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Globular adiponectin as a complete mesoangioblast regulator: role in proliferation, survival, motility, and skeletal muscle differentiation.

Fiaschi T, Tedesco FS, Giannoni E, Diaz-Manera J, Parri M, Cossu G, Chiarugi P - Mol. Biol. Cell (2010)

Bottom Line: Mesoangioblasts are progenitor endowed with multipotent mesoderm differentiation ability.The adipokine drives mesoangioblasts to entry cell cycle and strongly counteracts the apoptotic process triggered by growth factor withdrawal, thereby serving as an activating and prosurvival stem cell factor.We conclude that adiponectin exerts several advantageous effects on mesoangioblasts, potentially valuable to improve their efficacy in cell based therapies of diseased muscles.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Science, University of Florence, 50134 Florence, Italy.

ABSTRACT
Mesoangioblasts are progenitor endowed with multipotent mesoderm differentiation ability. Despite the promising results obtained with mesoangioblast transplantation in muscle dystrophy, an improvement of their efficient engrafting and survival within damaged muscles, as well as their ex vivo activation/expansion and commitment toward myogenic lineage, is highly needed and should greatly increase their therapeutic potential. We show that globular adiponectin, an adipokine endowed with metabolic and differentiating functions for muscles, regulates vital cues of mesoangioblast cell biology. The adipokine drives mesoangioblasts to entry cell cycle and strongly counteracts the apoptotic process triggered by growth factor withdrawal, thereby serving as an activating and prosurvival stem cell factor. In addition, adiponectin provides a specific protection against anoikis, the apoptotic death due to lack of anchorage to extracellular matrix, suggesting a key protective role for these nonresident stem cells after systemic injection. Finally, adiponectin behaves as a chemoattractive factor toward mature myotubes and stimulates their differentiation toward the skeletal muscle lineage, serving as a positive regulator in mesoangioblast homing to injured or diseased muscles. We conclude that adiponectin exerts several advantageous effects on mesoangioblasts, potentially valuable to improve their efficacy in cell based therapies of diseased muscles.

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Adiponectin induces the growth of mesoangioblasts. (A) Mesoangioblasts were serum-deprived for 24 h, and where indicated, adiponectin (1 μg/ml) was added to serum-free medium for 72 h. Cells were then counted using an hemocytometer. (B) Analysis of [3H]thymidine incorporation by mesoangioblasts after the treatment with adiponectin (1 μg/ml). Cells were treated as in A, and [3H]thymidine was added during the last 2 h of incubation. These results correspond to the mean of four different experiments. *p < 0.001 and **p < 0.005 versus control. (C–E) Analysis of the signaling pathways activated by adiponectin stimulation. Mesoangioblasts were serum-deprived overnight and then stimulated with adiponectin (1 μg/ml) for the indicated period. Immunoblot analysis for the detection of the phosphorylation level of p42/p44 MAPK (Thr202/Tyr204), Akt (Ser473), p38 MAPK (Thr180/Tyr182), and AMPK (Thr182) was performed using specific phospho-antibodies. p42/p44 MAPK (C), p38 (D), and AMPK (E) immunoblots were used for normalization. Bar graph represents the phosphorylation level of the signaling proteins calculated by the ratio between the phosphorylated and total protein obtained in four different experiments. *p < 0.001 and **p < 0.005 versus time 0.
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Figure 2: Adiponectin induces the growth of mesoangioblasts. (A) Mesoangioblasts were serum-deprived for 24 h, and where indicated, adiponectin (1 μg/ml) was added to serum-free medium for 72 h. Cells were then counted using an hemocytometer. (B) Analysis of [3H]thymidine incorporation by mesoangioblasts after the treatment with adiponectin (1 μg/ml). Cells were treated as in A, and [3H]thymidine was added during the last 2 h of incubation. These results correspond to the mean of four different experiments. *p < 0.001 and **p < 0.005 versus control. (C–E) Analysis of the signaling pathways activated by adiponectin stimulation. Mesoangioblasts were serum-deprived overnight and then stimulated with adiponectin (1 μg/ml) for the indicated period. Immunoblot analysis for the detection of the phosphorylation level of p42/p44 MAPK (Thr202/Tyr204), Akt (Ser473), p38 MAPK (Thr180/Tyr182), and AMPK (Thr182) was performed using specific phospho-antibodies. p42/p44 MAPK (C), p38 (D), and AMPK (E) immunoblots were used for normalization. Bar graph represents the phosphorylation level of the signaling proteins calculated by the ratio between the phosphorylated and total protein obtained in four different experiments. *p < 0.001 and **p < 0.005 versus time 0.

Mentions: To test whether the hormone can act as a growth factor (GF) in mesoangioblasts, serum-deprived cells were cultured for 72 h with 1 μg/ml adiponectin and then counted with a hemocytometer. We observed that the treatment of mesoangioblasts with adiponectin leads to a 100% increase in cell number (Figure 2A). The effect of adiponectin on mesoangioblast proliferation was then evaluated by [3H]thymidine incorporation. Our results showed that adiponectin treatment induces ∼50% of increase in thymidine incorporation (Figure 2B), thereby demonstrating that adiponectin acts as GF in mesoangioblasts. In keeping with these observations, adiponectin is able to elicit a sustained activation of the p42/p44 MAPK two master regulators of mitogenic signaling (Figure 2C).


