Enhancing the function of CD34(+) cells by targeting plasminogen activator inhibitor-1.
Previously, we showed that transient inhibition of TGF- β1 resulted in correction of key aspects of diabetes-induced CD34(+) cell dysfunction. In this report, we examine the effect of transient inhibition of plasminogen activator inhibitor-1 (PAI-1), a major gene target of TGF-β1 activation. Using gene array studies, we examined CD34(+) cells isolated from a cohort of longstanding diabetic individuals, free of microvascular complications despite suboptimal glycemic control, and found that the cells exhibited reduced transcripts of both TGF-β1 and PAI-1 compared to age, sex, and degree of glycemic control-matched diabetic individuals with microvascular complications. CD34(+) cells from diabetic subjects with microvascular complications consistently exhibited higher PAI-1 mRNA than age-matched non-diabetic controls. TGF- β1 phosphorodiamidate morpholino oligo (PMO) reduced PAI-1 mRNA in diabetic (p<0.01) and non-diabetic (p=0.05) CD34(+) cells. To reduce PAI-1 in human CD34(+) cells, we utilized PAI-1 siRNA, lentivirus expressing PAI-1 shRNA or PAI-1 PMO. We found that inhibition of PAI-1 promoted CD34(+) cell proliferation and migration in vitro, likely through increased PI3(K) activity and increased cGMP production. 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 PAI-1-PMO treatment compared with control PMO-treated cells. Targeting PAI-1 offers a promising therapeutic strategy for restoring vascular reparative function in defective diabetic progenitors.
Department of Pharmacology and Therapeutics, University of Florida, Gainesville, Florida, United States of America.
Sugata
S
Hazra
Valerie
V
Stepps
Ashay D
AD
Bhatwadekar
Sergio
S
Caballero
Michael E
ME
Boulton
Paul J
PJ
Higgins
Elena V
EV
Nikonova
Carl J
CJ
Pepine
Catherine
C
Thut
Eva M
EM
Finney
David J
DJ
Stone
Stephen H
SH
Bartelmez
Maria B
MB
Grant
D000328:Adult; D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism*; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation; D002478:Cells, Cultured; D015331:Cohort Studies; D006152:Cyclic GMP / Q000378:metabolism; D003920:Diabetes Mellitus / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D003925:Diabetic Angiopathies / Q000097:blood / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D017395:Plasminogen Activator Inhibitor 1 / Q000097:blood / Q000235:genetics*; D034622:RNA Interference; D015427:Reperfusion Injury / Q000378:metabolism; D012171:Retinal Vessels / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome / Q000235:genetics; D053773:Transforming Growth Factor beta1 / Q000097:blood / Q000235:genetics
D018952:Antigens, CD34; D017395:Plasminogen Activator Inhibitor 1; D053773:Transforming Growth Factor beta1; D019869:Phosphatidylinositol 3-Kinases; D006152:Cyclic GMP