Amelioration of glucose control mobilizes circulating pericyte progenitor cells in type 2 diabetic patients with microangiopathy

Gian Paolo Fadini, Patrizia Mancuso, Francesco Bertolini, Saula de Kreutzenberg, Angelo Avogaro, Gian Paolo Fadini, Patrizia Mancuso, Francesco Bertolini, Saula de Kreutzenberg, Angelo Avogaro

Abstract

Chronic diabetic complications result from an imbalance between vascular damage and regeneration. Several circulating lineage-committed progenitor cells have been implicated, but no data are available on pericyte progenitor cells (PPCs). Based on the evidence that PPCs increase in cancer patients after chemotherapy, we explored whether circulating PPC levels are affected by glucose control in type 2 diabetic patients, in relation to the presence of chronic complications. We enumerated peripheral blood PPCs as Syto16+CD45-CD31-CD140b+ events by flow cytometry at baseline and after 3 and 6 months of glucose control by means of add-on basal insulin therapy on top of oral agents in 38 poorly controlled type 2 diabetic patients. We found that, in patients with microangiopathy (n = 23), the level of circulating PPCs increased about 2 fold after 3 months and then returned to baseline at 6 months. In patients without microangiopathy (control group, n = 15), PPCs remained fairly stable during the whole study period. No relationship was found between change in PPCs and macroangiopathy (either peripheral, coronary, or cerebrovascular). We conclude that glucose control transiently mobilizes PPCs diabetic patients with microangiopathy. Increase in PPCs may represent a vasoregenerative event or may be a consequence of ameliorated glucose control on microvascular lesions.

Trial registration: ClinicalTrials.gov NCT00699686.

Figures

Figure 1
Figure 1
The gating strategy for enumeration of circulating PPCs. (a) Peripheral blood mononuclear cells were first gated into the CD45-negative fraction to exclude hematopoietic cells. (b) The total Syto16+ population of nucleated cells was selected to avoid inclusion of contaminating red cells, platelets, and debris in the analysis. (c, d) The resulting population was analyzed for expression of CD31 and CD140b. Panel (c) shows a case with low baseline PPCs (Syto16+CD45−CD31−CD140b+ cells), while (d) shows the same case 3 months after initiation of glucose control.
Figure 2
Figure 2
Effects of glucose control on HbA1c and PPCs. (a) There were no differences in HbA1c levels in patients with and without microangiopathy during time (*P < 0.05 versus baseline). (b) Increase in PPC levels was seen only in patients with microangiopathy (ANOVA P < 0.05; *post hoc P < 0.05). (c–f) Patients were divided according to the presence of micro-/macroalbuminuria, neuropathy, retinopathy, and peripheral arterial disease (PAD): a significant PPCs increase was detected in patients with urinary albumin-creatinine ratio (ACR) >30 mg/g and in the presence of neuropathy (ANOVA P < 0.05; *post hoc P < 0.05).

