PB0783 - Suramin is a Hepsin inhibitor which reduces its protumor and prothrombotic effects in colorectal cancer cells

Monday, July 11, 2022

6:30 PM – 7:30 PM

Location: Exhibition

Presentation Format: In-Person

Presenting Author(s)

David Zaragoza Huesca, PhD student

PhD student
Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla
Murcia, Murcia, Spain

Co-Author(s)

MR

Mari Carmen Ródenas

PhD
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain
Murcia, Murcia, Spain
JP

Julia Peñas-Martínez

PhD student
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain
Murcia, Murcia, Spain
IP

Irene Pardo Sánchez

PhD student
Department of Cell Biology, Faculty of Biology, Universidad de Murcia. CIBERER, IMIB-Arrixaca, Murcia
Murcia, Murcia, Spain
CO

Carmen Ortega Sabater

PhD student
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
murcia, Murcia, Spain
JP

Jorge Peña García

PhD
Structural Bioinformatics and High Performance Computing Research Group (BIOHPC), Computer Engineering Department, UCAM Universidad Católica de Murcia, 30107 Guadalupe, Spain
Murcia, Murcia, Spain
SE

Salvador Espín

Technician
Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla
Murcia, Murcia, Spain
GR

Guillermo Ricote Sánchez

Medical Doctor
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain
Murcia, Murcia, Spain
SM

Sofía Montenegro Luis

Medical Doctor
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain
Murcia, Murcia, Spain
FA

Francisco Ayala de la peña

Medical Doctor
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain
Murcia, Murcia, Spain
GL

Ginés Luengo Gil

PhD
Clinical Analysis and Pathology Department, Group of Molecular Pathology and Pharmacogenetics, IMIB-Arrixaca, Hospital Universitario Santa Lucía, 30202 Cartagena, Spain.
Murcia, Murcia, Spain
AN

Andrés Nieto Olivares

PhD
Department of Pathology, Hospital Universitario Morales Meseguer, Murcia, Spain.
Murcia, Murcia, Spain
FG

Francisco García Molina

PhD
Department of Pathology, Hospital Universitario Reina Sofía, Murcia, Spain.
Murcia, Murcia, Spain
VV

Vicente Vicente

Researcher and Medical Doctor
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, CIBERER, Murcia, Spain
Murcia, Murcia, Spain
FB

Francesco Bernardi

Professor of Biochemistry
Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, Ferrara University, Ferrara (Italy)
Verona, Veneto, Italy
VM

Victoriano Mulero

PhD
Department of Cell Biology, Faculty of Biology, Universidad de Murcia. CIBERER, IMIB-Arrixaca, Murcia
Murcia, Murcia, Spain
HP

Horacio Pérez-Sánchez

Professor
Structural Bioinformatics & High Performance Computing Research Group (BIO-HPC), Universidad Católica de Murcia (UCAM), Murcia, 30107, Spain
Murcia, Murcia, Spain
AC

Alberto Carmona Bayonas

Medical Doctor
Department of Hematology and Medical Oncology, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, CIBERER, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
Murcia, Murcia, Spain
Irene Martínez Martínez, PhD

Principal Investigator
Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla
Murcia, Murcia, Spain

Background: Hepsin (HPN) is an extracellular serine-protease which dysregulation leads to tumor invasion and malignancy. Our group previously described HPN increases protumor and prothrombotic phenotypes in colorectal cancer (CRC) cells, and promotes invasion in zebrafish models. We also identified higher HPN plasma levels in metastatic vs. localized CRC patients, and in patients with thrombosis vs. no thrombosis.

Aims: To identify new HPN inhibitors that reduced its protumor and prothrombotic effects in CRC cells.

Methods: We performed a virtual screening to identify potential compounds that blocked HPN active site. The most outstanding compounds were tested in a fluorogenic assay that measured HPN proteolytic activity. Then, we measured HPN inhibitors effect on CRC cell line Caco-2 transfected with HPN-expression vector (Caco-2-HPN; HPN-overexpression) and negative-control vector (Caco-2; HPN-endogenous expression). We conducted wound confluence, gelatin matrix, cell cytometry and thrombin generation assays (TGA) for measuring migration, invasion, proliferation and procoagulant cell phenotypes, respectively. Finally, zebrafish models were used to test whether HPN inhibitors interfered with distant tumor dissemination.

Results: Virtual screening highlighted suramin as the most effective HPN inhibitor, with a docking score= -12 Kcal/mol. Fluorogenic assay confirmed suramin inhibited HPN in an irreversible and dose-dependent way (IC50=0.66 µM; 95%IC=0.42-1.32). In Caco-2 and Caco-2-HPN cells, suramin reduced migration and, significantly, invasion phenotypes (Figure 1A,B). These effects were more pronounced in Caco-2-HPN cells. Also in both cell types, suramin increased lag time and reduced endogenous thrombin potential and thrombin peak in TGA. Moreover, suramin increased time to peak and reduced velocity to peak in Caco-2-HPN cells (Figure 2A). Finally, suramin reduced Caco-2-HPN invasion to Caco-2 levels in zebrafish (Figure 2B).

Conclusion(s): Suramin is a HPN inhibitor that reduces invasive, migratory and procoagulant characteristics of CRC cells in vitro. In addition, suramin reduces the invasion phenotype in a zebrafish model. These results point suramin as a potential therapeutic agent in CRC.