Introduction:

 NETs (Neutrophil Extracellular Traps) are web-like DNA structures formed by

nuclear proteins like actin, granule proteins, and histones.
 NETosis is a planned cell death pathway distinct from necrosis and apoptosis,
triggered by the discharge of extracellular DNA by neutrophils.

NETosis Initiation:

 Receptors such as TNF, Fc, and GPCR, as well as CD18, play a role in NETosis
initiation in response to various triggers. Inflammation is important

 Calcium spikes, which mediate enzymes like PAD4, are crucial for NETosis
regulation. And will be discussed later

 Kinases (Janus kinase 2, Raf-MEK-ERK MAP kinase pathway, etc.) and

reactive oxygen species (ROS) also impact NETosis.

Mechanism of DNA Decondensation:

Chromatin decondensation in NETosis involves histone posttranslational alterations,

including citrullination and cleavage mediated by serine proteases and PAD4.

 PAD4 and Histone Citrullination:

PAD4 promotes NETosis by citrullinating histones, leading to chromatin

decondensation.
thereby affecting transcription control by altering histone-DNA interactions.

PAD4 triggers histone detachment during NETosis, causing chromatin loss.

Elevated in chronic illnesses and sterile inflammation.

 Role of Proteases:

Primary granule-resident proteases (PR3, cathepsin G, neutrophil elastase)

are essential for chromatin decondensation and NETosis.

Proteases, such as PR3 and neutrophil elastase, are crucial for DNA liberation
from histones during chromatin decondensation and NETosis, and their entry
into the nucleus unknown.
 Role of Myeloperoxidase:

Myeloperoxidase enhances chromatin decondensation, and its role in NET

release is under investigation.
DNA Release from Nucleus to Cytosol:

DNA release from the nucleus involves lamin remodelling and nuclear envelope hole
development, separating chromatin from the cytoplasm and allowing molecular
shock absorption and expression.

Lamin dynamics and nuclear membrane dynamics play key roles in DNA release
during NETosis.

 NETosis modifications to lamin meshwork result in decondensed DNA

extrusion and small discontinuities in lamin B1+2 networks, possibly due to
lamin posttranslational changes and PAD4.

 NETosis and apoptosis involve nuclear membrane vesiculation, releasing

chromatin into the cytoplasm. The collapse of the cytoskeleton causes
neutrophils to rupture their nuclear envelope, possibly through gasdermin D-
mediated permeabilization.

Remodeling of Cytoskeletal and Membranous Organelles:

Disassembling actin filaments and MTs during NETosis is a major remodeling

process before extracellular DNA release.

 Actin disintegration is crucial for NETosis, stimulate extracellular trap

formation. Causes usually include increase intracellular calcium
concentration, actin oxidation, neutrophil elastase translocation, and actin
depolymerization.

 The cytosol undergoes major alterations during NETosis, and minutes

after the cytoskeleton disassembles, the ER vesiculates. On NETs, neutrophil
elastase and MPO are found. ROS generation is mediated by mitochondria
and is required for the release of mitochondrial DNA and calcium ionophores.
.

Extracellular DNA Release:

 Multiple stages in NETosis involve a rise in plasma membrane permeability

before extracellular DNA release.
 Pore-forming proteins like gasdermin D are expressed by neutrophils during
NETosis.
Conclusion:

NETosis is a complex process involving various cellular components and pathways.

Understanding the molecular mechanisms of NETosis is crucial for developing

effective treatment strategies.