Restricted accessResearch articleFirst published online 2010-10
Identification of a Common Gene Expression Signature Associated with Immature Clonal Mesenchymal Cell Populations Derived from Bone Marrow and Dental Tissues
Mesenchymal stem/stromal cell-like populations derived from adult bone marrow (BMSC), dental pulp (DPSC), and periodontal ligament (PDLSC) have the ability to differentiate into cells of mesenchymal and non-mesenchymal tissues in vitro and in vivo. However, culture-expanded MSC-like populations are a heterogeneous mix of stem/committed progenitor cells that exhibit altered growth and developmental potentials. In the present study we isolated and characterized clonal populations of BMSCs, DPSCs, and PDLSCs to identify potential biomarkers associated with long-lived multipotential stem cells. Microarray analysis was used to compare the global gene expression profiles of high growth/multipotential clones with low growth potential cell clones derived from 3 stromal tissues. Cross-comparison analyses of genes expressed by high growth/multipotential clones derived from bone marrow, dental pulp, and periodontal ligament identified 24 genes that are differentially up-regulated in all tissues. Notably, the transcription factors, E2F2, PTTG1, TWIST-1, and transcriptional cofactor, LDB2, each with critical roles in cell growth and survival, were highly expressed in all stem cell populations examined. These findings provide a model system for identifying a common molecular fingerprint associated with immature mesenchymal stem-like cells from different organs and implicate a potential role for these genes in MSC growth and development.
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References
1.
FriedensteinAJ, GorskajaJF, KulaginaNN. 1976. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol, 4:267–274.
2.
FriedensteinAJ, Piatetzky-ShapiroII, PetrakovaKV. 1966. Osteogenesis in transplants of bone marrow cells. J Embryol Exp Morphol, 16:381–390.
3.
Castro-MalaspinaH, GayRE, ResnickG, KapoorN, MeyersP, ChiarieriD, McKenzieS, BroxmeyerHE, MooreMA. 1980. Characterization of human bone marrow fibroblast colony-forming cells (CFU-F) and their progeny. Blood, 56:289–301.
4.
HaynesworthSE, GoshimaJ, GoldbergVM, CaplanAI. 1992. Characterization of cells with osteogenic potential from human marrow. Bone, 13:81–88.
5.
KuznetsovSA, KrebsbachPH, SatomuraK, KerrJ, RiminucciM, BenayahuD, RobeyPG. 1997. Single-colony derived strains of human marrow stromal fibroblasts form bone after transplantation in vivo. J Bone Miner Res, 12:1335–1347.
6.
GronthosS, ZannettinoAC, HaySJ, ShiS, GravesSE, KortesidisA, SimmonsPJ. 2003. Molecular and cellular characterisation of highly purified stromal stem cells derived from human bone marrow. J Cell Sci, 116,Pt 9:1827–1835.
7.
GronthosS, MankaniM, BrahimJ, RobeyPG, ShiS. 2000. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci USA, 97:13625–13630.
8.
SeoBM, MiuraM, GronthosS, BartoldPM, BatouliS, BrahimJ, YoungM, RobeyPG, WangCY, ShiS. 2004. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet, 364:149–155.
9.
MiuraM, GronthosS, ZhaoM, LuB, FisherLW, RobeyPG, ShiS. 2003. SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci USA, 100:5807–5812.
10.
ShiS, RobeyPG, GronthosS. 2001. Comparison of human dental pulp and bone marrow stromal stem cells by cDNA microarray analysis. Bone, 29:532–539.
W rightGW, SimonRM. 2003. A random variance model for detection of differential gene expression in small microarray experiments. Bioinformatics, 19:2448–2455.
15.
ShiS, GronthosS. 2003. Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp. J Bone Miner Res, 18:696–704.
16.
ArthurA, RychkovG, ShiS, KoblarSA, GronthosS. 2008. Adult human dental pulp stem cells differentiate toward functionally active neurons under appropriate environmental cues. Stem Cells, 26:1787–1795.
17.
SimmonsPJ, Torok-StorbB. 1991. Identification of stromal cell precursors in human bone marrow by a novel monoclonal antibody, STRO-1. Blood, 78:55–62.
18.
GronthosS, GravesSE, OhtaS, SimmonsPJ. 1994. The STRO-1+ fraction of adult human bone marrow contains the osteogenic precursors. Blood, 84:4164–4173.
