title: The Casitas B lineage lymphoma (Cbl) mutant G306E enhances osteogenic differentiation in human mesenchymal stromal cells in part by decreased Cbl-mediated platelet-derived growth factor receptor alpha and fibroblast growth factor receptor 2 ubiquitination.
abstract: Human bone marrow-derived mesenchymal stromal cells (hMSCs) have the capacity to differentiate into several cell types including osteoblasts and are therefore an important cell source for bone tissue regeneration. A crucial issue is to identify mechanisms that trigger hMSC osteoblast differentiation to promote osteogenic potential. Casitas B lineage lymphoma (Cbl) is an E3 ubiquitin ligase that ubiquitinates and targets several molecules for degradation. We hypothesized that attenuation of Cbl-mediated degradation of receptor tyrosine kinases (RTKs) may promote osteogenic differentiation in hMSCs. We show here that specific inhibition of Cbl interaction with RTKs using a Cbl mutant (G306E) promotes expression of osteoblast markers (Runx2, alkaline phosphatase, type 1 collagen, osteocalcin) and increases osteogenic differentiation in clonal bone marrow-derived hMSCs and primary hMSCs. Analysis of molecular mechanisms revealed that the Cbl mutant increased PDGF receptor α and FGF receptor 2 but not EGF receptor expression in hMSCs, resulting in increased ERK1/2 and PI3K signaling. Pharmacological inhibition of FGFR or PDGFR abrogated in vitro osteogenesis induced by the Cbl mutant. The data reveal that specific inhibition of Cbl interaction with RTKs promotes the osteogenic differentiation program in hMSCs in part by decreased Cbl-mediated PDGFRα and FGFR2 ubiquitination, providing a novel mechanistic approach targeting Cbl to promote the osteogenic capacity of hMSCs.
authors: Laboratory of Osteoblast Biology and Pathology, INSERM, U606, Paris F-75475, France.
authors: Nicolas
authors: N
authors: Sévère
authors: Hichem
authors: H
authors: Miraoui
authors: Pierre J
authors: PJ
authors: Marie
references: Sci Signal. 2009 Dec 22;2(102):ra86
references: 20029031
references: Nat Rev Mol Cell Biol. 2005 Dec;6(12):907-18
references: 16227975
references: Stem Cells. 2001;19(3):180-92
references: 11359943
references: J Biol Chem. 2004 Aug 27;279(35):36259-67
references: 15190072
references: Cloning Stem Cells. 2004;6(4):369-74
references: 15671665
references: Trends Biochem Sci. 2006 Feb;31(2):79-88
references: 16406635
references: Crit Rev Eukaryot Gene Expr. 2004;14(1-2):1-41
references: 15104525
references: J Biol Chem. 1997 Dec 26;272(52):33140-4
references: 9407100
references: Blood. 2007 Jul 1;110(1):334-8
references: 17371942
references: Am J Pathol. 2006 Oct;169(4):1303-11
references: 17003487
references: Gene. 2003 Oct 16;316:23-32
references: 14563548
references: Oncogene. 1997 May 8;14(18):2239-49
references: 9174060
references: Cancer Res. 2007 Apr 15;67(8):3708-15
references: 17440083
references: J Biol Chem. 2000 Mar 31;275(13):9645-52
references: 10734116
references: J Biol Chem. 1996 Sep 27;271(39):24063-8
references: 8798643
references: Mol Cell Biol. 1997 Aug;17(8):4597-610
references: 9234717
references: Platelets. 2004 Feb;15(1):29-35
references: 14985174
references: J Biol Chem. 1999 Jun 4;274(23):16619-28
references: 10347229
references: J Bone Miner Res. 1996 Feb;11(2):238-47
references: 8822348
references: J Bone Miner Res. 1999 Mar;14(3):351-61
references: 10027900
references: Proc Natl Acad Sci U S A. 2003 Sep 30;100 Suppl 1:11917-23
references: 13679583
references: J Biol Chem. 2003 Aug 29;278(35):33456-64
references: 12815057
references: Endocrinology. 1994 Jan;134(1):277-86
references: 8275945
references: Gene Ther. 2001 Feb;8(3):190-8
references: 11313790
references: Trends Cell Biol. 2006 Jan;16(1):45-54
references: 16337795
references: Bone. 2008 Jun;42(6):1032-9
references: 18374639
references: J Cell Biochem. 2001;81(1):23-38
references: 11180395
references: Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):7927-32
references: 9653117
references: J Clin Invest. 2004 Nov;114(9):1326-33
references: 15520865
references: J Bone Miner Res. 2010 Oct;25(10):2115-25
references: 20499359
references: Hum Mol Genet. 2010 May 1;19(9):1678-89
references: 20124286
references: J Biol Chem. 1997 Mar 28;272(13):8739-43
references: 9079708
references: Nat Rev Mol Cell Biol. 2001 Apr;2(4):294-307
references: 11283727
references: J Biol Chem. 2009 Feb 20;284(8):4897-904
references: 19117954
references: J Cell Biochem. 2003 Feb 15;88(3):446-54
references: 12532321
references: J Biol Chem. 2004 Jul 2;279(27):28017-27
references: 15117950
references: J Biol Chem. 2003 May 30;278(22):20091-7
references: 12649282
references: Crit Rev Oral Biol Med. 1999;10(1):40-57
references: 10759426
references: Curr Top Microbiol Immunol. 1990;166:159-64
references: 2073794
references: Stem Cells Dev. 2007 Dec;16(6):953-63
references: 18158854
references: Stem Cell Rev Rep. 2011 Mar;7(1):32-42
references: 20198518
references: J Clin Invest. 2008 Feb;118(2):491-504
references: 18219387
references: Science. 2005 Jun 3;308(5727):1472-7
references: 15933201
references: Mol Cell Biol. 2009 Feb;29(3):881-91
references: 19047372
references: EMBO J. 2002 Sep 16;21(18):4796-808
references: 12234920
references: J Cell Sci. 2005 Mar 15;118(Pt 6):1223-32
references: 15728256
references: J Bone Miner Res. 2006 May;21(5):722-34
references: 16734387
references: J Biol Chem. 1999 Aug 6;274(32):22151-4
references: 10428778
references: J Clin Invest. 2003 Jun;111(11):1771-82
references: 12782679
references: J Bone Miner Res. 2001 Sep;16(9):1600-10
references: 11547830
references: Crit Rev Oncog. 1997;8(2-3):189-218
references: 9570294
references: J Cell Physiol. 2006 Oct;209(1):21-43
references: 16741904
references: Science. 2004 Nov 26;306(5701):1506-7
references: 15567848
references: Sci STKE. 2001 Nov 27;2001(110):pe40
references: 11724969
references: J Bone Miner Res. 2006 Apr;21(4):637-46
references: 16598384
references: Biochem Soc Trans. 2009 Dec;37(Pt 6):1189-92
references: 19909244
references: Regen Med. 2006 Jul;1(4):539-48
references: 17465848
references: Sci Signal. 2010 Nov 02;3(146):re9
references: 21045207
references: Circ Res. 2004 Jul 9;95(1):9-20
references: 15242981
references: Cell Death Differ. 2006 Nov;13(11):1845-56
references: 16470222
references: Proc Natl Acad Sci U S A. 2002 May 14;99(10):6684-9
references: 11997436