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

机译:环状ADP核糖是人骨髓间充质干细胞中细胞内Ca2 +振荡的新型调节剂

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摘要

Bone marrow mesenchymal stem cells (MSCs) are a promising cell source for regenerative medicine. However, the cellular biology of these cells is not fully understood. The present study characterizes the cyclic ADP-ribose (cADPR)-mediated Ca2+ signals in human MSCs and finds that externally applied cADPR can increase the frequency of spontaneous intracellular Ca2+ (Ca2+i) oscillations. The increase was abrogated by a specific cADPR antagonist or an inositol trisphosphate receptor (IP3R) inhibitor, but not by ryanodine. In addition, the cADPR-induced increase of Ca2+i oscillation frequency was prevented by inhibitors of nucleoside transporter or by inhibitors of the transient receptor potential cation melastatin-2 (TRPM2) channel. RT-PCR revealed mRNAs for the nucleoside transporters, concentrative nucleoside transporters 1/2 and equilibrative nucleoside transporters 1/3, IP3R1/2/3 and the TRPM2 channel, but not those for ryanodine receptors and CD38 in human MSCs. Knockdown of the TRPM2 channel by specific short interference RNA abolished the effect of cADPR on the Ca2+i oscillation frequency, and prevented the stimulation of proliferation by cADPR. Moreover, cADPR remarkably increased phosphorylated extracellular-signal-regulated kinases 1/2 (ERK1/2), but not Akt or p38 mitogen-activated protein kinase (MAPK). However, cADPR had no effect on adipogenesis or osteogenesis in human MSCs. Our results indicate that cADPR is a novel regulator of Ca2+i oscillations in human MSCs. It permeates the cell membrane through the nucleoside transporters and increases Ca2+ oscillation via activation of the TRPM2 channel, resulting in enhanced phosphorylation of ERK1/2 and, thereby, stimulation of human MSC proliferation. This study delineates an alternate signalling pathway of cADPR that is distinct from its well-established role of serving as a Ca2+ messenger for mobilizing the internal Ca2+ stores. Whether cADPR can be used clinically for stimulating marrow function in patients with marrow disorders remains to be further studied.
机译:骨髓间充质干细胞(MSC)是再生医学的有希望的细胞来源。但是,这些细胞的细胞生物学尚未完全了解。本研究表征了人MSC中环ADP-核糖(cADPR)介导的Ca 2 + 信号,并发现外用cADPR可以增加自发性细胞内Ca 2 + (Ca 2 + i)振荡。特定的cADPR拮抗剂或肌醇三磷酸受体(IP3R)抑制剂可消除这种增加,但雷诺丹则不能。此外,cADPR诱导的Ca 2 + 振荡频率的增加被核苷转运蛋白的抑制剂或瞬时受体电位阳离子melastatin-2(TRPM2)通道的抑制剂所阻止。 RT-PCR揭示了人类MSC中核苷转运蛋白1/2,浓缩核苷转运蛋白1/2和平衡核苷转运蛋白1/3,IP3R1 / 2/3和TRPM2通道的mRNA,但没有关于莱丹碱受体和CD38的mRNA。特异性短干扰RNA敲低TRPM2通道消除了cADPR对Ca 2 + i振荡频率的影响,并阻止了cADPR刺激增殖。此外,cADPR显着增加了磷酸化的细胞外信号调节激酶1/2(ERK1 / 2),而不是Akt或p38丝裂原活化蛋白激酶(MAPK)。但是,cADPR对人MSC中的脂肪形成或成骨没有影响。我们的结果表明,cADPR是人类MSC中Ca 2 + i振荡的新型调节剂。它通过核苷转运蛋白渗透细胞膜,并通过激活TRPM2通道增加Ca 2 + 振荡,从而增强ERK1 / 2的磷酸化,从而刺激人MSC的增殖。这项研究描述了cADPR的另一种信号传导途径,该途径不同于其已确立的充当Ca 2 + 信使来动员内部Ca 2 + 存储的作用。 cADPR是否可在临床上用于刺激骨髓疾病患者的骨髓功能仍有待进一步研究。

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