Extrahepatic sources of factor VIII potentially contribute to the coagulation cascade correcting the bleeding phenotype of mice with hemophilia A.
A large fraction of factor VIII in blood originates from liver sinusoidal endothelial cells although extrahepatic sources also contribute to plasma factor VIII levels. Identification of cell-types other than endothelial cells with the capacity to synthesize and release factor VIII will be helpful for therapeutic approaches in hemophilia A. Recent cell therapy and bone marrow transplantation studies indicated that Küpffer cells, monocytes and mesenchymal stromal cells could synthesize factor VIII in sufficient amount to ameliorate the bleeding phenotype in hemophilic mice. To further establish the role of blood cells in expressing factor VIII, we studied various types of mouse and human hematopoietic cells. We identified factor VIII in cells isolated from peripheral and cord blood, as well as bone marrow. Co-staining for cell type-specific markers verified that factor VIII was expressed in monocytes, macrophages and megakaryocytes. We additionally verified that factor VIII was expressed in liver sinusoidal endothelial cells and endothelial cells elsewhere, e.g., in the spleen, lungs and kidneys. Factor VIII was well expressed in sinusoidal endothelial cells and Küpffer cells isolated from human liver, whereas by comparison isolated human hepatocytes expressed factor VIII at very low levels. After transplantation of CD34(+) human cord blood cells into NOD/SCIDγNull-hemophilia A mice, fluorescence activated cell sorting of peripheral blood showed >40% donor cells engrafted in the majority of mice. In these animals, plasma factor VIII activity 12 weeks after cell transplantation was up to 5% and nine of 12 mice survived after a tail clip-assay. In conclusion, hematopoietic cells, in addition to endothelial cells, express and secrete factor VIII: this information should offer further opportunities for understanding mechanisms of factor VIII synthesis and replenishment.
Dept. of Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
Diego
D
Zanolini
Dept. of Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
Simone
S
Merlin
Dept. of Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
Maria
M
Feola
Dept. of Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
Gabriella
G
Ranaldo
Dept. of Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
Angela
A
Amoruso
Dept. of Translational Medicine, Università del Piemonte Orientale, Novara, Italy.
Gianluca
G
Gaidano
Azienda Ospedaliera Universitaria Maggiore della Carità, SCDU Pediatria, Novara, Italy.
Mauro
M
Zaffaroni
Azienda Ospedaliera Mauriziano, SC Chirurgia Generale ed Oncologica, Torino, Italy.
Alessandro
A
Ferrero
Dept. of Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
Sandra
S
Brunelleschi
Dept. of Translational Medicine, Università del Piemonte Orientale, Novara, Italy.
Guido
G
Valente
Dept. of Medicine and Pathology, Marion Bessin Liver Research Center, Diabetes Center, Cancer Center, and Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, Bronx, NY, USA.
Sanjeev
S
Gupta
Dept. of Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
Maria
M
Prat
Dept. of Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy antonia.follenzi@med.uniupo.it.
Antonia
A
Follenzi
D000818:Animals; D001777:Blood Coagulation / Q000235:genetics; D036101:Cord Blood Stem Cell Transplantation; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D005169:Factor VIII / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation; D006085:Graft Survival; D006467:Hemophilia A / Q000097:blood / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D006470:Hemorrhage / Q000097:blood / Q000235:genetics / Q000517:prevention & control*; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007728:Kupffer Cells / Q000378:metabolism* / Q000473:pathology; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008533:Megakaryocytes / Q000378:metabolism / Q000473:pathology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009000:Monocytes / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D014183:Transplantation, Heterologous
D005169:Factor VIII