References

    1. Brownlee M. The pathobiology of diabetic complications: a unifying mechanism. Diabetes. 2005;54(6):1615–1625.
    1. Fadini GP, Sartore S, Agostini C, Avogaro A. Significance of endothelial progenitor cells in subjects with diabetes. Diabetes Care. 2007;30(5):1305–1313.
    1. Fadini GP. Is bone marrow another target of diabetic complications? European Journal of Clinical Investigation. 2011;41(4):457–463.
    1. Asahara T, Masuda H, Takahashi T, et al. Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circulation Research. 1999;85(3):221–228.
    1. Albiero M, Menegazzo L, Fadini GP. Circulating smooth muscle progenitors and atherosclerosis. Trends in Cardiovascular Medicine. 2010;20(4):133–140.
    1. Fadini GP, Albiero M, Menegazzo L, et al. Widespread increase in myeloid calcifying cells contributes to ectopic vascular calcification in type 2 diabetes. Circulation Research. 2011;108(9):1112–1121.
    1. Wojakowski W, Tendera M, Kucia M, et al. Cardiomyocyte differentiation of bone marrow-derived Oct-4 +CXCR4+SSEA-1+ very small embryonic-like stem cells. International Journal of Oncology. 2010;37(2):237–247.
    1. Blann AD, Woywodt A, Bertolini F, et al. Circulating endothelial cells. Biomarker of vascular disease. Thrombosis and Haemostasis. 2005;93(2):228–235.
    1. Fadini GP, Pucci L, Vanacore R, et al. Glucose tolerance is negatively associated with circulating progenitor cell levels. Diabetologia. 2007;50(10):2156–2163.
    1. Yoon YS, Uchida S, Masuo O, et al. Progressive attenuation of myocardial vascular endothelial growth factor expression is a seminal event in diabetic cardiomyopathy: restoration of microvascular homeostasis and recovery of cardiac function in diabetic cardiomyopathy after replenishment of local vascular endothelial growth factor. Circulation. 2005;111(16):2073–2085.
    1. Nguyen TQ, Chon H, Van Nieuwenhoven FA, Braam B, Verhaar MC, Goldschmeding R. Myofibroblast progenitor cells are increased in number in patients with type 1 diabetes and express less bone morphogenetic protein 6: a novel clue to adverse tissue remodelling? Diabetologia. 2006;49(5):1039–1048.
    1. McClung JA, Naseer N, Saleem M, et al. Circulating endothelial cells are elevated in patients with type 2 diabetes mellitus independently of HbA1c. Diabetologia. 2005;48(2):345–350.
    1. Mancuso P, Martin-Padura I, Calleri A, et al. Circulating perivascular progenitors: a target of PDGFR inhibition. International Journal of Cancer. 2011;129(6):1344–1350.
    1. Armulik A, Abramsson A, Betsholtz C. Endothelial/pericyte interactions. Circulation Research. 2005;97(6):512–523.
    1. Hellström M, Kalén M, Lindahl P, Abramsson A, Betsholtz C. Role of PDGF-B and PDGFR-β in recruitment of vascular smooth muscle cells and pericytes during embryonic blood vessel formation in the mouse. Development. 1999;126(14):3047–3055.
    1. Lindahl P, Johansson BR, Levéen P, Betsholtz C. Pericyte loss and microaneurysm formation in PDGF-B-deficient mice. Science. 1997;277(5323):242–245.
    1. Fadini GP, de Kreutzenberg SV, Mariano V, et al. Optimized glycaemic control achieved with add-on basal insulin therapy improves indexes of endothelial damage and regeneration in type 2 diabetic patients with macroangiopathy: a randomized crossover trial comparing detemir versus glargine. Diabetes, Obesity and Metabolism. 2011;13(8):718–725.
    1. Early Treatment Diabetic Retinopathy Study Research Group. Classification of diabetic retinopathy from fluorescein angiograms. ETDRS report number 11. Ophthalmology. 1991;98:807–822.
    1. Levey AS, Stevens LA, Schmid CH, et al. A new equation to estimate glomerular filtration rate. Annals of Internal Medicine. 2009;150(9):604–612.
    1. Wojakowski W, Ratajczak MZ, Tendera M. Mobilization of very small embryonic-like stem cells in acute coronary syndromes and stroke. Herz. 2010;35(7):467–473.
    1. Shimizu F, Sano Y, Haruki H, Kanda T. Advanced glycation end-products induce basement membrane hypertrophy in endoneurial microvessels and disrupt the blood-nerve barrier by stimulating the release of TGF-β and vascular endothelial growth factor (VEGF) by pericytes. Diabetologia. 2011;54(6):1517–1526.
    1. Kreisberg JI, Ayo SH. The glomerular mesangium in diabetes mellitus. Kidney International. 1993;43(1):109–113.
    1. Shepro D, Morel NML. Pericyte physiology. FASEB Journal. 1993;7(11):1031–1038.
    1. Mizutani M, Kern TS, Lorenzi M. Accelerated death of retinal microvascular cells in human and experimental diabetic retinopathy. Journal of Clinical Investigation. 1996;97(12):2883–2890.
    1. Fadini GP, Sartore S, Schiavon M, et al. Diabetes impairs progenitor cell mobilisation after hindlimb ischaemia-reperfusion injury in rats. Diabetologia. 2006;49(12):3075–3084.
    1. Aicher A, Rentsch M, Sasaki KI, et al. Nonbone marrow-derived circulating progenitor cells contribute to postnatal neovascularization following tissue ischemia. Circulation Research. 2007;100(4):581–589.
    1. Katare R, Riu F, Mitchell K, et al. Transplantation of human pericyte progenitor cells improves the repair of infarcted heart through activation of an angiogenic program involving micro-RNA-132. Circulation Research. 2011;109(8):894–906.
    1. Campagnolo P, Cesselli D, Al Haj Zen A, et al. Human adult vena saphena contains perivascular progenitor cells endowed with clonogenic and proangiogenic potential. Circulation. 2010;121(15):1735–1745.
    1. Grunewald M, Avraham I, Dor Y, et al. VEGF-induced adult neovascularization: recruitment, retention, and role of accessory cells. Cell. 2006;124(1):175–189.
    1. Treins C, Giorgetti-Peraldi S, Murdaca J, Semenza GL, Van Obberghen E. Insulin stimulates hypoxia-inducible factor 1 through a phosphatidylinositol 3-kinase/target of rapamycin-dependent signaling pathway. Journal of Biological Chemistry. 2002;277(31):27975–27981.
    1. Jacob A, Molkentin JD, Smolenski A, Lohmann SM, Begum N. Insulin inhibits PDGF-directed VSMC migration via NO/cGMP increase of MKP-1 and its inactivation of MAPKs. American Journal of Physiology. 2002;283(3):C704–C713.
    1. Aljada A, Friedman J, Ghanim H, et al. Glucose ingestion induces an increase in intranuclear nuclear factor κB, a fall in cellular inhibitor κB, and an increase in tumor necrosis factor α messenger RNA by mononuclear cells in healthy human subjects. Metabolism: Clinical and Experimental. 2006;55(9):1177–1185.
    1. Van Ballegooie E, Hooymans JMM, Timmerman Z. Rapid deterioration of diabetic retinopathy during treatment with continuous subcutaneous insulin infusion. Diabetes Care. 1984;7(3):236–242.
    1. Gibbons CH, Freeman R. Treatment-induced diabetic neuropathy: a reversible painful autonomic neuropathy. Annals of Neurology. 2010;67(4):534–541.
    1. Dar A, Domev H, Ben-Yosef O, et al. Multipotent vasculogenic pericytes from human pluripotent stem cells promote recovery of murine ischemic limb. Circulation. 99;2012(125):1–87.
    1. Schober A, Hristov M, Kofler S, et al. CD34+CD140b+ cells and circulating CXCL12 correlate with the angiographically assessed severity of cardiac allograft vasculopathy. European Heart Journal. 2011;32(4):476–484.

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