# Treatment of Deep Vein Thrombosis via Targeted Inhibition of the FXII-uPAR-pAkt2 Axis in Neutrophils

> **NIH VA I01** · LOUIS STOKES CLEVELAND VA MEDICAL CENTER · 2021 · —

## Abstract

The overall goal of this proposal is to establish a targeted therapeutic strategy to disrupt the interaction of
coagulation factor FXII (FXII) and urokinase plasminogen activator receptor (uPAR) to downregulate Akt2-
mediated neutrophil activation for treatment of deep vein thrombosis (DVT). DVT is a leading cause of
cardiovascular death. New anticoagulation therapies have been developed, however these therapeutic
advances are all associated with increased rate of bleeding. Moreover, these current anticoagulants only inhibit
coagulation end-points (e.g. thrombin and fibrin) but do not prevent upstream events such as neutrophil
activation, procoagulant neutrophil extracellular trap (NET) formation, or propagation of neutrophil-platelet
aggregates, all of which are persistent hallmark events in DVT. In this framework, we propose to use a unique
nanomedicine-based therapeutic approach to downregulate neutrophil activation and NET formation through
targeted disruption of the FXII-uPAR-pAkt2 axis. Our laboratory identified that FXII in neutrophils is critical for
function. Specifically, we have shown that following neutrophil activation, autocrine FXII signals through uPAR
leading to Akt2S474 phosphorylation (pAkt2) and NET formation. Inhibiting FXII signaling in neutrophils resulted
in smaller venous thrombi. Based on these mechanistic findings, our central hypothesis is that targeted inhibition
of the FXII-uPAR-pAkt2 axis will be therapeutically effective in treating DVT while minimizing systemic side-
effects and bleeding risk. We will test this hypothesis by packaging uPAR inhibitory peptides within nanovesicles
that are uniquely surface-engineered to undergo specific heteromultivalent anchorage onto neutrophil-platelet
aggregates.
In this application, our goals are to: 1) identify a candidate uPAR inhibitory peptide that disrupts FXII binding on
the surface of neutrophils. We will determine the affinity, stoichiometry and specificity of inhibition, assess Akt2
activation and perform neutrophil and platelet function assays; 2) use heteromultivalently decorated nanovesicles
loaded with the candidate peptide drug to determine their ability to site-selectively inhibit the FXII-uPAR
interaction and mitigate DVT in vitro and in vivo; 3) to validate these preclinical studies, we will determine the
constitutive activity of the FXII-uPAR-pAkt2 axis and the effect of its inhibition on neutrophil functions, neutrophil-
platelet interactions and thrombus growth ex vivo, using blood samples from patients with newly diagnosed DVT.
The end goal is to show the differential abundance of the FXII-uPAR-pAkt2 axis and downstream effectors in
DVT pathology which will lay the foundation for future clinical studies to inhibit its action.
Our scientific innovation is the mechanistic elucidation of the FXII-uPAR-pAkt2 signaling axis in neutrophil-
mediated pathology. Our technological innovation is the development of inhibitory peptide-based targeted
nanomedicine strateg...

## Key facts

- **NIH application ID:** 10044407
- **Project number:** 5I01BX003851-02
- **Recipient organization:** LOUIS STOKES CLEVELAND VA MEDICAL CENTER
- **Principal Investigator:** Evi X. Stavrou
- **Activity code:** I01 (R01, R21, SBIR, etc.)
- **Funding institute:** VA
- **Fiscal year:** 2021
- **Award amount:** —
- **Award type:** 5
- **Project period:** 2019-10-01 → 2023-09-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10044407

## Citation

> US National Institutes of Health, RePORTER application 10044407, Treatment of Deep Vein Thrombosis via Targeted Inhibition of the FXII-uPAR-pAkt2 Axis in Neutrophils (5I01BX003851-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10044407. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*