Faculty Collaboration Database

Genetic induction of a releasable pool of factor VIII in human endothelial cells. Arterioscler Thromb Vasc Biol 2000 Dec;20(12):2689-95

Date

12/16/2000

Pubmed ID

11116073

DOI

10.1161/01.atv.20.12.2689

Scopus ID

2-s2.0-0033669966 (requires institutional sign-in at Scopus site)   72 Citations

Abstract

Although it is known that factor VIII (FVIII) plasma levels increase rapidly in response to a number of stimuli, the biological stimuli behind this release is less clear. Previously, we showed that FVIII can traffic together with von Willebrand factor (vWF) into storage granules in a pituitary tumor cell line, AtT-20; however, AtT-20 cells could not be used to address the release or functional activity of released FVIII. To investigate the regulated secretion of stored FVIII, endothelial cells with intact agonist-stimulated release pathways were used. Human umbilical vein endothelial cells (HUVECs) were transduced with retroviral FVIII construct [hFVIII(V)] to create a FVIII/vWF storage pool. Immunofluorescent staining of transduced cells demonstrated FVIII in Weibel-Palade bodies. In contrast, the transduction of hFVIII(V) into HT-1080 and HepG2 cells displayed FVIII only in the cytoplasm. We studied the regulated release of both FVIII and vWF from endothelial cells after agonist-induced stimulation and demonstrated a parallel release of FVIII and vWF proteins. This released FVIII was functionally active. Hence, endothelial cells transduced with hFVIII(V) store FVIII together with vWF in Weibel-Palade bodies, creating a releasable storage pool of both proteins. Because FVIII secretion can be physiologically regulated by agonists in culture, this may explain the pharmacological agonist-induced release of FVIII by drugs such as desmopressin in vivo and suggests vascular endothelium as a reasonable target of gene therapy of hemophilia A.

Author List

Rosenberg JB, Greengard JS, Montgomery RR

MESH terms used to index this publication - Major topics in bold

Cell Line
Cells, Cultured
Cytoplasm
Deamino Arginine Vasopressin
Endothelium, Vascular
Factor VIII
Fluorescent Antibody Technique, Indirect
Genetic Therapy
Genetic Vectors
Hemophilia A
Hemostatics
Humans
Transduction, Genetic
von Willebrand Factor