PB0728 - Bladder cancer patients have increased NETosis and impaired DNaseI-mediated NET degradation that can be therapeutically restored in vitro

Monday, July 11, 2022

6:30 PM – 7:30 PM

Location: Exhibition

Presentation Format: In-Person

Presenting Author(s)

Raquel Herranz, MSc

PhD Candidate
Hemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain

Co-Author(s)

Julia Oto

Postdoctoral Researcher
Hemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
MH

Marta Hueso

Researcher
Haemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe
Valencia, Comunidad Valenciana, Spain
EP

Emma Plana

Postdoctoral Researcher
Haemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain; Angiology and Vascular Surgery Service, La Fe University and Polytechnic Hospital, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
FC

Fernando Cana

Laboratory Technician
Hemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
AF

Alvaro Fernández-Pardo

Researcher
Haemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe
Valencia, Comunidad Valenciana, Spain
SB

Santiago Bonanad

Head of Thrombosis and Hemostasis Unit
Haemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain; Thrombosis and Haemostasis Unit, Haematology Service, La Fe University and Polytechnic Hospital, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
CV

Cesar D. Vera-Donoso

Medical doctor
Department of Urology, La Fe University and Polytechnic Hospital, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
MM

Manuel Martínez-Sarmiento

Medical doctor
Department of Urology, La Fe University and Polytechnic Hospital, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
PM

Pilar Medina

PI
Hemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain

Background: An increased release of neutrophil extracellular traps (NETs), through a process called NETosis, occurs in cancer. This enhances tumor growth and metastasis, which may be mediated by impaired NET degradation. However, this has never been evidenced in bladder cancer (BC).

Aims: To evaluate the increase in NETosis in plasma of BC patients, to ascertain whether it is mediated by DNaseI and to in vitro explore therapeutic interventions.
To compare the ability of plasma from patients with BC and from healthy volunteers (controls) to degrade NETs. Also, we aimed to evaluate the effect of the addition of Dornase Alfa (a recombinant human DNaseI used in clinical practice for management and treatment of cystic fibrosis) in NET degradation.

Methods: NETs markers were measured in plasma samples of 25 BC patients and 23 controls. Plasma DNaseI activity was measured with the SRED assay. Neutrophils were isolated from a healthy volunteer, seeded and activated with phorbol myristate acetate (PMA) to prompt NETosis. NETs were incubated with 5 µl plasma from each participant. In parallel, plasma from BC patients was supplemented with Dornase Alfa (Pulmozyme, Roche), control pooled plasma or vehicle (PBS 1X + 0.1%BSA).

Results: BC patients had significantly increased NETs markers in plasma compared to controls (P < 0.0001) and lower DNaseI activity (P < 0.0001) (Figure 1). In vitro NET degradation was impaired when incubated with BC plasma compared to control plasma (P < 0.0001) (Figure 2). Addition of DNaseI restored NET degradation up to the level of healthy controls (P < 0.0001) (Figure 2).

Conclusion(s): BC patients have increased NETosis in part mediated by a reduced DNaseI activity. In vitro addition of exogenous DNaseI, an approved treatment for cystic fibrosis, restores NET degradation in BC patients, thus becoming a potential therapeutic tool to reduce the risk of cancer-associated thrombosis and metastasis.
ISCIII-FEDER (PI17/00495, PI20/00075, FI21/00171), GVA (ACIF/2017/138) and SETH.