid combination of in vitro and ex vivo models. Quantification of serum immune proteins was performed by enzyme-linked immunosorbent assay. Flow cytometry staining was performed to evaluate IC expression for in vitro OAC cell lines and ex vivo OAC biopsies. Cell viability in the presence of radiation with and without IC blockade was assessed by a cell counting kit-8 assay.

RESULTS

We identified that conventional dosing and hypofractionated approaches resulted in increased IC expression (PD-1, PD-L1, TIM3, TIGIT) in vitro and ex vivo in OAC. There were two distinct subcohorts with one demonstrating significant upregulation of ICs and the contrary in the other cohort. Increasing IC expression post RT was associated with a more aggressive tumour phenotype and adverse features of tumour biology. The use of anti-PD-1 and anti-PD-L1 immunotherapies in combination with radiation resulted in a significant and synergistic reduction in viability of both radiosensitive and radioresistant OAC cells in vitro. Interleukin-21 (IL-21) and IL-31 significantly increased, with a concomitant reduction in IL-23 as a consequence of 4 Gray radiation. Similarly, radiation induced an anti-angiogenic tumour milieu with reduced expression of vascular endothelial growth factor-A, basic fibroblast growth factor, Flt-1 and placental growth factor.

CONCLUSION

The findings of the current study demonstrate synergistic potential for the use of ICIs and ionising radiation to potentiate established anti-tumour responses in the neoadjuvant setting and is of particular interest in those with advanced disease, adverse features of tumour biology and poor treatment responses to conventional therapies.

Keywords: Oesophageal Cancer; Radiotherapy; Immunotherapy; Immunology; Surgery; Oncology

Core Tip: This body of work evaluates the impact of radiotherapy on the immune profile in oesophageal adenocarcinoma with an added caveat of immunotherapy effects on tumour cell killing.