Restricted accessResearch articleFirst published online 2013-02
Activated NF-κB in Bone Marrow Mesenchymal Stem Cells from Systemic Lupus Erythematosus Patients Inhibits Osteogenic Differentiation Through Downregulating Smad Signaling
Osteoporosis in patients with systemic lupus erythematosus (SLE) is thought to be the result of accelerated osteoclastogenesis induced by pro-inflammatory cytokines such as tumor necrosis factor (TNF). However, the molecular mechanisms involved in the osteoblastogenesis in SLE patients are not fully understood. We investigated the bone morphogenetic protein-2 (BMP-2)-induced osteoblastic capacity of bone marrow-derived mesenchymal stem cells (BMMSCs) from SLE patients and the TNF signaling system in determining BMP-2-induced regulatory pathways. It showed that the capacity of osteogenic differentiation of BMMSCs from SLE patients was reduced compared with that from healthy controls. The nuclear factor κB (NF-κB) signaling was activated while the BMP/Smad pathway was repressed in BMMSCs from SLE patients. TNF activated NF-κB pathway and inhibited the phosphorylation of Smad 1/5/8 and BMP-2-induced osteoblastic differentiation in BMMSCs from normal controls, while addition of pyrollidine dithiocarbamate (PDTC), an NF-κB inhibitor, to SLE-BMMSCs could partially reverse these effects. Thus, our findings have shown that the activated NF-κB pathway in SLE-BMMSCs inhibits the BMP-2-induced osteoblastic differentiation through BMP/Smad signaling pathway, suggesting that the impaired osteoblastic differentiation may participate in the pathology of osteoporosis in SLE patients.
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References
1.
SinigagliaL, VarennaM, BinelliL, ZucchiF, GhiringhellaD, GallazziM, LimontaM, ZeniS, FantiniF. 1999. Determinants of bone mass in systemic lupus erythematosus: a cross sectional study on premenopausal women. J Rheumatol, 26:1280–1284.
2.
KallaAA, FataarAB, JessopSJ, BewerungeL. 1993. Loss of trabecular bone mineral density in systemic lupus erythematosus. Arthritis Rheum, 36:1726–1734.
3.
PetriM. 1995. Musculoskeletal complications of systemic lupus erythematosus in the Hopkins Lupus Cohort: an update. Arthritis Care Res, 8:137–145.
4.
FeldmannM, BrennanFM, MainiRN. 1996. Role of cytokines in rheumatoid arthritis. Annu Rev Immunol, 14:397–440.
5.
Studnicka-BenkeA, SteinerG, PeteraP, SmolenJS. 1996. Tumour necrosis factor alpha and its soluble receptors parallel clinical disease and autoimmune activity in systemic lupus erythematosus. Br J Rheumatol, 35:1067–1074.
6.
AringerM, SmolenJS. 2003. SLE—complex cytokine effects in a complex autoimmune disease: tumor necrosis factor in systemic lupus erythematosus. Arthritis Res Ther, 5:172–177.
7.
LiP, SchwarzEM, O'KeefeRJ, MaL, BoyceBF, XingL. 2004. RANK signaling is not required for TNFalpha-mediated increase in CD11(hi) osteoclast precursors but is essential for mature osteoclast formation in TNFalpha-mediated inflammatory arthritis. J Bone Miner Res, 19:207–213.
8.
TakayanagiH, OgasawaraK, HidaS, ChibaT, MurataS, SatoK, TakaokaA, YokochiT, OdaHet al.2000. T-cell-mediated regulation of osteoclastogenesis by signalling cross-talk between RANKL and IFN-gamma. Nature, 408:600–605.
9.
TheillLE, BoyleWJ, PenningerJM. 2002. RANK-L and RANK: T cells, bone loss, and mammalian evolution. Annu Rev Immunol, 20:795–823.
10.
