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Cyclic ADP ribose is a novel regulator of intracellular Ca2+ oscillations in human bone marrow mesenchymal stem cells.

Tao R, Sun HY, Lau CP, Tse HF, Lee HC, Li GR - J. Cell. Mol. Med. (2011)

Bottom Line: However, cADPR had no effect on adipogenesis or osteogenesis in human MSCs.Our results indicate that cADPR is a novel regulator of Ca(2+) (i) oscillations in human MSCs.It permeates the cell membrane through the nucleoside transporters and increases Ca(2+) oscillation via activation of the TRPM2 channel, resulting in enhanced phosphorylation of ERK1/2 and, thereby, stimulation of human MSC proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.

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Cyclic ADP ribose and proliferation-related kinases. (A) ERK1/2 phosphorylation at Thr185/Tyr187 was enhanced in the presence of cADPR (10 and 50 μM) for 60 min. and 8-Br-cADPR (100 μM) reduced the effect (upper panels). Mean values (lower panel) for ratio of p-ERK1/2:total ERK1/2 (n = 3, *P < 0.05, **P < 0.01 versus 0 μM cADPR). (B) Time-dependent effect of cADPR on ERK1/2 phosphorylation and mean values for ratio of p-ERK1/2:total ERK1/2 in the presence of cADPR for 30, 60, 120 and 180 min. (n = 3, **P < 0.01 versus 0 μM cADPR). (C) ERK1/2 phosphorylation and mean values for ratio of p-ERK1/2:total ERK1/2 in the presence of cADPR in the cells transfected with control siRNA or TRPM2 siRNA for 72 hrs (n = 3, **P < 0.01 versus 0 μM cADPR). (D) Akt1 phosphorylation at Thr308 and Ser473 was not affected by cADPR (10 and 50 μM for 60 min.). (E) Mean values for ratio of p-Akt(308):total Akt (n = 3) and p-Akt(473):total Akt (n = 3). (F) p38 MAPK phosphorylation levels at Thr180/Tyr182 were not changed when incubated with cADPR (10 and 50 μM) for 60 min. (n = 3). (G) JNK phosphorylation levels at Thr183/Tyr185 were not changed when incubated with cADPR (10 and 50 μM) for 60 min. (n = 3).
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fig07: Cyclic ADP ribose and proliferation-related kinases. (A) ERK1/2 phosphorylation at Thr185/Tyr187 was enhanced in the presence of cADPR (10 and 50 μM) for 60 min. and 8-Br-cADPR (100 μM) reduced the effect (upper panels). Mean values (lower panel) for ratio of p-ERK1/2:total ERK1/2 (n = 3, *P < 0.05, **P < 0.01 versus 0 μM cADPR). (B) Time-dependent effect of cADPR on ERK1/2 phosphorylation and mean values for ratio of p-ERK1/2:total ERK1/2 in the presence of cADPR for 30, 60, 120 and 180 min. (n = 3, **P < 0.01 versus 0 μM cADPR). (C) ERK1/2 phosphorylation and mean values for ratio of p-ERK1/2:total ERK1/2 in the presence of cADPR in the cells transfected with control siRNA or TRPM2 siRNA for 72 hrs (n = 3, **P < 0.01 versus 0 μM cADPR). (D) Akt1 phosphorylation at Thr308 and Ser473 was not affected by cADPR (10 and 50 μM for 60 min.). (E) Mean values for ratio of p-Akt(308):total Akt (n = 3) and p-Akt(473):total Akt (n = 3). (F) p38 MAPK phosphorylation levels at Thr180/Tyr182 were not changed when incubated with cADPR (10 and 50 μM) for 60 min. (n = 3). (G) JNK phosphorylation levels at Thr183/Tyr185 were not changed when incubated with cADPR (10 and 50 μM) for 60 min. (n = 3).

Mentions: To determine whether the cADPR-induced increase of proliferation is mediated by MAP kinases, we investigated the phosphorylated levels of the ERK1/2 and p38 MAP, and survival kinase Akt. Figure 7A shows that a 30-min incubation with cADPR (10 and 50 μM) increased the phosphorylation level of ERK1/2 (Thr185/Tyr187) in human MSCs, and the effect was countered by 8-Br-cADPR (Fig. 7A). The effect of cADPR on phosphorylated ERK1/2 was time dependent. The increase of ERK1/2 phosphrylation level was maximal at 60 min., and the effect remained significant at 180 min. (Fig. 7B). However, the increase of phosphorylated ERK1/2 by cADPR was remarkably reduced in the cells transfected with molecule A of TRPM2 siRNAs. Cyclic ADPR (50 μM, 60 min. incubation) only induced a slight increase of phosphorylated level of ERK1/2 in TRPM2 siRNA (n = 3, P = NS) (Fig. 7C).


