title: Transient inhibition of transforming growth factor-beta1 in human diabetic CD34+ cells enhances vascular reparative functions.
abstract: OBJECTIVE
  Peripheral blood CD34(+) cells from diabetic patients demonstrate reduced vascular reparative function due to decreased proliferation and diminished migratory prowess, largely resulting from decreased nitric oxide (NO) bioavailability. The level of TGF-beta, a key factor that modulates stem cell quiescence, is increased in the serum of type 2 diabetic patients. We asked whether transient TGF-beta1 inhibition in CD34(+) cells would improve their reparative ability.
  
  
  METHODS
  To inhibit TGF-beta1 protein expression, CD34(+) cells were treated ex vivo with antisense phosphorodiamidate morpholino oligomers (TGF-beta1-PMOs) and analyzed for cell surface CXCR4 expression, cell survival in the absence of added growth factors, SDF-1-induced migration, NO release, and in vivo retinal vascular reparative ability.
  
  
  RESULTS
  TGF-beta1-PMO treatment of diabetic CD34(+) cells resulted in increased expression of CXCR4, enhanced survival in the absence of growth factors, and increased migration and NO release as compared with cells treated with control PMO. Using a retinal ischemia reperfusion injury model in mice, we observed that recruitment of diabetic CD34(+) cells to injured acellular retinal capillaries was greater after TGF-beta1-PMO treatment compared with control PMO-treated cells.
  
