title: High throughput transcriptome profiling of lithium stimulated human mesenchymal stem cells reveals priming towards osteoblastic lineage.
abstract: Human mesenchymal stem cells (hMSCs) present in the bone marrow are the precursors of osteoblasts, chondrocytes and adipocytes, and hold tremendous potential for osteoregenerative therapy. However, achieving directed differentiation into osteoblasts has been a major concern. The use of lithium for enhancing osteogenic differentiation has been documented in animal models but its effect in humans is not clear. We, therefore, performed high throughput transcriptome analysis of lithium-treated hMSCs to identify altered gene expression and its relevance to osteogenic differentiation. Our results show suppression of proliferation and enhancement of alkaline phosphatase (ALP) activity upon lithium treatment of hMSCs under non-osteogenic conditions. Microarray profiling of lithium-stimulated hMSC revealed decreased expression of adipogenic genes (CEBPA, CMKLR1, HSD11B1) and genes involved in lipid biosynthesis. Interestingly, osteoclastogenic factors and immune responsive genes (IL7, IL8, CXCL1, CXCL12, CCL20) were also downregulated. Negative transcriptional regulators of the osteogenic program (TWIST1 and PBX1) were suppressed while genes involved in mineralization like CLEC3B and ATF4 were induced. Gene ontology analysis revealed enrichment of upregulated genes related to mesenchymal cell differentiation and signal transduction. Lithium priming led to enhanced collagen 1 synthesis and osteogenic induction of lithium pretreated MSCs resulted in enhanced expression of Runx2, ALP and bone sialoprotein. However, siRNA-mediated knockdown of RRAD, CLEC3B and ATF4 attenuated lithium-induced osteogenic priming, identifying a role for RRAD, a member of small GTP binding protein family, in osteoblast differentiation. In conclusion, our data highlight the transcriptome reprogramming potential of lithium resulting in higher propensity of lithium "primed" MSCs for osteoblastic differentiation.
authors: Stem Cell & Gene Therapy Research Group, Institute of Nuclear Medicine & Allied Sciences, Brig. S K Mazumdar Marg, Timarpur, Delhi, India.
authors: Neeraj Kumar
authors: NK
authors: Satija
authors: Deepa
authors: D
authors: Sharma
authors: Farhat
authors: F
authors: Afrin
authors: Rajendra P
authors: RP
authors: Tripathi
authors: Gurudutta
authors: G
authors: Gangenahalli
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D004789:Enzyme Activation / Q000187:drug effects; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D055785:Gene Knockdown Techniques; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors; D000071679:Glycogen Synthase Kinase 3 beta; D006801:Humans; D008094:Lithium / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D058977:Molecular Sequence Annotation; D010006:Osteoblasts / Q000166:cytology*; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D021381:Protein Transport / Q000187:drug effects; D015203:Reproducibility of Results; D014158:Transcription, Genetic / Q000187:drug effects; D059467:Transcriptome; D060449:Wnt Signaling Pathway / Q000187:drug effects; D051176:beta Catenin / Q000378:metabolism
chemical_list: D051176:beta Catenin; D008094:Lithium; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3; D000469:Alkaline Phosphatase
references: J Biol Chem. 2003 May 30;278(22):19674-81
references: 12668682
references: Biochem Biophys Res Commun. 2007 Jun 15;357(4):997-1003
references: 17459335
references: Biochem Biophys Res Commun. 2001 Sep 21;287(2):337-42
references: 11554731
references: Bone. 2011 Feb;48(2):321-31
references: 20932949
references: J Biol Chem. 2003 Aug 29;278(35):33067-77
references: 12796505
references: Bone. 2007 May;40(5):1252-8
references: 17258948
references: PLoS One. 2007 Sep 05;2(9):e837
references: 17786208
references: J Bone Miner Res. 2010 Feb;25(2):222-34
references: 19929432
references: Science. 2005 Aug 12;309(5737):1074-8
references: 16099986
references: BMC Syst Biol. 2010 Nov 19;4:158
references: 21092101
references: J Biol Chem. 2010 Dec 31;285(53):41310-9
references: 21044953
references: Nat Protoc. 2009;4(1):44-57
references: 19131956
references: Science. 2000 Aug 11;289(5481):950-3
references: 10937998
references: Cell Metab. 2010 Jun 9;11(6):517-31
references: 20519123
references: EMBO J. 2001 Aug 15;20(16):4500-11
references: 11500377
references: J Cell Commun Signal. 2007 Dec;1(3-4):175-83
references: 18600477
references: Neuroreport. 1999 Dec 16;10(18):3887-90
references: 10716228
references: Proc Natl Acad Sci U S A. 2007 Jun 5;104(23):9673-8
references: 17535907
references: Proc Natl Acad Sci U S A. 2009 Apr 28;106(17):7149-54
references: 19369205
references: Neuroscience. 2008 Feb 19;151(4):1184-97
references: 18222042
references: Nature. 1994 Feb 17;367(6464):651-4
references: 8107850
references: Curr Biol. 1996 Dec 1;6(12):1664-8
