TY - JOUR
T1 - Prediction of sphingosine 1-phosphate-stimulated endothelial cell migraton rates using biochemical measurements
AU - Alford, Shannon K.
AU - Wang, Yumei
AU - Feng, Yunfeng
AU - Longmore, Gregory D.
AU - Elbert, Donald L.
N1 - Funding Information:
We gratefully acknowledge funding from NIH, HL085364 (DLE), CA085839 (GDL), and GM080673 (GDL). We thank Sheila Stewart for technical guidance with hTERT tranformations, and Doug Lauf-fenburger for use of microscopy equipment.
PY - 2010/8
Y1 - 2010/8
N2 - The ability to predict endothelial cell migration rates may aid in the design of biomaterials that endothelialize following implantation. However, the complexity of the signaling response to migration-promoting stimuli such as sphingosine 1-phosphate (S1P) makes such predictions quite challenging. A number of signaling pathways impact S1Pmediated cell migration, including the Akt and Src pathways, which both affect activation of the small GTPase Rac. Rac activation promotes the formation of lamellipodia, and thus should be intimately linked to cell migration rates. In immortalized endothelial cells, expression of proteins that inhibit Akt, Src, and Rac (PTEN, CSK, and β2-chimaerin, respectively) was decreased usingRNAinterference, resulting in increases in the basal level of activation of Akt, Src, and Rac. Cells were scrape-wounded and stimulated with 1 μM S1P. The timecourse of Akt, Src, and Rac activation was followed over 2 h in the perturbed cells, while migration into the scrape wound was measured over 6 h. Rac activation at 120 min post-stimulation was highly correlated with the mean migration rate of cells, but only in cells stimulated with S1P. Using partial least squares regression, the migration rate of cells into the scrapewound was found to be highly correlated with the magnitude of the early Akt peak (e.g., 5-15 min post-stimulation). These results demonstrated that biochemical measurements might be useful in predicting rates of endothelial cell migration.
AB - The ability to predict endothelial cell migration rates may aid in the design of biomaterials that endothelialize following implantation. However, the complexity of the signaling response to migration-promoting stimuli such as sphingosine 1-phosphate (S1P) makes such predictions quite challenging. A number of signaling pathways impact S1Pmediated cell migration, including the Akt and Src pathways, which both affect activation of the small GTPase Rac. Rac activation promotes the formation of lamellipodia, and thus should be intimately linked to cell migration rates. In immortalized endothelial cells, expression of proteins that inhibit Akt, Src, and Rac (PTEN, CSK, and β2-chimaerin, respectively) was decreased usingRNAinterference, resulting in increases in the basal level of activation of Akt, Src, and Rac. Cells were scrape-wounded and stimulated with 1 μM S1P. The timecourse of Akt, Src, and Rac activation was followed over 2 h in the perturbed cells, while migration into the scrape wound was measured over 6 h. Rac activation at 120 min post-stimulation was highly correlated with the mean migration rate of cells, but only in cells stimulated with S1P. Using partial least squares regression, the migration rate of cells into the scrapewound was found to be highly correlated with the magnitude of the early Akt peak (e.g., 5-15 min post-stimulation). These results demonstrated that biochemical measurements might be useful in predicting rates of endothelial cell migration.
KW - Biochemistry
KW - Endothelial cell
KW - Migration
KW - Modeling
KW - Partial least squares regression
UR - http://www.scopus.com/inward/record.url?scp=77955172214&partnerID=8YFLogxK
U2 - 10.1007/s10439-010-0014-6
DO - 10.1007/s10439-010-0014-6
M3 - Article
C2 - 20358290
AN - SCOPUS:77955172214
SN - 0090-6964
VL - 38
SP - 2775
EP - 2790
JO - Annals of biomedical engineering
JF - Annals of biomedical engineering
IS - 8
ER -