  
  CONCLUSIONS
  Transient inhibition of TGF-beta1 may represent a promising therapeutic strategy for restoring the reparative capacity of dysfunctional diabetic CD34(+) cells.
authors.affiliation: Pharmacology and Therapeutics, University of Florida, Gainesville, Florida, USA.
authors.forename: Ashay D
authors.initials: AD
authors.lastname: Bhatwadekar
authors.forename: E P
authors.initials: EP
authors.lastname: Guerin
authors.forename: Yagna P R
authors.initials: YP
authors.lastname: Jarajapu
authors.forename: Sergio
authors.initials: S
authors.lastname: Caballero
authors.forename: Carl
authors.initials: C
authors.lastname: Sheridan
authors.forename: David
authors.initials: D
authors.lastname: Kent
authors.forename: Laurence
authors.initials: L
authors.lastname: Kennedy
authors.forename: M Cecilia
authors.initials: MC
authors.lastname: Lansang
authors.forename: Frank W
authors.initials: FW
authors.lastname: Ruscetti
authors.forename: Carl J
authors.initials: CJ
authors.lastname: Pepine
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Higgins
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Bartelmez
authors.forename: Maria B
authors.initials: MB
authors.lastname: Grant
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism / Q000502:physiology; D002196:Capillaries / Q000503:physiopathology; D002470:Cell Survival; D003920:Diabetes Mellitus / Q000503:physiopathology*; D003925:Diabetic Angiopathies / Q000517:prevention & control*; D003930:Diabetic Retinopathy / Q000503:physiopathology; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D051379:Mice; D009025:Morpholines / Q000494:pharmacology / Q000627:therapeutic use; D060172:Morpholinos; D009569:Nitric Oxide / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000235:genetics; D015427:Reperfusion Injury / Q000503:physiopathology; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D018952:Antigens, CD34; C527276:CXCR4 protein, human; D009025:Morpholines; D060172:Morpholinos; D019718:Receptors, CXCR4; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D009569:Nitric Oxide
references.citation: J Am Coll Cardiol. 2005 May 3;45(9):1449-57
references.pmid: 15862417
references.citation: Science. 1997 Feb 14;275(5302):964-7
references.pmid: 9020076
references.citation: Stem Cells. 2009 Nov;27(11):2857-64
references.pmid: 19711453
references.citation: Stem Cells. 1993 Jul;11(4):336-47
references.pmid: 7691312
references.citation: Eur J Haematol. 2003 Nov;71(5):351-8
references.pmid: 14667198
references.citation: Curr Top Med Chem. 2007;7(7):651-60
references.pmid: 17430206
references.citation: Oncogene. 2005 Aug 29;24(37):5751-63
references.pmid: 16123808
references.citation: Nat Med. 2002 Jun;8(6):607-12
references.pmid: 12042812
references.citation: Diabetologia. 2008 Jul;51(7):1296-305
references.pmid: 18286257
references.citation: J Vasc Surg. 2009 Dec;50(6):1412-22
references.pmid: 19837544
references.citation: Blood. 2001 Feb 1;97(3):799-804
references.pmid: 11157500
references.citation: Nat Med. 2003 Nov;9(11):1370-6
references.pmid: 14556003
references.citation: Blood. 2002 Jan 15;99(2):499-506
references.pmid: 11781230
references.citation: Circ Res. 2009 Oct 23;105(9):897-905
references.pmid: 19762684
references.citation: Diabetes Res Clin Pract. 2008 Jul;81(1):25-32
references.pmid: 18485515
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 19;104(25):10595-600
references.pmid: 17567755
references.citation: Nature. 1999 Jun 10;399(6736):601-5
references.pmid: 10376603
references.citation: Blood. 2002 May 1;99(9):3454-7
references.pmid: 11964317
references.citation: Invest Ophthalmol Vis Sci. 2005 Jan;46(1):343-8
references.pmid: 15623794
references.citation: Diabetes. 2006 Jan;55(1):102-9
references.pmid: 16380482
references.citation: J Immunol. 1999 Dec 1;163(11):5954-63
references.pmid: 10570282
references.citation: Blood. 2010 Apr 8;115(14):2818-26
references.pmid: 20130240
references.citation: J Cell Physiol. 2009 May;219(2):235-42
references.pmid: 19115244
references.citation: Exp Neurol. 2008 Jun;211(2):469-79
references.pmid: 18406405
references.citation: Circ Res. 2001 Feb 2;88(2):167-74
references.pmid: 11157668
references.citation: J Exp Med. 1988 Aug 1;168(2):737-50
references.pmid: 3261777
references.citation: Invest Ophthalmol Vis Sci. 2007 Jan;48(1):361-7
references.pmid: 17197555
references.citation: Diabetes. 2008 Sep;57(9):2488-94
references.pmid: 18559661
references.citation: Invest Ophthalmol Vis Sci. 2003 Nov;44(11):4908-13
references.pmid: 14578416
references.citation: J Exp Med. 2009 Dec 21;206(13):2897-906
references.pmid: 19934019
references.citation: Arch Ophthalmol. 2002 Dec;120(12):1644-50
references.pmid: 12470137
references.citation: Diabetologia. 2006 Dec;49(12):3075-84
references.pmid: 17072586
references.citation: J Immunol. 2001 Mar 1;166(5):3067-74
references.pmid: 11207257
references.citation: J Thromb Haemost. 2009 Jul;7 Suppl 1:31-4
references.pmid: 19630763
references.citation: J Clin Invest. 2000 Aug;106(4):571-8
references.pmid: 10953032
references.citation: Exp Hematol. 2001 Sep;29(9):1098-108
references.pmid: 11532351
references.citation: J Exp Med. 2002 May 20;195(10):1247-55
references.pmid: 12021305
references.citation: J Biol Chem. 2008 Apr 18;283(16):10930-8
references.pmid: 18227068
references.citation: FASEB J. 2007 Oct;21(12):3219-30
references.pmid: 17522382
references.citation: Blood. 1996 Jul 1;88(1):82-8
references.pmid: 8704205
references.citation: Diabetes Care. 2009 Oct;32(10):1921-3
references.pmid: 19592635
references.citation: Diabetes. 2007 Apr;56(4):960-7
references.pmid: 17395742
references.citation: Nature. 1999 Jun 10;399(6736):597-601
references.pmid: 10376602
references.citation: Curr Top Dev Biol. 2004;64:141-80
references.pmid: 15563947
references.citation: Int J Hematol. 2001 Jul;74(1):18-25
references.pmid: 11530800
references.citation: Proc Natl Acad Sci U S A. 1998 Oct 27;95(22):13006-11
references.pmid: 9789031
references.citation: Exp Gerontol. 2003 Oct;38(10):1179-88
references.pmid: 14580871