[show abstract][hide abstract] ABSTRACT: MicroRNAs are small noncoding RNAs that posttranscriptionally control gene expression by targeting mRNAs. Distinct microRNAs regulate stem and progenitor cell functions, thereby modulating cell survival and homing or controlling differentiation and maturation. Experimental studies additionally show that microRNAs regulate endogenous repair and might potentially be useful to enhance the regeneration of the heart. This review summarizes the current studies that address the use of microRNAs to either improve cellular therapies or that might be targeted for enhancing endogenous tissue repair and regeneration after myocardial infarction.
[show abstract][hide abstract] ABSTRACT: MicroRNAs (miRs) are small non-coding RNAs that posttranscriptionally control gene expression. Small animal studies suggest that miRs might offer novel therapeutic targets in cardiovascular disease, such as cardioprotection of murine hearts after myocardial infarction via miR-92a inhibitors. Since the functional benefits of miR-92a inhibitors in larger pre-clinical models are not known, we assessed the therapeutic efficacy of miR-92a inhibition in a porcine model of ischemia and reperfusion.
Pigs (n=5 per group) underwent percutaneus ischemia/reperfusion (60min/72h or 7 days, respectively). Locked nucleic acid-modified anti-sense miR-92a (LNA-92a) were applied either regionally (antegrade or retrograde) with a catheter or systemically (i.v.). LNA-92a significantly (p<0.01) reduced miR-92a expression in the infarct zone after LNA-92a administration regardless of the application venue. However, catheter-based delivery, but not i.v. infusion, of LNA-92a significantly (p<0.05) reduced the infarct size compared to control LNA treated pigs, which correlated with an improved ejection fraction and LVEDP (p<0.05). Histochemistry revealed that LNA-92a increased capillary density, but decreased leukocyte influx and cardiac cell death. Complete loss of miR-92a in mice attenuated the infarct-related myocardial dysfunction to a larger extent than cardiomyocyte-specific miR-92a deletion. In vitro, LNA-92a protected against hypoxia/reoxygenation-induced cardiomyocyte cell death.
Regional LNA-92a delivery reduces miR-92a levels as well as infarct size and post-ischemic loss of function. LNA-92a exerts cell-protective, pro-angiogenic and anti-inflammatory effects. miR-92a inhibition might be a novel therapeutic tool to preserve cardiac function after ischemia.
[show abstract][hide abstract] ABSTRACT: Rationale: The developmental role of the H3K27 demethylases Jmjd3, especially its epigenetic regulation at target genes in response to upstream developmental signaling is unclear. Objective: To determine the role of Jmjd3 during mesoderm and cardiovascular lineage commitment. Methods and Results: Ablation of Jmjd3 in mouse embryonic stem cells does not affect the maintenance of pluripotency and self-renewal, but compromised mesoderm and subsequent endothelial and cardiac differentiation. Jmjd3 reduces H3K27me3 marks at the Brachyury promoter and facilitates the recruitment ofβ-catenin, which is critical for Wnt signal-induced mesoderm differentiation. Conclusions: These data demonstrate that Jmjd3 is required for mesoderm differentiation and cardiovascular lineage commitment.
Circulation Research 07/2013; · 11.86 Impact Factor
[show abstract][hide abstract] ABSTRACT: MicroRNAs are small non-coding RNAs that regulate gene expression. Although all seven members of the miR-17-92a cluster originate from one primary transcript they are differentially expressed suggesting the presence of posttranscriptional regulation. By RNA pulldown and mass spectrometry we identified SND1, a known regulator of edited RNAs, interacting with pre-miR-92a and all mature miR-17-92a members. Hypoxic conditions lead to an elevation of the pri-miR-17-92a transcript and significantly increased levels of the precursors whereas the mature miRs were not significantly changed. SND1 silencing resolved this block in processing and induced an increase in mature miRs. Together, SND1 might be the missing link between hypoxia and the differential regulation of miRNA processing.
