Time-sequential change in immune-related gene expression after irradiation in glioblastoma: next-generation sequencing analysis

Anim Cells Syst (Seoul). 2021 Jul 30;25(4):245-254. doi: 10.1080/19768354.2021.1954550. eCollection 2021.

Abstract

The time-sequential change in immune-related gene expression of the glioblastoma cell line after irradiation was evaluated to speculate the effect of combined immunotherapy with radiotherapy. The U373 MG glioblastoma cell line was irradiated with 6 Gy single dose. Next-generation sequencing (NGS) transcriptome data was generated before irradiation (control), and at 6, 24, and 48 h post-irradiation. Immune-related pathways were analyzed at each time period. The same analyses were also performed for A549 lung cancer and U87 MG glioblastoma cell lines. Western blotting confirmed the programmed death-ligand 1 (PD-L1) expression levels over time. In the U373 MG cell line, neutrophil-mediated immunity, type I interferon signaling, antigen cross-presentation to T cell, and interferon-γ signals began to increase significantly at 24 h and were upregulated until 48 h after irradiation. The results were similar to those of the A549 and U87 MG cell lines. Without T cell infiltration, PD-L1 did not increase even with upregulated interferon-γ signaling in cancer cells. In conclusions, in the glioblastoma cell line, immune-related signals were significantly upregulated at 24 and 48 h after irradiation. Therefore, the time interval between daily radiotherapy might not be enough to expect full immune responses by combined immune checkpoint inhibitors and newly infiltrating immune cells after irradiation.

Keywords: Glioblastoma; Immune-related signals; Next-generation sequencing; Radiotherapy; Transcriptome.

Grants and funding

This work was supported by the National Research Foundation of Korea (NRF, https://www.nrf.re.kr/eng/index) grant funded by the Korea government (MSIP) [grant number 2013M2A2A7043683] to IHK, the Ministry of Science and ICT [grant number 2020R1C1C1011369] to YJK, and the Ewha Alumni Medical Research [grant number 2018] to YJK.