references: 8994831
references: Stem Cells. 2004;22(5):823-31
references: 15342946
references: Bone. 2004 May;34(5):818-26
references: 15121013
references: Cell. 2004 Apr 30;117(3):387-98
references: 15109498
references: PLoS One. 2007 Sep 26;2(9):e945
references: 17895986
references: Genes Dev. 2001 Dec 1;15(23):3088-103
references: 11731474
references: J Proteome Res. 2010 Jun 4;9(6):2946-56
references: 20359165
references: Ann Intern Med. 1990 Mar 1;112(5):352-64
references: 2407167
references: Stem Cells. 2002;20(6):530-41
references: 12456961
references: Transgenic Res. 2010 Dec;19(6):1017-39
references: 20174869
references: Proc Natl Acad Sci U S A. 2005 Nov 29;102(48):17406-11
references: 16293698
references: Stem Cell Rev Rep. 2009 Dec;5(4):378-86
references: 20058201
references: BMC Bioinformatics. 2009 Feb 03;10:45
references: 19192265
references: Int J Biochem Cell Biol. 2011 May;43(5):760-7
references: 21296678
references: J Cell Biol. 2002 Apr 15;157(2):291-302
references: 11956230
references: Am J Physiol Renal Physiol. 2005 Apr;288(4):F703-13
references: 15572521
references: J Endocrinol. 2008 May;197(2):297-307
references: 18434359
references: Trends Biochem Sci. 2010 Mar;35(3):161-8
references: 19884009
references: Genes Cells. 2009 Mar;14(3):407-24
references: 19228201
references: Nat Genet. 2002 Dec;32 Suppl:490-5
references: 12454643
references: Stem Cells. 2009 Oct;27(10):2457-68
references: 19609939
references: Endocrinology. 2004 Apr;145(4):1916-25
references: 14715714
references: Bone. 2009 Feb;44(2):331-4
references: 18992857
references: J Biol Chem. 2008 Mar 7;283(10):6241-52
references: 18178562
references: Calcif Tissue Int. 2005 Jul;77(1):1-8
references: 16007481
references: Bone. 2010 Apr;46(4):985-92
references: 20060080
references: Stem Cells. 2008 Sep;26(9):2399-407
references: 18583541
references: Tissue Eng. 2004 Mar-Apr;10(3-4):393-401
references: 15165456
references: J Dent Res. 2011 Mar;90(3):336-40
references: 21118797
references: Stem Cells. 2011 Jul;29(7):1102-11
references: 21608080
references: Front Biosci. 1998 Aug 01;3:d804-16
references: 9682034
references: J Bone Miner Res. 2004 Apr;19(4):661-70
references: 15005854
references: J Bone Miner Res. 2003 Oct;18(10):1842-53
references: 14584895
references: N Engl J Med. 2001 Feb 1;344(5):385-6
references: 11195802
references: Trends Cell Biol. 2006 Mar;16(3):151-8
references: 16466918
references: J Clin Psychopharmacol. 2008 Aug;28(4):384-91
references: 18626264
references: Oral Dis. 2007 Jan;13(1):32-9
references: 17241427
references: Stem Cells. 2005 Oct;23(9):1357-66
references: 16081661
references: J Cell Biol. 1994 Dec;127(6 Pt 1):1767-75
references: 7798325
references: Bioinformatics. 2001 Jun;17(6):509-19
references: 11395427
references: BMC Cell Biol. 2008 Feb 13;9:11
references: 18271953
references: Exp Hematol. 2008 Sep;36(9):1176-85
references: 18495329
references: Biochem Biophys Res Commun. 2003 Jan 31;301(1):84-91
references: 12535644
references: Bioinformatics. 2004 Sep 1;20(13):2156-8
references: 15145817
references: EMBO J. 1998 Apr 1;17(7):2019-32
references: 9524124
references: J Cell Biol. 2009 Apr 6;185(1):67-75
references: 19349579
references: EMBO J. 1999 Mar 1;18(5):1270-9
references: 10064593
references: J Bone Miner Res. 2010 Apr;25(4):796-808
references: 19821776
references: Dev Cell. 2004 Mar;6(3):423-35
references: 15030764
references: J Cell Mol Med. 2009 Nov-Dec;13(11-12):4385-402
references: 19602034
references: Endocrinology. 2003 Sep;144(9):4134-43
references: 12933688
references: Stem Cells Dev. 2007 Feb;16(1):7-23
references: 17348802
references: Proc Natl Acad Sci U S A. 2002 Jun 25;99(13):8932-7
references: 12084934
references: Bone. 2006 Jan;38(1):21-9
references: 16126465
references: Mol Cell Biol. 2010 Jul;30(14):3531-41
references: 20439491
references: Am J Physiol Endocrinol Metab. 2009 Jan;296(1):E139-46
references: 19001547
references: J Biol Chem. 2003 Jul 25;278(30):28067-78
references: 12740383
references: Stem Cells. 2008 Jan;26(1):193-201
references: 17916801
references: Trends Biotechnol. 2005 Aug;23(8):429-35
references: 15950303
references: J Biol Chem. 2004 Nov 5;279(45):47109-14
references: 15377660
references: Blood. 2005 May 15;105(10):3793-801
references: 15677562
references: Stem Cells. 2006 Aug;24(8):1892-903
references: 16690780
references: Apoptosis. 2005 Jan;10(1):75-90
references: 15711924
references: J Clin Invest. 2002 Jun;109(11):1405-15
references: 12045254
references: J Biol Chem. 2005 Jan 21;280(3):2309-23
references: 15504735
references: Proc Natl Acad Sci U S A. 2010 Mar 2;107(9):4147-52
references: 20150512
references: J Cell Biol. 2007 Feb 26;176(5):709-18
references: 17325210
references: J Biol Chem. 2007 May 11;282(19):14515-24
references: 17351296
references: Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):7841-5
references: 11427725