[show abstract][hide abstract] ABSTRACT: AIM/HYPOTHESIS: Monocytes/macrophages play important roles in adipose and vascular tissues and can be polarised as inflammatory M1 or anti-inflammatory M2. We sought to analyse monocyte polarisation status in type 2 diabetes, which is characterised by chronic inflammation. METHODS: We enrolled 60 individuals without diabetes and 53 patients with type 2 diabetes. We quantified standard monocyte subsets defined by cluster of differentiation (CD)14 and CD16. In addition, based on the phenotype of polarised macrophages in vitro, we characterised and quantified more definite M1 (CD68(+)CCR2(+)) and M2 (CX3CR1(+)CD206(+)/CD163(+)) monocytes. We also analysed bone marrow (BM) samples and the effects of granulocyte-colony stimulating factor (G-CSF) stimulation in diabetic and control individuals. RESULTS: We found no alterations in standard monocyte subsets (classical, intermediate and non-classical) when comparing groups. For validation of M1 and M2 phenotypes, we observed that M2 were enriched in non-classical monocytes and had lower TNF-α content, higher LDL scavenging and lower transendothelial migratory capacity than M1. Diabetic patients displayed an imbalanced M1/M2 ratio compared with the control group, attributable to a reduction in M2. The M1/M2 ratio was directly correlated with waist circumference and HbA1c and, among diabetic patients, M2 reduction and M1/M2 increase were associated with microangiopathy. A decrease in M2 was also found in the BM from diabetic patients, with a relative M2 excess compared with the bloodstream. BM stimulation with G-CSF mobilised M2 macrophages in diabetic but not in healthy individuals. CONCLUSIONS/INTERPRETATION: We show that type 2 diabetes markedly reduces anti-inflammatory M2 monocytes through a dysregulation in bone-marrow function. This defect may have a negative impact on microangiopathy.
[show abstract][hide abstract] ABSTRACT: INTRODUCTION: Intracoronary infusion of bone marrow-derived progenitor cells (BMC) in patients with chronic ischaemic heart failure (CHF) is associated with improvement in left ventricular ejection fraction (LVEF), reduction of NT-proBNP levels and improved prognosis. However, effects of this therapy on cardiopulmonary exercise capacity have not been investigated separately so far. PATIENTS AND METHODS: One hundred and fifty-four patients with ischaemic heart failure (mean LVEF 40.3 ± 10.9 %, NT-proBNP 1,103 ± 1,436 pg/ml) underwent cardiopulmonary exercise capacity testing (CPX) before and 3 months after intracoronary infusion of autologous BMC. Thirty patients with a potential bias on the CPX course as concomitant coronary intervention, bypass surgery, new onset of arrhythmias or implantation of cardiac resynchronization devices were excluded from further analysis. RESULTS: The remaining 124 patients showed an increase in exercise time and peak workload by 16.8 and 6 %. Peak oxygen uptake and oxygen uptake efficiency slope also improved by 2.9 and 12.9 %, whereas other parameters like peak oxygen pulse and the slope of minute ventilation versus CO2 elimination remained unchanged. Analysis of patients with poor, moderate and conserved CPX results prior to cell therapy documented that patients in tertiles with lowest initial exercise capacity showed the largest improvements in CPX after therapy. The differences in response to cell therapy were detectable in all investigated CPX parameters and became significant for exercise time, peak oxygen uptake and peak oxygen pulse. These findings indicate that intracoronary BMC therapy improves exercise capacity in CHF patients with more advanced heart failure.
Clinical Research in Cardiology 04/2013; · 3.67 Impact Factor
[show abstract][hide abstract] ABSTRACT: IMPORTANCE The modest effects of clinical studies using intracoronary administration of autologous bone marrow-derived mononuclear cells (BMCs) in patients with chronic postinfarction heart failure may be attributed to impaired homing of BMCs to the target area. Extracorporeal shock wave treatment has been experimentally shown to increase homing factors in the target tissue, resulting in enhanced retention of applied BMCs. OBJECTIVE To test the hypothesis that targeted cardiac shock wave pretreatment with subsequent application of BMCs improves recovery of left ventricular ejection fraction (LVEF) in patients with chronic heart failure. DESIGN, SETTING, AND PARTICIPANTS The CELLWAVE double-blind, randomized, placebo-controlled trial conducted among patients with chronic heart failure treated at Goethe University Frankfurt, Germany, between 2006 and 2011. INTERVENTIONS Single-blind low-dose (n = 42), high-dose (n = 40), or placebo (n = 21) shock wave pretreatment targeted to the left ventricular anterior wall. Twenty-four hours later, patients receiving shock wave pretreatment were randomized to receive double-blind intracoronary infusion of BMCs or placebo, and patients receiving placebo shock wave received intracoronary infusion of BMCs. MAIN OUTCOMES AND MEASURES Primary end point was change in LVEF from baseline to 4 months in the pooled groups shock wave + placebo infusion vs shock wave + BMCs; secondary end points included regional left ventricular function assessed by magnetic resonance imaging and clinical events. RESULTS The primary end point was significantly improved in the shock wave + BMCs group (absolute change in LVEF, 3.2% [95% CI, 2.0% to 4.4%]), compared with the shock wave + placebo infusion group (1.0% [95% CI, -0.3% to 2.2%]) (P = .02). Regional wall thickening improved significantly in the shock wave + BMCs group (3.6% [95% CI, 2.0% to 5.2%]) but not in the shock wave + placebo infusion group (0.5% [95% CI, -1.2% to 2.1%]) (P = .01). Overall occurrence of major adverse cardiac events was significantly less frequent in the shock wave + BMCs group (n = 32 events) compared with the placebo shock wave + BMCs (n = 18) and shock wave + placebo infusion (n = 61) groups (hazard ratio, 0.58 [95% CI, 0.40-0.85]; P = .02). CONCLUSIONS AND RELEVANCE Among patients with postinfarction chronic heart failure, shock wave-facilitated intracoronary administration of BMCs vs shock wave treatment alone resulted in a significant, albeit modest, improvement in LVEF at 4 months. Determining whether the increase in contractile function will translate into improved clinical outcomes requires confirmation in larger clinical end point trials. TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT00326989.
