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Urokinase-type plasminogen activator receptor interaction with β1 integrin is required for platelet-derived growth factor-AB-induced human mesenchymal stem/stromal cell migration.

Chabot V, Dromard C, Rico A, Langonné A, Gaillard J, Guilloton F, Casteilla L, Sensebé L - Stem Cell Res Ther (2015)

Bottom Line: We demonstrated that uPAR activation and its association with β1-integrin are required for PDGF-AB-induced migration.This phenomenon takes place in MSC derived from bone marrow and from adipose tissue.We showed that PDGF-AB downstream signaling requires other effector molecules in MSC such as the uPA/uPAR system and β1 integrin signaling pathway known for their role in migration.

View Article: PubMed Central - PubMed

Affiliation: EFS Centre-Atlantique, BP 40661, 37 206, Tours, Cedex 3, France. valerie.chabot@efs.sante.fr.

ABSTRACT

Introduction: Mesenchymal stem cells (MSC) are well described for their role in tissue regeneration following injury. Migratory properties of endogenous or administrated MSC are critical for tissue repair processes. Platelet-derived growth factor (PDGF) is a chemotactic growth factor that elicits mesenchymal cell migration. However, it is yet to be elucidated if signaling pathways other than direct activation of PDGF receptor (PDGF-R) are involved in PDGF-induced cell migration.

Methods: Knocking down and co-immunoprecipitation approaches were used to evaluate urokinase-type plasminogen activator receptor (uPAR) requirement and its interactions with proteins involved in migration mechanisms, in human MSC induced to migrate under PDGF-AB effect.

Results: We demonstrated that uPAR activation and its association with β1-integrin are required for PDGF-AB-induced migration. This phenomenon takes place in MSC derived from bone marrow and from adipose tissue.

Conclusions: We showed that PDGF-AB downstream signaling requires other effector molecules in MSC such as the uPA/uPAR system and β1 integrin signaling pathway known for their role in migration. These findings provide new insights in molecular mechanisms of PDGF-AB-induced migration of human MSC that may be relevant to control MSC function and tissue remodeling after injury.

No MeSH data available.


Related in: MedlinePlus

Regulation of BM-MSC migration by inflammatory molecules. Scratch test assay was performed on BM-MSC isolated from three different donors and seeded in plates coated with type I dermal collagen, cellular fibronectin, or vitronectin or without ECM. Pictures of scratches were taken immediately (T = 0) or 22 hours after culture (T = 22 h) in serum-free control medium or medium supplemented with IL-1β, IL-8, TNFα, or PDGF-AB. a Quantification of BM-MSC migration. Results expressed as percentages of wound closure at T = 22 h compared with T = 0 for each condition. Data expressed as mean ± SEM of three independent experiments (one donor per experiment), each performed in triplicate. *P <0.05. b, c One experiment representative of the three experiments is shown. ECM extracellular matrix, IL interleukin, PDGF-AB platelet-derived growth factor AB, TNFα tumor necrosis factor alpha
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Fig1: Regulation of BM-MSC migration by inflammatory molecules. Scratch test assay was performed on BM-MSC isolated from three different donors and seeded in plates coated with type I dermal collagen, cellular fibronectin, or vitronectin or without ECM. Pictures of scratches were taken immediately (T = 0) or 22 hours after culture (T = 22 h) in serum-free control medium or medium supplemented with IL-1β, IL-8, TNFα, or PDGF-AB. a Quantification of BM-MSC migration. Results expressed as percentages of wound closure at T = 22 h compared with T = 0 for each condition. Data expressed as mean ± SEM of three independent experiments (one donor per experiment), each performed in triplicate. *P <0.05. b, c One experiment representative of the three experiments is shown. ECM extracellular matrix, IL interleukin, PDGF-AB platelet-derived growth factor AB, TNFα tumor necrosis factor alpha

Mentions: To investigate which inflammatory molecules play a key role in BM-MSC migration, we used an in vitro scratch test assay in which plates were coated or not with three different ECM proteins overexpressed in wounding tissues including vitronectin, fibronectin, and type I dermal collagen. In the presence or absence of ECM, nontreated BM-MSC displayed poor migratory capacities 22 hours after the scratch (Control; Fig. 1a, b). IL-1β, IL-8, and TNFα had no effect on BM-MSC migration in the absence of ECM or on vitronectin, compared with nontreated BM-MSC (Control; Fig. 1a). IL-1β did not affect BM-MSC migration in any other conditions tested while IL-8 stimulated BM-MSC migration on collagen I only (12 % enhancement) and TNFα increased BM-MSC migration on fibronectin (13 % enhancement) and collagen I (10 % enhancement). The effects of IL-8 and TNFα on BM-MSC migration were far less than those of PDGF-AB. PDGF-AB significantly enhanced the migration of BM-MSC; 18 % enhancement in absence of ECM, 22 % on vitronectin, 36 % on fibronectin, and greater effects were observed on type I dermal collagen: 43 % enhancement (Fig. 1a, c). As migratory stimulation by PDGF-AB was observed for the three time points we tested (14, 18, and 22 hours) (data not shown), all of the following migration experiments were conducted 22 hours after scratch. We controlled so that proliferation of BM-MSC was not increased under any condition used for the scratch test assay (Figure S2A in Additional file 3). The effect of PDGF treatment on stimulation of MSC migration was confirmed using another migration assay, in transwell dishes (Figure S2B in Additional file 3). The results showed that PDGF-AB treatment for 22 hours increased by sixfold the number of cells that migrated on the lower face of the filters. These results indicate that BM-MSC migrate in response to several molecules known to be released in damaged tissue and that PDGF-AB is one of the most potent, especially on collagen I.Fig. 1


Urokinase-type plasminogen activator receptor interaction with β1 integrin is required for platelet-derived growth factor-AB-induced human mesenchymal stem/stromal cell migration.