AkaviaUD, VeinblatO, BenayahuD. 2008. Comparing the transcriptional profile of mesenchymal cells to cardiac and skeletal muscle cells. J Cell Physiol, 216:663–672.
25.
CamH, DynlachtBD. 2003. Emerging roles for E2F: beyond the G1/S transition and DNA replication. Cancer Cell, 3:311–316.
26.
DeGregoriJ. 2002. The genetics of the E2F family of transcription factors: shared functions and unique roles. Biochim Biophys Acta, 1602:131–150.
27.
StevauxO, DysonNJ. 2002. A revised picture of the E2F transcriptional network and RB function. Curr Opin Cell Biol, 14:684–691.
28.
TimmersC, SharmaN, OpavskyR, MaitiB, WuL, WuJ, OrringerD, TrikhaP, SaavedraHI, LeoneG. 2007. E2f1, E2f2, and E2f3 control E2F target expression and cellular proliferation via a p53-dependent negative feedback loop. Mol Cell Biol, 27:65–78.
29.
TrimarchiJM, LeesJA. 2002. Sibling rivalry in the E2F family. Nat Rev Mol Cell Biol, 3:11–20.
30.
WonJ, YimJ, KimTK. 2002. Opposing regulatory roles of E2F in human telomerase reverse transcriptase (hTERT) gene expression in human tumor and normal somatic cells. FASEB J, 16:1943–1945.
31.
ShiS, GronthosS, ChenS, ReddiA, CounterCM, RobeyPG, WangCY. 2002. Bone formation by human postnatal bone marrow stromal stem cells is enhanced by telomerase expression. Nat Biotechnol, 20:587–591.
32.
GronthosS, ChenS, WangCY, RobeyPG, ShiS. 2003. Telomerase accelerates osteogenesis of bone marrow stromal stem cells by upregulation of CBFA1, osterix, and osteocalcin. J Bone Miner Res, 18:716–722.
33.
SimonsenJL, RosadaC, SerakinciN, JustesenJ, StenderupK, RattanSI, JensenTG, KassemM. 2002. Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells. Nat Biotechnol, 20:592–596.
34.
PeiL, MelmedS. 1997. Isolation and characterization of a pituitary tumor-transforming gene (P TTG)Mol Endocrinol, 11:433–441.
35.
ZouH, McGarryTJ, BernalT, KirschnerMW. 1999. Identification of a vertebrate sister-chromatid separation inhibitor involved in transformation and tumorigenesis. Science, 285:418–422.
36.
JallepalliPV, WaizeneggerIC, BunzF, LangerS, SpeicherMR, PetersJM, KinzlerKW, VogelsteinB, LengauerC. 2001. Securin is required for chromosomal stability in human cells. Cell, 105:445–457.
37.
Ramos-MoralesF, DominguezA, RomeroF, LunaR, MultonMC, Pintor-ToroJA, TortoleroM. 2000. Cell cycle regulated expression and phosphorylation of hpttg proto-oncogene product. Oncogene, 19:403–409.
38.
RomeroF, MultonMC, Ramos-MoralesF, DomínguezA, BernalJA, Pintor-ToroJA, TortoleroM. 2001. Human securin, hPTTG, is associated with Ku heterodimer, the regulatory subunit of the DNA-dependent protein kinase. Nucleic Acids Res, 29:1300–1307.
39.
ChamaonK, KirchesE, KanakisD, BraeuningerS, DietzmannK, MawrinC. 2005. Regulation of the pituitary tumor transforming gene by insulin-like-growth factor-I and insulin differs between malignant and non-neoplastic astrocytes. Biochem Biophys Res Commun, 331:86–92.
40.
HeaneyAP, NelsonV, FernandoM, HorwitzG. 2001. Transforming events in thyroid tumorigenesis and their association with follicular lesions. J Clin Endocrinol Metab, 86:5025–5032.
41.
HeaneyAP, SingsonR, McCabeCJ, NelsonV, NakashimaM, MelmedS. 2000. Expression of pituitary-tumour transforming gene in colorectal tumours. Lancet, 355:716–719.
42.
KakarSS, MalikMT. 2006. Suppression of lung cancer with siRNA targeting PTTG. Int J Oncol, 29:387–395.
43.
