Our preliminary data indicate that molecularly engineered BOEC (blood outgrowth endothelial cells) can be used for effective gene therapy of hemophilia A. The present project will examine several aspects of BOEC biology that are directly relevant to this potential therapeutic use of BOEC.
Three Specific Aims will utilize the model of human BOEC (either unmanipulated or engineered to express human factor VIII) given to NOD. SCID mice.
In Aim 1 we will identify the marrow """"""""homing"""""""" behavior of BOEC. We will determine where BOEC go in the short- and long-terms when given intravenously. We will identify mechanisms underlying BOEC seeding of marrow and spleen. We will determine if BOEC seeding of marrow/spleen can be increase pharmacologically (with histamine, or beta-1-integrin activating antibody, or VEGF, or 1L6 plus stem cell factor). We will determine if the apparently low seeding frequency of BOEC in marrow is a stochastic or pre-programmed characteristic of establishing a BOEC graft.
In Aim 2 we will examine certain issues regarding BOEC graft expansion and longevity in vivo. We will confirm our preliminary impression that BOEC expand substantially in vivo after being administered intravenously. We will determine if this in vivo expansion is predicatble and is dependent on the prior passaging history of the BOEC. We will seek to confirm that silencing of our fVIlI transgene expression vector is not a problem in BOEC.
in Aim 3 we expect to document that endothelial expansion in vivo after intravenous administration of BOEC is due to a true BOEC cell rather than a contaminating hematopoietic precursor cell. Each of these studies addresses an issue that is necessary to understand before BOEC technology can be applied for therapeutics. Aside from eventual therapeutics, these studies will define aspects of this unique cell type.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
Project #
Application #
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Link, Rebecca P
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Minnesota Twin Cities
Internal Medicine/Medicine
Schools of Medicine
United States
Zip Code
Milbauer, Liming C; Enenstein, Judy A; Roney, Mark et al. (2009) Blood outgrowth endothelial cell migration and trapping in vivo: a window into gene therapy. Transl Res 153:179-89
Matsui, Hideto; Shibata, Masaru; Brown, Brian et al. (2007) Ex vivo gene therapy for hemophilia A that enhances safe delivery and sustained in vivo factor VIII expression from lentivirally engineered endothelial progenitors. Stem Cells 25:2660-9
Somani, Arif; Nguyen, Julia; Milbauer, Liming C et al. (2007) The establishment of murine blood outgrowth endothelial cells and observations relevant to gene therapy. Transl Res 150:30-9
De Meyer, Simon F; Vanhoorelbeke, Karen; Chuah, Marinee K et al. (2006) Phenotypic correction of von Willebrand disease type 3 blood-derived endothelial cells with lentiviral vectors expressing von Willebrand factor. Blood 107:4728-36
Stachelek, Stanley J; Alferiev, Ivan; Choi, Hoon et al. (2005) Cholesterol-derivatized polyurethane: characterization and endothelial cell adhesion. J Biomed Mater Res A 72:200-12
Fernandez-L, Africa; Sanz-Rodriguez, Francisco; Zarrabeitia, Roberto et al. (2005) Blood outgrowth endothelial cells from Hereditary Haemorrhagic Telangiectasia patients reveal abnormalities compatible with vascular lesions. Cardiovasc Res 68:235-48
Sieminski, A L; Hebbel, R P; Gooch, K J (2005) Improved microvascular network in vitro by human blood outgrowth endothelial cells relative to vessel-derived endothelial cells. Tissue Eng 11:1332-45
Sieminski, A L; Hebbel, R P; Gooch, K J (2004) The relative magnitudes of endothelial force generation and matrix stiffness modulate capillary morphogenesis in vitro. Exp Cell Res 297:574-84