The osteogenic or adipogenic lineage commitment of human mesenchymal stem cells is determined by protein kinase C delta

BMC Cell Biology

Dongchul Kang¹  Hang Thi Thuy Bui¹  Hee-Yeon Cho¹  Sooho Lee² 

[1] Department of Biomedical Gerontology, Graduate School of Hallym University, Chuncheon 200-702, Gangwon-do, Republic of Korea;Ilsong Institute of Life Science, Hallym University, Anyang 431-060, Gyeonggi-do, Republic of Korea

关键词: AMPK;    PKCδ;    Signal transduction;    Osteogenic differentiation;    hBMSCs;   

Others  :  1088718
DOI  :  10.1186/s12860-014-0042-4

 received in 2014-07-10, accepted in 2014-11-05,  发布年份 2014

【 摘 要 】

Background

Mesenchymal stem cells (MSCs) have the potential to differentiate into specialized cell lineages such as osteoblasts and adipocytes in vitro. There exists a reciprocal relationship between osteogenic and adipogenic differentiation of MSCs that an osteogenic phenotype occurs at the expense of an adipogenic phenotype and vice versa, which in turn influence one another’s phenotype through negative feedback loops. Thus, it is important to understand what signaling molecules modulate the lineage commitment of MSCs. Protein kinase C (PKC) plays a central role in cellular signal transduction for mediating diverse biological functions, and dysregulation of PKC activity is involved in various metabolic diseases including cancer, diabetes, and heart disease. Although the role of individual PKC isoforms has been investigated in various fields, the potential role of PKC in bone metabolism is not completely understood. In this study, we investigated the potential role of PKCδ in osteogenic lineage commitment of human bone marrow-derived mesenchymal stem cells (hBMSCs).

Results

We observed that expression and phosphorylation of PKCδ were increased during osteogenic differentiation of hBMSCs. Pharmacological inhibition and genetic ablation of PKCδ in hBMSCs resulted in a significant attenuation of osteogenic differentiation as evidenced by reduced ALP activity and ECM mineralization, as well as down-regulation of the expression of osteoblast-specific genes. These effects were also accompanied by induction of adipogenic differentiation and up-regulation of the expression of adipocyte-specific genes involved in lipid synthesis in osteogenic induction of hBMSCs. Additionally, the activation of AMPK, which is a key cellular energy sensor, induced osteogenesis of hBMSCs. However, the inhibition of AMPK activity by compound C did not affect the activation of PKCδ at all, indicating that there is no direct correlation between AMPK and PKCδ in osteogenesis of hBMSCs.

Conclusions

These results suggest that PKCδ is a critical regulator for the balance between osteogenesis and adipogenesis of hBMSCs and thus has a potential novel therapeutic target for the treatment of metabolic bone diseases.