SudaT, TakahashiN, UdagawaN, JimiE, GillespieMT, MartinTJ. 1999. Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr Rev, 20:345–357.
11.
RodanGA, MartinTJ. 1982. Role of osteoblasts in hormonal control of bone resorption—a hypothesis. Calcif Tissue Int, 34:311.
12.
GilbertLC, RubinJ, NanesMS. 2005. The p55 TNF receptor mediates TNF inhibition of osteoblast differentiation independently of apoptosis. Am J Physiol Endocrinol Metab, 288:E1011–E1018.
13.
HessK, UshmorovA, FiedlerJ, BrennerRE, WirthT. 2009. TNFalpha promotes osteogenic differentiation of human mesenchymal stem cells by triggering the NF-kappaB signaling pathway. Bone, 45:367–376.
14.
ChoHH, ShinKK, KimYJ, SongJS, KimJM, BaeYC, KimCD, JungJS. 2010. NF-kappaB activation stimulates osteogenic differentiation of mesenchymal stem cells derived from human adipose tissue by increasing TAZ expression. J Cell Physiol, 223:168–177.
15.
GilbertL, HeX, FarmerP, BodenS, KozlowskiM, RubinJ, NanesMS. 2000. Inhibition of osteoblast differentiation by tumor necrosis factor-alpha. Endocrinology, 141:3956–3964.
16.
NanesMS. 2003. Tumor necrosis factor-alpha: molecular and cellular mechanisms in skeletal pathology. Gene, 321:1–15.
17.
NakaseT, TakaokaK, MasuharaK, ShimizuK, YoshikawaH, OchiT. 1997. Interleukin-1 beta enhances and tumor necrosis factor-alpha inhibits bone morphogenetic protein-2-induced alkaline phosphatase activity in MC3T3-E1 osteoblastic cells. Bone, 21:17–21.
YamashitaM, OtsukaF, MukaiT, OtaniH, InagakiK, MiyoshiT, GotoJ, YamamuraM, MakinoH. 2008. Simvastatin antagonizes tumor necrosis factor-alpha inhibition of bone morphogenetic proteins-2-induced osteoblast differentiation by regulating Smad signaling and Ras/Rho-mitogen-activated protein kinase pathway. J Endocrinol, 196:601–613.
20.
ProckopDJ. 1997. Marrow stromal cells as steam cells for nonhematopoietic tissues. Science, 276:71–74.
21.
KaewsrichanJ, WongwitwichotP, ChandarajotiK, ChuaKH, RuszymahBH. 2011. Sequential induction of marrow stromal cells by FGF2 and BMP2 improves their growth and differentiation potential in vivo. Arch Oral Biol, 56:90–101.
ReddiAH. 1997. Bone morphogenetic proteins: an unconventional approach to isolation of first mammalian morphogens. Cytokine Growth Factor Rev, 8:11–20.
24.
LiebermanJR, DaluiskiA, EinhornTA. 2002. The role of growth factors in the repair of bone. Biology and clinical applications. J Bone Joint Surg Am, 84-A:1032–1044.
25.
TanEM, CohenAS, FriesJF, MasiAT, McShaneDJ, RothfieldNF, SchallerJG, TalalN, WinchesterRJ. 1982. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum, 25:1271–1277.
26.
SchnedES, GlicksteinSL, DoyleMA. 1993. Derivation of the SLEDAI. Arthritis Rheum, 36:877–878.
27.
SunLY, ZhangHY, FengXB, HouYY, LuLW, FanLM. 2007. Abnormality of bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus. Lupus, 16:121–128.
28.
GuoY, GuoH, ZhangL, XieH, ZhaoX, WangF, LiZ, WangY, MaSet al.2005. Genomic analysis of anti-hepatitis B virus (HBV) activity by small interfering RNA and lamivudine in stable HBV-producing cells. J Virol, 79:14392–14403.
29.
