PB1369 - Neutrophil extracellular traps as orchestrators of bladder cancer progression: Unveiling their protumorigenic mechanisms and their effect on coagulation

Tuesday, June 25, 2024

13:45 – 14:45 ICT

Room: Exhibition Hall

Co-Author(s)

RH

Raquel Herranz

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

Patricia Ibiza-Guijarro

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

María Castaño

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

Isabel Alemán-Cabrera

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

Presenting Author(s)

JO

Julia Oto

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

Co-Author(s)

PV

Patricia Verger

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

Fernando Cana

Laboratory technician
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, 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, Comunidad Valenciana, Spain
PM

Pau Martínez-Rodríguez

Medical doctor
Urology Service, Manises Hospital
Valencia, Comunidad Valenciana, Spain
CV

César David Vera-Donoso

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

Manuel Martínez-Sarmiento

Medical doctor
Urology Service, La Fe University and Polytechnic Hospital
Valencia, Comunidad Valenciana, Spain
Pilar Medina, PhD

Principal Investigator
Haemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Health Research Institute Hospital La Fe
Valencia, Comunidad Valenciana, Spain

Background: We previously evidenced an increased release of neutrophil extracellular traps (NETs) in bladder cancer (BC) patients systemically and in the tumor microenvironment. These seem to enhance tumor growth and metastasis, but the mechanism remains unknown.

Aims: To evaluate, whether NETs affect BC hallmarks, and whether this effect differs in NETs derived from BC patients compared to those obtained from healthy individuals.

Methods: Blood neutrophils were purified from 4 BC patients and 4 healthy controls and NETs were obtained by induction with 500nM PMA. Two BC cell lines (grade 3-T24 and grade 2-5637, ATCC) were treated with increasing NETs concentrations to evaluate differences in cell proliferation (sodium phosphatase cell viability assay), migration (scratch assay), extracellular matrix invasion (transwell assay) and Anoikis resistance. To evaluate whether NETs induce the release of procoagulant molecules by the BC cells, we performed the thrombin generation test (TGT, Stago) in a pool of plasma with the post treatment conditioned media (CM) and subtracted the effect of the treating NETs on each parameter.

Results: High concentration of NETs (500ng/ml) increased cell proliferation and extracellular matrix invasion of the T24 cells, more significantly in BC-derived NETs than in NETs derived from healthy controls. NETs also decreased T24 migration (Figure1). NETs didn´t modify the behaviour of 5637 cells.
Compared to a pool of plasma, untreated T24 CM accelerated thrombin generation but produced lower thrombin levels (P < 0.01). NETs treatment increased the thrombin peak and normalized the speed of the reaction (P < 0.05) (Figure2).

Conclusion(s): NETs affect high-grade BC cells´ behavior by increasing proliferation and invasion and reducing migration, which could help cancer progression and metastasis implantation. This effect is more significant with BC patient-derived NETs. Moreover, NETs induce T24 cells to release molecules that increase thrombin generation, possibly modifying the procoagulant potential of BC cells. ISCIII-FEDER (PI20/00075, FI21/00171, PI23/00449) and SETH.