Cyclic ADP ribose is a novel regulator of intracellular Ca2+ oscillations in human bone marrow mesenchymal stem cells.

Tao R, Sun HY, Lau CP, Tse HF, Lee HC, Li GR - J. Cell. Mol. Med. (2011)

Cyclic ADP ribose and proliferation-related kinases. (A) ERK1/2 phosphorylation at Thr185/Tyr187 was enhanced in the presence of cADPR (10 and 50 μM) for 60 min. and 8-Br-cADPR (100 μM) reduced the effect (upper panels). Mean values (lower panel) for ratio of p-ERK1/2:total ERK1/2 (n = 3, *P < 0.05, **P < 0.01 versus 0 μM cADPR). (B) Time-dependent effect of cADPR on ERK1/2 phosphorylation and mean values for ratio of p-ERK1/2:total ERK1/2 in the presence of cADPR for 30, 60, 120 and 180 min. (n = 3, **P < 0.01 versus 0 μM cADPR). (C) ERK1/2 phosphorylation and mean values for ratio of p-ERK1/2:total ERK1/2 in the presence of cADPR in the cells transfected with control siRNA or TRPM2 siRNA for 72 hrs (n = 3, **P < 0.01 versus 0 μM cADPR). (D) Akt1 phosphorylation at Thr308 and Ser473 was not affected by cADPR (10 and 50 μM for 60 min.). (E) Mean values for ratio of p-Akt(308):total Akt (n = 3) and p-Akt(473):total Akt (n = 3). (F) p38 MAPK phosphorylation levels at Thr180/Tyr182 were not changed when incubated with cADPR (10 and 50 μM) for 60 min. (n = 3). (G) JNK phosphorylation levels at Thr183/Tyr185 were not changed when incubated with cADPR (10 and 50 μM) for 60 min. (n = 3).
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fig07: Cyclic ADP ribose and proliferation-related kinases. (A) ERK1/2 phosphorylation at Thr185/Tyr187 was enhanced in the presence of cADPR (10 and 50 μM) for 60 min. and 8-Br-cADPR (100 μM) reduced the effect (upper panels). Mean values (lower panel) for ratio of p-ERK1/2:total ERK1/2 (n = 3, *P < 0.05, **P < 0.01 versus 0 μM cADPR). (B) Time-dependent effect of cADPR on ERK1/2 phosphorylation and mean values for ratio of p-ERK1/2:total ERK1/2 in the presence of cADPR for 30, 60, 120 and 180 min. (n = 3, **P < 0.01 versus 0 μM cADPR). (C) ERK1/2 phosphorylation and mean values for ratio of p-ERK1/2:total ERK1/2 in the presence of cADPR in the cells transfected with control siRNA or TRPM2 siRNA for 72 hrs (n = 3, **P < 0.01 versus 0 μM cADPR). (D) Akt1 phosphorylation at Thr308 and Ser473 was not affected by cADPR (10 and 50 μM for 60 min.). (E) Mean values for ratio of p-Akt(308):total Akt (n = 3) and p-Akt(473):total Akt (n = 3). (F) p38 MAPK phosphorylation levels at Thr180/Tyr182 were not changed when incubated with cADPR (10 and 50 μM) for 60 min. (n = 3). (G) JNK phosphorylation levels at Thr183/Tyr185 were not changed when incubated with cADPR (10 and 50 μM) for 60 min. (n = 3).
Mentions: To determine whether the cADPR-induced increase of proliferation is mediated by MAP kinases, we investigated the phosphorylated levels of the ERK1/2 and p38 MAP, and survival kinase Akt. Figure 7A shows that a 30-min incubation with cADPR (10 and 50 μM) increased the phosphorylation level of ERK1/2 (Thr185/Tyr187) in human MSCs, and the effect was countered by 8-Br-cADPR (Fig. 7A). The effect of cADPR on phosphorylated ERK1/2 was time dependent. The increase of ERK1/2 phosphrylation level was maximal at 60 min., and the effect remained significant at 180 min. (Fig. 7B). However, the increase of phosphorylated ERK1/2 by cADPR was remarkably reduced in the cells transfected with molecule A of TRPM2 siRNAs. Cyclic ADPR (50 μM, 60 min. incubation) only induced a slight increase of phosphorylated level of ERK1/2 in TRPM2 siRNA (n = 3, P = NS) (Fig. 7C).

Bottom Line: However, cADPR had no effect on adipogenesis or osteogenesis in human MSCs.Our results indicate that cADPR is a novel regulator of Ca(2+) (i) oscillations in human MSCs.It permeates the cell membrane through the nucleoside transporters and increases Ca(2+) oscillation via activation of the TRPM2 channel, resulting in enhanced phosphorylation of ERK1/2 and, thereby, stimulation of human MSC proliferation.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.

Show MeSH