DayTF, GuoX, Garrett-BealL, YangY. 2005. Wnt/beta-catenin signaling in mesenchymal progenitors controls osteoblast and chondrocyte differentiation during vertebrate skeletogenesis. Dev Cell, 8:739–750.
30.
BaiS, KopanR, ZouW, HiltonMJ, OngCT, LongF, RossFP, TeitelbaumSL. 2008. NOTCH1 regulates osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblast lineage cells. J Biol Chem, 283:6509–6518.
31.
MishinaY, StarbuckMW, GentileMA, FukudaT, KasparcovaV, SeedorJG, HanksMC, AmlingM, PineroGJ, HaradaS, BehringerRR. 2004. Bone morphogenetic protein type IA receptor signaling regulates postnatal osteoblast function and bone remodeling. J Biol Chem, 279:27560–27566.
32.
YuK, XuJ, LiuZ, SosicD, ShaoJ, OlsonEN, TowlerDA, OrnitzDM. 2003. Conditional inactivation of FGF receptor 2 reveals an essential role for FGF signaling in the regulation of osteoblast function and bone growth. Development, 130:3063–3074.
33.
YamazakiM, FukushimaH, ShinM, KatagiriT, DoiT, TakahashiT, JimiE. 2009. Tumor necrosis factor alpha represses bone morphogenetic protein (BMP) signaling by interfering with the DNA binding of Smads through the activation of NF-kappaB. J Biol Chem, 284:35987–35995.
34.
NieY, LauCS, LieAKW, ChanGCF, MokMY. 2010. Defective phenotype of mesenchymal stem cells in patients with systemic lupus erythematosus. Lupus, 19:850–859.
35.
LiX, LiuL, MengD, WangD, ZhangJ, ShiD, LiuH, XuH, LuL, SunL. 2012. Enhanced apoptosis and senescence of bone-marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus. Stem Cells Dev, 21:2387–2394.
36.
KultererB, FriedlG, JandrositzA, Sanchez-CaboF, ProkeschA, PaarC, ScheidelerM, WindhagerR, PreiseggerKH, TrajanoskiZ. 2007. Gene expression profiling of human mesenchymal stem cells derived from bone marrow during expansion and osteoblast differentiation. BMC Genomics, 8:70.
37.
ZamurovicN, CappellenD, RohnerD, SusaM. 2004. Coordinated activation of Notch, Wnt, and transforming growth factor-beta signaling pathways in bone morphogenic protein 2-induced osteogenesis Notch-Target gene Hey1 inhibits mineralization and Runx2 transcriptional activity. J Biol Chem, 279:37704–37715.
38.
LiuT, GaoY, SakamotoK, MinamizatoT, FurukawaK, TsukazakiT, ShibataY, BesshoK, KomoriT, YamaguchiA. 2007. BMP-2 promotes differentiation of osteoblasts and chondroblasts in Runx2-deficient cell lines. J Cell Physiol, 211:728–735.
39.
DominiciM, PaolucciP, ConteP, HorwitzEM. 2009. Heterogeneity of multipotent mesenchymal stromal cells: from stromal cells to stem cells and vice versa. Transplantation, 87:S36–S42.
40.
KasperG, MaoL, GeisslerS, DraychevaA, TrippensJ, KuhnischJ, TschirschmannM, KasparK, PerkaC, DudaGN, KloseJ. 2009. Insights into mesenchymal stem cell aging: involvement of antioxidant defense and actin cytoskeleton. Stem Cells, 27:1288–1297.
41.
UeharaR, SuzuliY, IchikawaY. 2001. Methotrexate (MTX) inhibis osteoblastic differentiation in vitro: possible mechanisim of MTX osteopathy. J Rheumatol, 28:251–256.
42.
MaegawaN, KawamuraK, HiroseM, YajimaH, TakakuraY, OhgushiH. 2007. Enhancement of osteoblastic differentiation of mesenchymal stromal cells cultured by selective combination of bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor-2 (FGF-2)J Tissue Eng Regen Med, 1:306–313.
