PB1417 - Pleiotropic roles of FVIII: new insight in endothelial cell functionality

Tuesday, June 27, 2023

18:30 – 19:30 ET

Room: Exhibition

Presenting Author(s)

Alessia Cucci, PhD (she/her/hers)

Postdoc
Department of Health Sciences, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy

Co-Author(s)

Cristina Olgasi, PhD

Tenure-track Assistant Professor
Department of of Translational Medicine, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy
IM

Ivan Molineris

Associate Professor
Italian Institute for Genomic Medicine (IIGM); Candiolo, Italy
Turin, Piemonte, Italy
SA

Simone Assanelli

PhD student
Department of Health Sciences, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy
FA

Francesca Anselmi

Technician
Italian Institute for Genomic Medicine (IIGM); Candiolo, Italy
Turin, Piemonte, Italy
CB

Chiara Borsotti

Tenure-track Assistant Professor
Department of Health Sciences, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy
CS

Chiara Sgromo

PhD student
Department of Health Sciences, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy
AL

Andrea Lauria

Research Assistant Professor
University of Turin, Turin, Italy
Turin, Piemonte, Italy
SM

Simone Merlin

Associate Professor
Department of Health Sciences, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy
GW

Gillian Walker

Technologist
Department of Health Sciences, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy
PC

Paola Capasso

Technician
San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), Milan, Italy
Milan, Piemonte, Italy
AL

Angelo Lombardo

Associate Professor
San Raffaele Telethon Institute for Gene Therapy (SR-Tiget), Milan, Italy
Milan, Piemonte, Italy
SO

Salvatore Oliviero

Full professor
Italian Institute for Genomic Medicine (IIGM); Candiolo, Italy
Turin, Piemonte, Italy
Antonia Follenzi, MD, PhD

Full professor
Department of Health Sciences, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy

Background: Hemophilia A (HA) is a X-linked bleeding disorder caused by mutations of coagulation factor VIII (FVIII). Main clinical manifestations are spontaneous bleedings that primarily consist of hemarthrosis and intracranial hemorrhages. To date, the impairment of vessel stability in HA patients and the correlation between FVIII and endothelial functionality are poorly understood.

Aims: Therefore, we aimed to investigate the extra-coagulative role of FVIII in the maintenance of endothelial cells (ECs) functionality by studying the transcriptomic significant differences in HA and healthy ECs.

Methods: Blood Outgrowth ECs (BOECs) from HA patients and healthy donors were used. HA BOECs were transduced with a lentiviral vector (LV) carrying the B-domain deleted FVIII (LV-FVIII). The transcriptomic profile of healthy, HA and LV-FVIII HA BOECs was evaluated by RNA-seq technology. Healthy ECs, HA ECs, LV-FVIII ECs, as well as KO-F8 ECs generated by CRISPR/Cas9 technology, were used to investigate the role of FVIII in ECs functionality by tubulogenesis, migration, and permeability assays both in vitro and in vivo in a mouse model of HA.

Results: BOECs transcriptomic profiles revealed that FVIII regulates cell adhesion, cell migration and extracellular matrix organization genes. HA ECs, as well as KO-F8 ECs, showed an alteration of vessel-formation, migration, and permeability compared to controls. Importantly, the impaired EC phenotype was rescued by treatment with recombinant human FVIII or by LV expressing FVIII as confirmed by the upregulation of related genes. Finally, the endothelial function of FVIII was confirmed in vivo in a mouse model of severe HA which showed that an altered angiogenesis and vessel permeability could be corrected by exogenous FVIII.

Conclusion(s): The downmodulated genes found in FVIII deficient HA BOEC can explain their impaired EC functionality. These results provide insights for novel FVIII roles that could be instrumental for therapeutic strategies to treat HA patients.