【 授权许可】

   
2014 Lee et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Chamberlain G, Fox J, Ashton B, Middleton J: Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells 2007, 25(11):2739-2749.
• [2]Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR: Multilineage potential of adult human mesenchymal stem cells. Science 1999, 284(5411):143-147.
• [3]Uccelli A, Moretta L, Pistoia V: Mesenchymal stem cells in health and disease. Nat Rev Immunol 2008, 8(9):726-736.
• [4]Giordano A, Galderisi U, Marino IR: From the laboratory bench to the patient's bedside: an update on clinical trials with mesenchymal stem cells. J Cell Physiol 2007, 211(1):27-35.
• [5]Long F: Building strong bones: molecular regulation of the osteoblast lineage. Nat Rev Mol Cell Biol 2012, 13(1):27-38.
• [6]Stein GS, Lian JB, Van Wijnen AJ, Stein JL, Montecino M, Javed A, Zaidi SK, Young DW, Choi JY, Pockwinse SM: Runx2 control of organization, assembly and activity of the regulatory machinery for skeletal gene expression. Oncogene 2004, 23(24):4315-4329.
• [7]Arvidson K, Abdallah BM, Applegate LA, Baldini N, Cenni E, Gomez-Barrena E, Granchi D, Kassem M, Konttinen YT, Mustafa K, Pioletti DP, Sillat T, Finne-Wistrand A: Bone regeneration and stem cells. J Cell Mol Med 2011, 15(4):718-746.
• [8]Papachroni KK, Karatzas DN, Papavassiliou KA, Basdra EK, Papavassiliou AG: Mechanotransduction in osteoblast regulation and bone disease. Trends Mol Med 2009, 15(5):208-216.
• [9]Griner EM, Kazanietz MG: Protein kinase C and other diacylglycerol effectors in cancer. Nat Rev Cancer 2007, 7(4):281-294.
• [10]Teicher BA: Protein kinase C as a therapeutic target. Clin Cancer Res 2006, 12(18):5336-5345.
• [11]Geraldes P, King GL: Activation of protein kinase C isoforms and its impact on diabetic complications. Circ Res 2010, 106(8):1319-1331.
• [12]Rosse C, Linch M, Kermorgant S, Cameron AJ, Boeckeler K, Parker PJ: PKC and the control of localized signal dynamics. Nat Rev Mol Cell Biol 2010, 11(2):103-112.
• [13]Tu X, Joeng KS, Nakayama KI, Nakayama K, Rajagopal J, Carroll TJ, McMahon AP, Long F: Noncanonical Wnt signaling through G protein-linked PKCdelta activation promotes bone formation. Dev Cell 2007, 12(1):113-127.
• [14]Nakura A, Higuchi C, Yoshida K, Yoshikawa H: PKCalpha suppresses osteoblastic differentiation. Bone 2011, 48(3):476-484.
• [15]Park KH, Han DI, Rhee YH, Jeong SJ, Kim SH, Park YG: Protein kinase C betaII and delta/theta play critical roles in bone morphogenic protein-4-stimulated osteoblastic differentiation of MC3T3-E1 cells. Biochem Biophys Res Commun 2010, 403(1):7-12.
• [16]Khor EC, Abel T, Tickner J, Chim SM, Wang C, Cheng T, Ng B, Ng PY, Teguh DA, Kenny J, Yang X, Chen H, Nakayama KI, Nakayama K, Pavlos N, Zheng MH, Xu J: Loss of protein kinase C-delta protects against LPS-induced osteolysis owing to an intrinsic defect in osteoclastic bone resorption. PLoS One 2013, 8(8):e70815.
• [17]Jeyabalan J, Shah M, Viollet B, Chenu C: AMP-activated protein kinase pathway and bone metabolism. J Endocrinol 2012, 212(3):277-290.
• [18]Bair AM, Thippegowda PB, Freichel M, Cheng N, Ye RD, Vogel SM, Yu Y, Flockerzi V, Malik AB, Tiruppathi C: Ca2+ entry via TRPC channels is necessary for thrombin-induced NF-kappaB activation in endothelial cells through AMP-activated protein kinase and protein kinase Cdelta. J Biol Chem 2009, 284(1):563-574.
• [19]Mehla R, Bivalkar-Mehla S, Zhang R, Handy I, Albrecht H, Giri S, Nagarkatti P, Nagarkatti M, Chauhan A: Bryostatin modulates latent HIV-1 infection via PKC and AMPK signaling but inhibits acute infection in a receptor independent manner. PLoS One 2010, 5(6):e11160.
• [20]Kutner RH, Zhang XY, Reiser J: Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors. Nat Protoc 2009, 4(4):495-505.
• [21]Soltoff SP: Rottlerin: an inappropriate and ineffective inhibitor of PKCdelta. Trends Pharmacol Sci 2007, 28(9):453-458.