VidalNO, BrandstromH, JonssonKB, OhlssonC. 1998. Osteoprotegerin mRNA is expressed in primary human osteoblast-like cells: down-regulation by glucocorticoids. J Endocrinol, 159:191–195.
45.
MenonRK, GillDS, ThomasM, KernoffPB, DandonaP. 1987. Impaired carboxylation of osteocalcin in warfarin-treated patients. J Clin Endocrinol Metab, 64:59–61.
46.
ChengH, JiangW, PhillipsFM, HaydonRC, PengY, ZhouL, LuuHH, AnN, BreyerBet al.2003. Osteogenic activity of the fourteen types of human bone morphogenetic proteins (BMPs)J Bone Joint Surg Am, 85-A:1544–1552.
47.
LaveryK, SwainP, FalbD, Alaoui-IsmailiMH. 2008. BMP-2/4 and BMP-6/7 differentially utilize cell surface receptors to induce osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells. J Biol Chem, 283:20948–20958.
48.
LuoJ, TangM, HuangJ, HeBC, GaoJL, ChenL, ZuoGW, ZhangW, LuoQet al.2010. TGFbeta/BMP type I receptors ALK1 and ALK2 are essential for BMP9-induced osteogenic signaling in mesenchymal stem cells. J Biol Chem, 285:29588–29598.
49.
ten DijkeP. 2006. Bone morphogenetic protein signal transduction in bone. Curr Med Res Opin22 Suppl1:S7–S11.
RalstonSH, RussellRG, GowenM. 1990. Estrogen inhibits release of tumor necrosis factor from peripheral blood mononuclear cells in postmenopausal women. J Bone Miner Res, 5:983–988.
52.
PacificiR, BrownC, PuscheckE, FriedrichE, SlatopolskyE, MaggioD, McCrackenR, AvioliLV. 1991. Effect of surgical menopause and estrogen replacement on cytokine release from human blood mononuclear cells. Proc Natl Acad Sci U S A, 88:5134–5138.
53.
AmmannP, RizzoliR, BonjourJP, BourrinS, MeyerJM, VassalliP, GarciaI. 1997. Transgenic mice expressing soluble tumor necrosis factor-receptor are protected against bone loss caused by estrogen deficiency. J Clin Invest, 99:1699–16703.
54.
LiB, ShiM, LiJ, ZhangH, ChenB, ChenL, GaoW, GiulianiN, ZhaoRC. 2007. Elevated tumor necrosis factor-α suppresses TAZ expression and impairs osteogenic potential of Flk-1+ mesenchymal stem cells in patients with multiple myeloma. Stem Cells Dev, 16:921–930.
55.
KondoT, KitazawaR, YamaguchiA, KitazawaS. 2008. Dexamethasone promotes osteoclastogenesis by inhibiting osteoprotegerin through multiple levels. J Cell Biochem, 103:335–345.
56.
RubinMR, BilezikianJP. 2002. Clinical review 151: The role of parathyroid hormone in the pathogenesis of glucocorticoid-induced osteoporosis: a re-examination of the evidence. J Clin Endocrinol Metab, 87:4033–4041.
57.
WeinsteinRS, JilkaRL, ParfittAM, ManolagasSC. 1998. Inhibition of osteoblastogenesis and promotion of apoptosis of osteoblasts and osteocytes by glucocorticoids. Potential mechanisms of their deleterious effects on bone. J Clin Invest, 102:274–282.
58.
MatsumotoY, OtsukaF, TakanoM, MukaiT, YamanakaR, TakedaM, MiyoshiT, InagakiK, SadaKE, MakinoH. 2010. Estrogen and glucocorticoid regulate osteoblast differentiation through the interaction of bone morphogenetic protein-2 and tumor necrosis factor-alpha in C2C12 cells. Mol Cell Endocrinol, 325:118–127.
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