# Differential targeting of von Willebrand factor depending on oxidizing conditions

> **NIH NIH R01** · UNIVERSITY OF WASHINGTON · 2022 · $406,738

## Abstract

Project Summary / Abstract
The blood protein von Willebrand factor (VWF) is a large multimeric protein that, when activated, binds to
blood platelets tethering them to the site of vascular injury initiating blood coagulation. This process is critical
for normal haemostasis, but especially under inflammatory conditions it is thought to be a major player in patho-
logical thrombus formation. For this reason, VWF has been the target for the development of anti-thrombotic
therapeutics. However, a particular challenge is how to prevent pathological thrombus formation while still allow-
ing normal physiological blood coagulation. In fact, currently available anti-thrombotic therapeutics are known
to cause intracranial bleeding as side effect. The work presented here proposes that by identifying the confor-
mational changes that VWF undergoes in inflammation it will be possible to design molecules that target VWF
selectively only in an inflammatory pro-thrombotic environment. Experiments performed in vitro have indicated
that oxidizing agents released during inflammation increase the platelet-binding activity of VWF, and this has
been linked to the oxidation of methionine residues within VWF. This study aims to characterize what inhibitory
mechanisms are removed in VWF through methionine oxidation and identifies sites that can be targeted optimally
under oxidizing conditions. We hypothesize that oxidation activates the A1 domain (the domain in VWF that
contains the binding site to platelets) by removing the masking function of its neighboring domains. Through
a combination of a dynamic flow assay, equilibrium unfolding experiments and a binding assay, we will identify
which auto-inhibitory mechanisms of VWF are turned off by oxidation and which methionine residues are key for
activation under oxidizing conditions. This knowledge will then be used in computational docking studies to create
a model of the complex between the A1 domain and its neighboring domains and study the structural effects of
methionine oxidation at atomic level of detail. An assay using samples from patients with inflammatory conditions
will be used to test the link between inflammatory conditions and higher activity of VWF in vivo. The structural
insights gained here will be invaluable to structure-based drug design in order to develop therapeutics that target
VWF only when it is in its oxidized state in order to prevent thrombosis while allowing haemostasis.

## Key facts

- **NIH application ID:** 10474452
- **Project number:** 5R01HL153253-02
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Gianluca Interlandi
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $406,738
- **Award type:** 5
- **Project period:** 2021-09-01 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10474452

## Citation

> US National Institutes of Health, RePORTER application 10474452, Differential targeting of von Willebrand factor depending on oxidizing conditions (5R01HL153253-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10474452. Licensed CC0.

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