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Related Experiment Video

Updated: Nov 5, 2025

Visualizing and Quantifying Endonuclease-Based Site-Specific DNA Damage

Visualizing and Quantifying Endonuclease-Based Site-Specific DNA Damage

Published on: August 21, 2021

ROS induces NETosis by oxidizing DNA and initiating DNA repair.

Dhia Azzouz^1,2, Meraj A Khan¹, Nades Palaniyar^3,4,5

¹Program in Translational Medicine, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children, Toronto, ON, Canada.

Cell Death Discovery

|May 18, 2021

Summary

This summary is machine-generated.

Reactive oxygen species (ROS) drive neutrophil extracellular trap formation (NETosis) by causing DNA damage. DNA repair initiation, not completion, triggers NETosis via chromatin decondensation.

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Area of Science:

Immunology
Molecular Biology
Cell Biology

Background:

Reactive oxygen species (ROS) are crucial for neutrophil extracellular trap (NET) formation, a process known as NETosis.
The precise mechanism by which ROS induces NETosis remains largely unknown.
Neutrophil activation generates substantial ROS and non-coding transcription, facilitating chromatin decondensation.

Purpose of the Study:

To elucidate the mechanism linking ROS production to NETosis induction.
To investigate the role of DNA damage and repair in ROS-mediated NETosis.

Main Methods:

Induction of NADPH oxidase-dependent NETosis using various agonists (PMA, E. coli LPS, S. aureus, P. aeruginosa).
Assessment of DNA damage and translocation of proliferating cell nuclear antigen (PCNA).
Inhibition of DNA repair pathways (APE1, PARP, DNA ligase) and evaluation of NETosis suppression.

Main Results:

NADPH oxidase-dependent NETosis induction results in extensive DNA damage, including guanine oxidation to 8-oxoguanine.
DNA repair protein PCNA translocates from the cytoplasm to the nucleus during NETosis.
Inhibiting early DNA repair steps (e.g., APE1, PARP, DNA ligase) suppresses NETosis.
Later repair steps involving DNA polymerases and PCNA do not inhibit agonist-induced NETosis.

Conclusions:

Excess ROS during neutrophil activation induces NETosis by causing significant DNA damage.
The subsequent initiation of the DNA repair pathway, leading to chromatin decondensation, is the critical step in ROS-induced NETosis.
Targeting early DNA repair mechanisms may offer a strategy to control NETosis.