JAMA The Journal of the American Medical Association 04/2013; 309(15):1622-1631. · 29.98 Impact Factor
[show abstract][hide abstract] ABSTRACT: OBJECTIVE: MicroRNAs are important intracellular regulators of gene expression, but also circulate in the blood being protected by extracellular vesicles, proteins, or high-density lipoprotein (HDL). Here, we evaluate the regulation and potential function of HDL- and low-density lipoprotein-bound miRs isolated from healthy subjects and patients with coronary artery disease.Approach and Results-HDL-bound miRs with known effects in the cardiovascular system were analyzed in HDL isolated from healthy subjects (n=10), patients with stable coronary artery disease (n=10), and patients with an acute coronary syndrome (n=10). In HDL from healthy subjects, miR-223 was detected at concentrations >10 000 copies/µg HDL, and miR-126 and miR-92a at about 3000 copies/µg HDL. Concentrations of most miRs were substantially higher in HDL as compared with low-density lipoprotein. However, HDL-bound miR-223 contributed to only 8% of the total circulating miRs. The signatures of miRs varied only slightly in HDL derived from patients with coronary artery disease. We did not observe a significant uptake of HDL-bound miRs into endothelial cells, smooth muscle cells, or peripheral blood mononuclear cells. However, patient-derived HDL transiently reduced miR expression particularly when incubated with smooth muscle and peripheral blood mononuclear cells. CONCLUSIONS: Circulating miRs are detected in HDL and to a lesser extent in low-density lipoprotein, and the miR-signatures are only slightly altered in patients with coronary artery disease. Lipoprotein-bound miRs were not efficiently delivered to endothelial, smooth muscle, and peripheral blood mononuclear cells suggesting that the lipoprotein-associated pool of miRs is not regulating the function of the studied cells in vitro.
Arteriosclerosis Thrombosis and Vascular Biology 04/2013; · 6.34 Impact Factor
[show abstract][hide abstract] ABSTRACT: Ageing is the predominant risk factor for cardiovascular diseases and contributes to a significantly worse outcome in patients with acute myocardial infarction. MicroRNAs (miRNAs) have emerged as crucial regulators of cardiovascular function and some miRNAs have key roles in ageing. We propose that altered expression of miRNAs in the heart during ageing contributes to the age-dependent decline in cardiac function. Here we show that miR-34a is induced in the ageing heart and that in vivo silencing or genetic deletion of miR-34a reduces age-associated cardiomyocyte cell death. Moreover, miR-34a inhibition reduces cell death and fibrosis following acute myocardial infarction and improves recovery of myocardial function. Mechanistically, we identified PNUTS (also known as PPP1R10) as a novel direct miR-34a target, which reduces telomere shortening, DNA damage responses and cardiomyocyte apoptosis, and improves functional recovery after acute myocardial infarction. Together, these results identify age-induced expression of miR-34a and inhibition of its target PNUTS as a key mechanism that regulates cardiac contractile function during ageing and after acute myocardial infarction, by inducing DNA damage responses and telomere attrition.
[show abstract][hide abstract] ABSTRACT: Angiogenesis, defined as blood vessel formation from a preexisting vasculature, is governed by multiple signal cascades including integrin receptors, in particular integrin α(v)β(3). Here we identify the endothelial cell (EC)-secreted factor epidermal growth factor-like protein 7 (EGFL7) as a novel specific ligand of integrin α(v)β(3) thus providing mechanistic insight into its proangiogenic actions in vitro and in vivo. Specifically, EGFL7 attaches to the ECM and by its interaction with integrin α(v)β(3) increases the motility of EC, which allows EC to move on a sticky underground during vessel remodeling. We provide evidence that the deregulation of EGFL7 in zebrafish embryos leads to a severe integrin-dependent malformation of the caudal venous plexus (CVP), pointing towards the significance of EGFL7 in vessel development. In biopsies of patients with neurological diseases, vascular EGFL7 expression rose with increasing EC proliferation. Further, EGFL7 became upregulated in vessels of the stroke penumbra using a mouse model of reversible middle cerebral artery occlusion (MCAO). Our data suggest that EGFL7 expression depends on the remodeling state of the existing vasculature rather than on the phenotype of neurological disease analyzed. In sum, our work sheds a novel light on the molecular mechanism EGFL7 engages to govern physiological and pathological angiogenesis.