PuriR, ToussonA, ChenL, KakarSS. 2001. Molecular cloning of pituitary tumor transforming gene 1 from ovarian tumors and its expression in tumors. Cancer Lett, 163:131–139.
44.
SolbachC, RollerM, FellbaumC, NicolettiM, KaufmannM. 2004. PTTG mRNA expression in primary breast cancer: a prognostic marker for lymph node invasion and tumor recurrence. Breast, 13:80–81.
45.
TsaiSJ, LinSJ, ChengYM, ChenHM, WingLY. 2005. Expression and functional analysis of pituitary tumor transforming gene-1 [corrected] in uterine leiomyomas. J Clin Endocrinol Metab, 90:3715–3723.
MeiJ, HuangX, ZhangP. 2001. Securin is not required for cellular viability, but is required for normal growth of mouse embryonic fibroblasts. Curr Biol, 11:1197–1201.
48.
RubinekT, ChesnokovaV, WolfI, WawrowskyK, VlotidesG, MelmedS. 2007. Discordant proliferation and differentiation in pituitary tumor-transforming gene-null bone marrow stem cells. Am J Physiol, Cell Physiol, 293:C1082–C1092.
49.
ChenZF, BehringerRR. 1995. twist is required in head mesenchyme for cranial neural tube morphogenesis. Genes Dev, 9:686–699.
50.
LeeMS, LoweGN, StrongDD, WergedalJE, GlackinCA. 1999. TWIST, a basic helix-loop-helix transcription factor, can regulate the human osteogenic lineage. J Cell Biochem, 75:566–577.
51.
LeptinM. 1991. twist and snail as positive and negative regulators during Drosophila mesoderm development. Genes Dev, 5:1568–1576.
52.
LeeMS, LoweG, FlanaganS, KuchlerK, GlackinCA. 2000. Human Dermo-1 has attributes similar to twist in early bone development. Bone, 27:591–602.
53.
BurgessR, RawlsA, BrownD, BradleyA, OlsonEN. 1996. Requirement of the paraxis gene for somite formation and musculoskeletal patterning. Nature, 384:570–573.
54.
CserjesiP, BrownD, LigonKL, LyonsGE, CopelandNG, GilbertDJ, JenkinsNA, OlsonEN. 1995. Scleraxis: a basic helix-loop-helix protein that prefigures skeletal formation during mouse embryogenesis. Development, 121:1099–1110.
55.
AlborziA, MacK, GlackinCA, MurraySS, ZernikJH. 1996. Endochondral and intramembranous fetal bone development: osteoblastic cell proliferation, and expression of alkaline phosphatase, m-twist, and histone H4. J Craniofac Genet Dev Biol, 16:94–106.
56.
ConnerneyJ, AndreevaV, LeshemY, MuentenerC, MercadoMA, SpicerDB. 2006. Twist1 dimer selection regulates cranial suture patterning and fusion. Dev Dyn, 235:1345–1357.
57.
RiceDP, AbergT, ChanY, TangZ, KettunenPJ, PakarinenL, MaxsonRE, ThesleffI. 2000. Integration of FGF and TWIST in calvarial bone and suture development. Development, 127:1845–1855.
58.
BialekP, KernB, YangX, SchrockM, SosicD, HongN, WuH, YuK, OrnitzDM, OlsonEN, JusticeMJ, KarsentyG. 2004. A twist code determines the onset of osteoblast differentiation. Dev Cell, 6:423–435.
59.
KronenbergHM. 2004. Twist genes regulate Runx2 and bone formation. Dev Cell, 6:317–318.
60.
RiceDP, RiceR, ThesleffI. 2003. Molecular mechanisms in calvarial bone and suture development, and their relation to craniosynostosis. Eur J Orthod, 25:139–148.
61.
FunatoN, OhtaniK, OhyamaK, KurodaT, NakamuraM. 2001. Common regulation of growth arrest and differentiation of osteoblasts by helix-loop-helix factors. Mol Cell Biol, 21:7416–7428.
62.
MurraySS, GlackinCA, WintersKA, GazitD, KahnAJ, MurrayEJ. 1992. Expression ofhelix-loop-helix regulatory genes during differentiation of mouse osteoblastic cells. J Bone Miner Res, 7:1131–1138.
63.
TamuraM, NodaM. 1999. Identification of DERMO-1 as a member of helix-loop-helix type transcription factors expressed in osteoblastic cells. J Cell Biochem, 72:167–176.
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