• [22]Takada I, Kouzmenko AP, Kato S: Wnt and PPARgamma signaling in osteoblastogenesis and adipogenesis. Nat Rev Rheumatol 2009, 5(8):442-447.
• [23]Duque G: Bone and fat connection in aging bone. Curr Opin Rheumatol 2008, 20(4):429-434.
• [24]Harada S, Rodan GA: Control of osteoblast function and regulation of bone mass. Nature 2003, 423(6937):349-355.
• [25]Rosen CJ, Klibanski A: Bone, fat, and body composition: evolving concepts in the pathogenesis of osteoporosis. Am J Med 2009, 122(5):409-414.
• [26]Zhou Y, Wang D, Li F, Shi J, Song J: Different roles of protein kinase C-betaI and -delta in the regulation of adipocyte differentiation. Int J Biochem Cell Biol 2006, 38(12):2151-2163.
• [27]Schmitz-Peiffer C: The tail wagging the dog–regulation of lipid metabolism by protein kinase C. FEBS J 2013, 280(21):5371-5383.
• [28]Mochly-Rosen D, Das K, Grimes KV: Protein kinase C, an elusive therapeutic target? Nat Rev Drug Discov 2012, 11(12):937-957.
• [29]Liu J, Someren E, Mentink A, Licht R, Dechering K, van Blitterswijk C, de Boer J: The effect of PKC activation and inhibition on osteogenic differentiation of human mesenchymal stem cells. J Tissue Eng Regen Med 2010, 4(5):329-339.
• [30]Lampasso JD, Chen W, Marzec N: The expression profile of PKC isoforms during MC3T3-E1 differentiation. Int J Mol Med 2006, 17(6):1125-1131.
• [31]Sanders JL, Stern PH: Expression and phorbol ester-induced down-regulation of protein kinase C isozymes in osteoblasts. J Bone Miner Res 1996, 11(12):1862-1872.
• [32]Kim HJ, Kim JH, Bae SC, Choi JY, Ryoo HM: The protein kinase C pathway plays a central role in the fibroblast growth factor-stimulated expression and transactivation activity of Runx2. J Biol Chem 2003, 278(1):319-326.
• [33]Kim BG, Kim HJ, Park HJ, Kim YJ, Yoon WJ, Lee SJ, Ryoo HM, Cho JY: Runx2 phosphorylation induced by fibroblast growth factor-2/protein kinase C pathways. Proteomics 2006, 6(4):1166-1174.
• [34]Murakami M, Horowitz A, Tang S, Ware JA, Simons M: Protein kinase C (PKC) delta regulates PKCalpha activity in a Syndecan-4-dependent manner. J Biol Chem 2002, 277(23):20367-20371.
• [35]Shui C, Spelsberg TC, Riggs BL, Khosla S: Changes in Runx2/Cbfa1 expression and activity during osteoblastic differentiation of human bone marrow stromal cells. J Bone Miner Res 2003, 18(2):213-221.
• [36]Jonason JH, Xiao G, Zhang M, Xing L, Chen D: Post-translational regulation of Runx2 in Bone and Cartilage. J Dent Res 2009, 88(8):693-703.
• [37]Lima F, Niger C, Hebert C, Stains JP: Connexin43 potentiates osteoblast responsiveness to fibroblast growth factor 2 via a protein kinase C-delta/Runx2-dependent mechanism. Mol Biol Cell 2009, 20(11):2697-2708.
• [38]Pantovic A, Krstic A, Janjetovic K, Kocic J, Harhaji-Trajkovic L, Bugarski D, Trajkovic V: Coordinated time-dependent modulation of AMPK/Akt/mTOR signaling and autophagy controls osteogenic differentiation of human mesenchymal stem cells. Bone 2013, 52(1):524-531.
• [39]Kim EK, Lim S, Park JM, Seo JK, Kim JH, Kim KT, Ryu SH, Suh PG: Human mesenchymal stem cell differentiation to the osteogenic or adipogenic lineage is regulated by AMP-activated protein kinase. J Cell Physiol 2012, 227(4):1680-1687.
• [40]Rosen CJ, Bouxsein ML: Mechanisms of disease: is osteoporosis the obesity of bone? Nat Clin Pract Rheumatol 2006, 2(1):35-43.
• [41]Zhao LJ, Jiang H, Papasian CJ, Maulik D, Drees B, Hamilton J, Deng HW: Correlation of obesity and osteoporosis: effect of fat mass on the determination of osteoporosis. J Bone Miner Res 2008, 23(1):17-29.
• [42]Rossi F, McNagny M, Smith G, Frampton J, Graf T: Lineage commitment of transformed haematopoietic progenitors is determined by the level of PKC activity. EMBO J 1996, 15(8):1894-1901.
• [43]Chen H, Liu X, Cao J, Zhang L, Hu X, Wang J: Role of SIRT1 and AMPK in mesenchymal stem cells differentiation. Ageing Res Rev 2014, 13:55-64.