Abstract
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.
Highlights
With the global pandemic of diabetes affecting every continent, the impact of diabetic micro- and macro-vascular complications is far reaching
We hypothesized that diabetic individuals protected from the development of microvascular complications might have more robust CD34+ cell function with a superior reparative response compared to CD34+ cells from diabetic individuals manifesting vascular complications
We compared the CD34+ cells from protected diabetic individuals to diabetic individuals with microvascular complications that were matched for sex, age and glucose control, as well as to healthy controls using Affymetrix microarrays (Table 1). 270 probe sets were found differentially expressed with p
Summary
With the global pandemic of diabetes affecting every continent, the impact of diabetic micro- and macro-vascular complications is far reaching. Transient downregulation and functional inhibition of the intracellular TGF-β1 pathway in diabetic human CD34+ cells corrects key aspects of their dysfunctional behavior [12] and this likely occurs through effects on critical TGF-β1 target genes. To this end, recent data confirms the role of one such TGF-β1-regulated gene, PAI-1 (SERPINE1), as an important mediator of cellular growth arrest [13]. CD34+ cells express low-density lipoprotein receptor-related protein 1(LRP-1), the putative receptor for PAI-1 [26], supporting that PAI-1 may mediate both paracrine and autocrine effects on CD34+ cells
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