Transient inhibition of transforming growth factor-beta1 in human diabetic CD34+ cells enhances vascular reparative functions.
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.
Bhatwadekar
Ashay D
AD
Pharmacology and Therapeutics, University of Florida, Gainesville, Florida, USA.
Guerin
E P
EP
Jarajapu
Yagna P R
YP
Caballero
Sergio
S
Sheridan
Carl
C
Kent
David
D
Kennedy
Laurence
L
Lansang
M Cecilia
MC
Ruscetti
Frank W
FW
Pepine
Carl J
CJ
Higgins
Paul J
PJ
Bartelmez
Stephen H
SH
Grant
Maria B
MB
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*
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
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