Globular adiponectin as a complete mesoangioblast regulator: role in proliferation, survival, motility, and skeletal muscle differentiation.

Fiaschi T, Tedesco FS, Giannoni E, Diaz-Manera J, Parri M, Cossu G, Chiarugi P - Mol. Biol. Cell (2010)

Adiponectin induces the growth of mesoangioblasts. (A) Mesoangioblasts were serum-deprived for 24 h, and where indicated, adiponectin (1 μg/ml) was added to serum-free medium for 72 h. Cells were then counted using an hemocytometer. (B) Analysis of [3H]thymidine incorporation by mesoangioblasts after the treatment with adiponectin (1 μg/ml). Cells were treated as in A, and [3H]thymidine was added during the last 2 h of incubation. These results correspond to the mean of four different experiments. *p < 0.001 and **p < 0.005 versus control. (C–E) Analysis of the signaling pathways activated by adiponectin stimulation. Mesoangioblasts were serum-deprived overnight and then stimulated with adiponectin (1 μg/ml) for the indicated period. Immunoblot analysis for the detection of the phosphorylation level of p42/p44 MAPK (Thr202/Tyr204), Akt (Ser473), p38 MAPK (Thr180/Tyr182), and AMPK (Thr182) was performed using specific phospho-antibodies. p42/p44 MAPK (C), p38 (D), and AMPK (E) immunoblots were used for normalization. Bar graph represents the phosphorylation level of the signaling proteins calculated by the ratio between the phosphorylated and total protein obtained in four different experiments. *p < 0.001 and **p < 0.005 versus time 0.
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Related In: Results  -  Collection

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Figure 2: Adiponectin induces the growth of mesoangioblasts. (A) Mesoangioblasts were serum-deprived for 24 h, and where indicated, adiponectin (1 μg/ml) was added to serum-free medium for 72 h. Cells were then counted using an hemocytometer. (B) Analysis of [3H]thymidine incorporation by mesoangioblasts after the treatment with adiponectin (1 μg/ml). Cells were treated as in A, and [3H]thymidine was added during the last 2 h of incubation. These results correspond to the mean of four different experiments. *p < 0.001 and **p < 0.005 versus control. (C–E) Analysis of the signaling pathways activated by adiponectin stimulation. Mesoangioblasts were serum-deprived overnight and then stimulated with adiponectin (1 μg/ml) for the indicated period. Immunoblot analysis for the detection of the phosphorylation level of p42/p44 MAPK (Thr202/Tyr204), Akt (Ser473), p38 MAPK (Thr180/Tyr182), and AMPK (Thr182) was performed using specific phospho-antibodies. p42/p44 MAPK (C), p38 (D), and AMPK (E) immunoblots were used for normalization. Bar graph represents the phosphorylation level of the signaling proteins calculated by the ratio between the phosphorylated and total protein obtained in four different experiments. *p < 0.001 and **p < 0.005 versus time 0.
Mentions: To test whether the hormone can act as a growth factor (GF) in mesoangioblasts, serum-deprived cells were cultured for 72 h with 1 μg/ml adiponectin and then counted with a hemocytometer. We observed that the treatment of mesoangioblasts with adiponectin leads to a 100% increase in cell number (Figure 2A). The effect of adiponectin on mesoangioblast proliferation was then evaluated by [3H]thymidine incorporation. Our results showed that adiponectin treatment induces ∼50% of increase in thymidine incorporation (Figure 2B), thereby demonstrating that adiponectin acts as GF in mesoangioblasts. In keeping with these observations, adiponectin is able to elicit a sustained activation of the p42/p44 MAPK two master regulators of mitogenic signaling (Figure 2C).

Bottom Line: Mesoangioblasts are progenitor endowed with multipotent mesoderm differentiation ability.The adipokine drives mesoangioblasts to entry cell cycle and strongly counteracts the apoptotic process triggered by growth factor withdrawal, thereby serving as an activating and prosurvival stem cell factor.We conclude that adiponectin exerts several advantageous effects on mesoangioblasts, potentially valuable to improve their efficacy in cell based therapies of diseased muscles.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Science, University of Florence, 50134 Florence, Italy.

ABSTRACT
Mesoangioblasts are progenitor endowed with multipotent mesoderm differentiation ability. Despite the promising results obtained with mesoangioblast transplantation in muscle dystrophy, an improvement of their efficient engrafting and survival within damaged muscles, as well as their ex vivo activation/expansion and commitment toward myogenic lineage, is highly needed and should greatly increase their therapeutic potential. We show that globular adiponectin, an adipokine endowed with metabolic and differentiating functions for muscles, regulates vital cues of mesoangioblast cell biology. The adipokine drives mesoangioblasts to entry cell cycle and strongly counteracts the apoptotic process triggered by growth factor withdrawal, thereby serving as an activating and prosurvival stem cell factor. In addition, adiponectin provides a specific protection against anoikis, the apoptotic death due to lack of anchorage to extracellular matrix, suggesting a key protective role for these nonresident stem cells after systemic injection. Finally, adiponectin behaves as a chemoattractive factor toward mature myotubes and stimulates their differentiation toward the skeletal muscle lineage, serving as a positive regulator in mesoangioblast homing to injured or diseased muscles. We conclude that adiponectin exerts several advantageous effects on mesoangioblasts, potentially valuable to improve their efficacy in cell based therapies of diseased muscles.

Show MeSH
Related in: MedlinePlus