# Impact of Neutrophil Extracellular Traps (NETs) on the Pancreatic Tumor Microenvironment

> **NIH NIH P20** · WEST VIRGINIA UNIVERSITY · 2021 · $246,840

## Abstract

Pancreatic adenocarcinoma is a devastating malignancy characterized by a uniquely fibrotic pancreatic 
tumor stroma that limits immune cell infiltration, promotes tumor growth through cancer-stromal signaling 
pathways and limits delivery of systemic treatments. Development of novel therapeutic strategies that target 
the pathophysiology of pancreatic cancer and the characteristic fibrosis associated with it are desperately 
needed. Recently, the process of neutrophil extracellular traps (NETs), in which activated neutrophils release 
their intracellular contents including DNA, histones, and granules into the extracellular tissue or circulation, 
have been implicated in pancreatic cancer. Protein arginine deiminase 4 (PAD4) is an enzyme that 
citrullinates histones to allow for unwinding and expulsion from the cell and is required for NET formation, 
providing a potential therapeutic target for NET inhibition in cancer. PAD4-/- mice have diminished local and 
systemic NET formation, resulting in limited tumor growth and improved survival in murine pancreatic cancer. 
In addition to promoting pancreatic tumor growth, NETs also contribute to the spread of metastatic disease 
and cancer-associated hypercoagulability. We have demonstrated that neutrophil DNA released from NETs 
also activate pancreatic stellate cells (PSCs), the principle cell responsible for fibrosis in the pancreatic tumor 
microenvironment (TME). Our overall objective in the current proposal is to identify the impact of NETs on 
hypoxia, acidosis and metabolism in the pancreatic TME, and target NETs with pharmacologic inhibitors. In 
Aim 1, we will utilize innovative electron paramagnetic resonance (EPR) to provide in vivo assessment of 
oxygen content and pH in the pancreatic TME. We will compare tumors from wild type and PAD4-/- mice, as 
well as mice treated with NET inhibitors to determine the influence of NETs on these parameters. NETs 
release damage associated pattern molecules (DAMPs), which are known to influence metabolic processes 
such as mitochondrial function and autophagy. In Aim 2, we will utilize the Seahorse assay to determine 
how NETs alter cell metabolism in primary pancreatic cancer cells, pancreatic stellate cells and whole tumors. 
We will also explore how NETs activate PSCs and increase fibrosis in the TME. These studies are critically 
important to improving our understanding of the pancreatic tumor microenvironment. This work will provide 
preliminary data in support of future grants funding mechanistic studies and target these processes at a time 
when novel therapies are desperately needed for this devastating disease.

## Key facts

- **NIH application ID:** 10485123
- **Project number:** 5P20GM121322-04
- **Recipient organization:** WEST VIRGINIA UNIVERSITY
- **Principal Investigator:** BRIAN A BOONE
- **Activity code:** P20 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $246,840
- **Award type:** 5
- **Project period:** 2021-07-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10485123

## Citation

> US National Institutes of Health, RePORTER application 10485123, Impact of Neutrophil Extracellular Traps (NETs) on the Pancreatic Tumor Microenvironment (5P20GM121322-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10485123. Licensed CC0.

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