Adenovirus-mediated BMP2 expression in human bone marrow stromal cells.
Recombinant adenoviral vectors have been shown to be potential new tools for a variety of musculoskeletal defects. Much emphasis in the field of orthopedic research has been placed on developing systems for the production of bone. This study aims to determine the necessary conditions for sustained production of high levels of active bone morphogenetic protein 2 (BMP2) using a recombinant adenovirus type 5 (Ad5BMP2) capable of eliciting BMP2 synthesis upon infection and to evaluate the consequences for osteoprogenitor cells. The results indicate that high levels (144 ng/ml) of BMP2 can be produced in non-osteoprogenitor cells (A549 cell line) by this method and the resultant protein appears to be three times more biologically active than the recombinant protein. Surprisingly, similar levels of BMP2 expression could not be achieved after transduction with Ad5BMP2 of either human bone marrow stromal cells or the mouse bone marrow stromal cell line W20-17. However, human bone marrow stromal cells cultured with 1 microM dexamethasone for four days, or further stimulated to become osteoblast-like cells with 50 microg/ml ascorbic acid, produced high levels of BMP2 upon Ad5BMP2 infection as compared to the undifferentiated cells. The increased production of BMP2 in adenovirus transduced cells following exposure to 1 microM dexamethasone was reduced if the cells were not given 50 microg/ml ascorbic acid. When bone marrow stromal cells were allowed to become confluent in culture prior to differentiation, BMP2 production in response to Ad5BMP2 infection was lost entirely. Furthermore, the increase in BMP2 synthesis seen during differentiation was greatly decreased when Ad5BMP2 was administered prior to dexamethasone treatment. In short, the efficiency of adenovirus mediated expression of BMP2 in bone marrow stromal cells appears to be dependent on the differentiation state of these cells.
Olmsted
E A
EA
Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX 77030, USA.
Blum
J S
JS
Rill
D
D
Yotnda
P
P
Gugala
Z
Z
Lindsey
R W
RW
Davis
A R
AR
D000256:Adenoviridae / Q000235:genetics* / Q000472:pathogenicity; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism*; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000235:genetics*; D002454:Cell Differentiation / Q000187:drug effects; D002472:Cell Transformation, Viral; D003907:Dexamethasone / Q000494:pharmacology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins; D011995:Recombination, Genetic / Q000235:genetics; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D016212:Transforming Growth Factor beta; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism
C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; D003907:Dexamethasone; D001205:Ascorbic Acid