Chabot V, Dromard C, Rico A, Langonné A, Gaillard J, Guilloton F, Casteilla L, Sensebé L - Stem Cell Res Ther (2015)

Regulation of BM-MSC migration by inflammatory molecules. Scratch test assay was performed on BM-MSC isolated from three different donors and seeded in plates coated with type I dermal collagen, cellular fibronectin, or vitronectin or without ECM. Pictures of scratches were taken immediately (T = 0) or 22 hours after culture (T = 22 h) in serum-free control medium or medium supplemented with IL-1β, IL-8, TNFα, or PDGF-AB. a Quantification of BM-MSC migration. Results expressed as percentages of wound closure at T = 22 h compared with T = 0 for each condition. Data expressed as mean ± SEM of three independent experiments (one donor per experiment), each performed in triplicate. *P <0.05. b, c One experiment representative of the three experiments is shown. ECM extracellular matrix, IL interleukin, PDGF-AB platelet-derived growth factor AB, TNFα tumor necrosis factor alpha
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4588680&req=5

Fig1: Regulation of BM-MSC migration by inflammatory molecules. Scratch test assay was performed on BM-MSC isolated from three different donors and seeded in plates coated with type I dermal collagen, cellular fibronectin, or vitronectin or without ECM. Pictures of scratches were taken immediately (T = 0) or 22 hours after culture (T = 22 h) in serum-free control medium or medium supplemented with IL-1β, IL-8, TNFα, or PDGF-AB. a Quantification of BM-MSC migration. Results expressed as percentages of wound closure at T = 22 h compared with T = 0 for each condition. Data expressed as mean ± SEM of three independent experiments (one donor per experiment), each performed in triplicate. *P <0.05. b, c One experiment representative of the three experiments is shown. ECM extracellular matrix, IL interleukin, PDGF-AB platelet-derived growth factor AB, TNFα tumor necrosis factor alpha
Mentions: To investigate which inflammatory molecules play a key role in BM-MSC migration, we used an in vitro scratch test assay in which plates were coated or not with three different ECM proteins overexpressed in wounding tissues including vitronectin, fibronectin, and type I dermal collagen. In the presence or absence of ECM, nontreated BM-MSC displayed poor migratory capacities 22 hours after the scratch (Control; Fig. 1a, b). IL-1β, IL-8, and TNFα had no effect on BM-MSC migration in the absence of ECM or on vitronectin, compared with nontreated BM-MSC (Control; Fig. 1a). IL-1β did not affect BM-MSC migration in any other conditions tested while IL-8 stimulated BM-MSC migration on collagen I only (12 % enhancement) and TNFα increased BM-MSC migration on fibronectin (13 % enhancement) and collagen I (10 % enhancement). The effects of IL-8 and TNFα on BM-MSC migration were far less than those of PDGF-AB. PDGF-AB significantly enhanced the migration of BM-MSC; 18 % enhancement in absence of ECM, 22 % on vitronectin, 36 % on fibronectin, and greater effects were observed on type I dermal collagen: 43 % enhancement (Fig. 1a, c). As migratory stimulation by PDGF-AB was observed for the three time points we tested (14, 18, and 22 hours) (data not shown), all of the following migration experiments were conducted 22 hours after scratch. We controlled so that proliferation of BM-MSC was not increased under any condition used for the scratch test assay (Figure S2A in Additional file 3). The effect of PDGF treatment on stimulation of MSC migration was confirmed using another migration assay, in transwell dishes (Figure S2B in Additional file 3). The results showed that PDGF-AB treatment for 22 hours increased by sixfold the number of cells that migrated on the lower face of the filters. These results indicate that BM-MSC migrate in response to several molecules known to be released in damaged tissue and that PDGF-AB is one of the most potent, especially on collagen I.Fig. 1

Bottom Line: We demonstrated that uPAR activation and its association with β1-integrin are required for PDGF-AB-induced migration.This phenomenon takes place in MSC derived from bone marrow and from adipose tissue.We showed that PDGF-AB downstream signaling requires other effector molecules in MSC such as the uPA/uPAR system and β1 integrin signaling pathway known for their role in migration.

View Article: PubMed Central - PubMed

Affiliation: EFS Centre-Atlantique, BP 40661, 37 206, Tours, Cedex 3, France. valerie.chabot@efs.sante.fr.

ABSTRACT

Introduction: Mesenchymal stem cells (MSC) are well described for their role in tissue regeneration following injury. Migratory properties of endogenous or administrated MSC are critical for tissue repair processes. Platelet-derived growth factor (PDGF) is a chemotactic growth factor that elicits mesenchymal cell migration. However, it is yet to be elucidated if signaling pathways other than direct activation of PDGF receptor (PDGF-R) are involved in PDGF-induced cell migration.

Methods: Knocking down and co-immunoprecipitation approaches were used to evaluate urokinase-type plasminogen activator receptor (uPAR) requirement and its interactions with proteins involved in migration mechanisms, in human MSC induced to migrate under PDGF-AB effect.

Results: We demonstrated that uPAR activation and its association with β1-integrin are required for PDGF-AB-induced migration. This phenomenon takes place in MSC derived from bone marrow and from adipose tissue.

Conclusions: We showed that PDGF-AB downstream signaling requires other effector molecules in MSC such as the uPA/uPAR system and β1 integrin signaling pathway known for their role in migration. These findings provide new insights in molecular mechanisms of PDGF-AB-induced migration of human MSC that may be relevant to control MSC function and tissue remodeling after injury.

No MeSH data available.


Related in: MedlinePlus