Corticosteroid-dependent association between prognostic peripheral blood cell-free DNA levels and neutrophil-mediated NETosis in patients with glioblastoma.

in Clinical cancer research : an official journal of the American Association for Cancer Research by Jacob E Till, Nicholas J Seewald, Zachariya Yazdani, Zhuoyang Wang, Dominique Ballinger, Heather Samberg, Siri Dandu, Camilla Macia, Melinda Yin, Aseel Abdalla, Timothy Prior, Shivani Shah, Thara Patel, Emily McCoy, Maikel Mansour, Carson A Wills, Veronica Bochenek, Jonathan Serrano, Matija Snuderl, Richard E Phillips, Donald M O'Rourke, Nduka M Amankulor, Ali Nabavizadeh, Arati S Desai, Kandace Gollomp, Zev A Binder, Wanding Zhou, Stephen J Bagley, Erica L Carpenter

TLDR

  • The study found that neutrophil-mediated NETosis is the dominant source of prognostic ccfDNA in GBM patients and may be associated with glucocorticoid exposure.

Abstract

Non-invasive prognostic biomarkers to inform clinical decision-making are an urgent unmet need for the management of patients with glioblastoma (GBM). We previously showed that higher circulating cell-free DNA concentration [ccfDNA] is associated with worse survival in GBM. However, the biology underlying this is unknown. We prospectively enrolled 129 patients with treatment-naïve GBM with blood drawn prior to initial resection (baseline) and at time of first post-radiotherapy MRI. We performed ccfDNA methylation deconvolution to determine cellular sources of ccfDNA. ELISA was performed to detect citrullinated H3 (citH3), a marker of neutrophil extracellular traps (NETs). Multiplex proteomic analysis was used to measure soluble inflammatory proteins. We found that neutrophils contributed the highest proportion of prognostic ccfDNA. The percentage of ccfDNA derived from neutrophils was correlated with total [ccfDNA], but only in patients receiving pre-operative corticosteroids. At baseline and on-therapy, [citH3] was significantly higher in the plasma of patients with GBM receiving corticosteroids compared to corticosteroid-naïve GBMs or no-cancer controls. Unsupervised hierarchical clustering of ccfDNA methylation patterns yielded two clusters, with one enriched for patients with the NETosis phenotype and who received corticosteroids. Unsupervised clustering of circulating inflammatory proteins yielded similar results. These data suggest neutrophil-mediated NETosis is the dominant source of prognostic ccfDNA in patients with GBM and may be associated with glucocorticoid exposure. If further studies show that pharmacological inhibition of NETosis can mitigate the deleterious effects of corticosteroids, these plasma markers will have important clinical utility as non-invasive correlative biomarkers.

Overview

  • The study investigated the source and significance of circulatory cell-free DNA (ccfDNA) in glioblastoma (GBM) patients.
  • The study used ccfDNA methylation deconvolution to determine the cellular sources of ccfDNA and found that neutrophils contributed the highest proportion of prognostic ccfDNA.
  • The study aimed to determine the significance of neutrophil-mediated NETosis as a potential biomarker for GBM diagnosis and treatment.

Comparative Analysis & Findings

  • The study found that the percentage of ccfDNA derived from neutrophils was correlated with total ccfDNA only in patients receiving pre-operative corticosteroids.
  • At baseline and on-therapy, citrullinated H3 (citH3) was significantly higher in the plasma of patients with GBM receiving corticosteroids compared to corticosteroid-naïve GBMs or no-cancer controls.
  • Unsupervised hierarchical clustering of ccfDNA methylation patterns yielded two clusters, with one enriched for patients with the NETosis phenotype and who received corticosteroids.

Implications and Future Directions

  • The study suggests that pharmacological inhibition of NETosis may be a potential therapeutic strategy to mitigate the deleterious effects of corticosteroids in GBM patients.
  • Further studies are needed to determine the role of NETosis in the development and progression of GBM and to investigate the potential therapeutic applications of NETosis inhibition.
  • The study highlights the potential of ccfDNA-based biomarkers, including neutrophil-mediated NETosis, for non-invasive management of GBM patients.
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