Vascular Biology
Cristina Olgasi, PhD
Tenure-track Assistant Professor
Department of of Translational Medicine, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy
Alessia Cucci
post doc
Department of Health Sciences, Università del Piemonte Orientale
Novara, Piemonte, Italy
Simone Assanelli
phD student
Department of Health Sciences, Università del Piemonte Orientale
Novara, Piemonte, Italy
Chiara Sgromo
phD student
Department of Health Sciences, Università del Piemonte Orientale
Novara, Piemonte, Italy
Chiara Borsotti
Assistant professor
Department of Health Sciences, Università del Piemonte Orientale
Novara, Piemonte, Italy
Gillian Walker
Technologist
Department of Health Sciences, University of Piemonte Orientale, Novara, Italy
Novara, Piemonte, Italy
Ivan Molineris
Associate Professor
Italian Institute for Genomic Medicine (IIGM); Candiolo, Italy
Turin, Piemonte, Italy
Francesca Anselmi
Technician
Italian Institute for Genomic Medicine (IIGM); Candiolo, Italy
Turin, Piemonte, Italy
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
Hemophilia A (HA) is a rare bleeding disorder caused by the absence or dysfunction of Factor FVIII (FVIII). Clinical manifestations are spontaneous bleedings that primarily consist of hemarthroses and intracranial hemorrhages. Standard therapies are ineffective in preventing the bleeding episodes and they can occur without any clear cause. To date, the impairment of vessel stability in HA patients and a correlation between FVIII and endothelial functionality has never been explained.
Aims:
To elucidate the potential role of FVIII in endothelial stability and investigate significant differences in HA and healthy endothelial cells (ECs).
Methods:
iPSCs-derived ECs and Blood Outgrowth ECs (BOECs), both from HA patients and healthy donors, were used as ECs models. HA-ECs were transduced with a lentiviral vector (LV) carrying the B-deleted form of FVIII under the control of an endothelial specific-promoter (LV-VEC.FVIII). The transcriptomic profile of healthy, HA, and LV-VEC.FVIII-transduced HA ECs were evaluated by RNA Seq analysis and the differences in EC functionality were analyzed both in vitro and in vivo in a HA mouse model.
Results:
Transcriptomic analysis revealed different gene expression in HA vs healthy ECs in both ECs models. The impaired phenotype was partially attenuated in LV-VEC.FVIII transduced HA ECs. Several genes were down regulated in HA ECs compared to healthy ECs, suggesting an impairment in HA ECs stability reproducible in both ECs systems. These data were validated in vitro, showing an impaired vessel-formation capability, migration potential, and permeability for HA ECs. Finally, in a mouse model of severe HA, it was demonstrated an altered permeability and tubulogenesis potential of HA vessels when compared to wild-type mice.
Conclusion(s):
These results as confirmed in our EC models provide new insights into unexplored roles for FVIII besides coagulation, offering new therapeutic gene and cell therapy strategies in the management of HA patients.