[show abstract][hide abstract] ABSTRACT: BACKGROUND: CELL-BASED THERAPIES ARE A PROMISING OPTION IN PATIENTS WITH ACUTE MYOCARDIAL INFARCTION OR CHRONIC HEART FAILURE (CHF). HOWEVER, ADMINISTRATION OF CELLS REQUIRES INTRACORONARY OR INTRACARDIAC INSTRUMENTATION, WHICH IS POTENTIALLY ASSOCIATED WITH PERIPROCEDURAL RISKS. THEREFORE, WE ANALYZED PERIPROCEDURAL COMPLICATIONS AND 30-DAY OUTCOME IN 775 CONSECUTIVE PROCEDURES OF INTRACORONARY ADMINISTRATION OF PROGENITOR CELLS USING THE STOP-FLOW TECHNIQUE.METHODS AND RESULTS: INDICATIONS FOR CELL ADMINISTRATION WERE ACUTE MYOCARDIAL INFARCTION (N=126) AND CHF OF ISCHEMIC (N=562) OR NONISCHEMIC (N=87) ETIOLOGY. VESSEL INJURY WAS OBSERVED IN A TOTAL OF 9 PROCEDURES (1.2%) AND COULD BE PROMPTLY MANAGED BY ADDITIONAL PROGENITOR CELL INJECTION (PCI) IN ALL BUT 1 CASE. NO PROCEDURAL DEATHS WERE OBSERVED. A PERIPROCEDURAL INCREASE IN TROPONIN T WAS OBSERVED IN 3.2% OF THE CHF PROCEDURES, IN WHICH NO CONCOMITANT PCI WAS PERFORMED AND TROPONIN LEVELS WERE NOT ELEVATED BEFORE THE PROCEDURE. INDEPENDENT SIGNIFICANT PREDICTORS OF TROPONIN T INCREASE WERE HIGHER NEW YORK HEART ASSOCIATION (NYHA) CLASS (NYHA I VERSUS NYHA IV; P=0.01; NYHA I VERSUS III; P=0.19; NYHA I VERSUS II; P=0.55), CONCOMITANT REVASCULARIZATION (P0.01), PRESENCE OF ELEVATED TROPONIN T BEFORE THE PROCEDURE (P0.01), AND PERIPHERAL OCCLUSIVE DISEASE (P=0.04). AT 30 DAYS, THERE WERE 4 DEATHS (0.5%), 1 STROKE (0.13%), 8 ACUTE MYOCARDIAL INFARCTIONS (1%), AND 5 HOSPITALIZATIONS FOR EXACERBATION OF HEART FAILURE (0.64%).CONCLUSIONS: INTRACORONARY INFUSION OF PROGENITOR CELLS CAN BE PERFORMED WITH ADEQUATE SAFETY IN PATIENTS WITH ACUTE MYOCARDIAL INFARCTION OR CHF, BECAUSE THE SAFETY PROFILE WAS SIMILAR TO WHAT IS USUALLY EXPECTED FROM A CORONARY ANGIOGRAM IN THE PRESENT COHORT.CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier: NCT00962364, NCT00284713, and NCT00289822.
[show abstract][hide abstract] ABSTRACT: Aging is a complex process that is linked to an increased incidence of major diseases such as cardiovascular and neurodegenerative disease, but also cancer and immune disorders. MicroRNAs (miRNAs) are small non-coding RNAs, which post-transcriptionally control gene expression by inhibiting translation or inducing degradation of targeted mRNAs. MiRNAs target up to hundreds of mRNAs, thereby modulating gene expression patterns. Many miRNAs appear to be dysregulated during cellular senescence, aging and disease. However, only few miRNAs have been so far linked to age-related changes in cellular and organ functions. The present article will discuss these findings, specifically focusing on the cardiovascular and neurological systems.
EMBO Molecular Medicine 01/2013; · 7.80 Impact Factor
[show abstract][hide abstract] ABSTRACT: OBJECTIVE: Histone deacetylases (HDACs) modulate gene expression by deacetylation of histone and nonhistone proteins. Several HDACs control angiogenesis, but the role of HDAC9 is unclear. METHODS AND RESULTS: Here, we analyzed the function of HDAC9 in angiogenesis and its involvement in regulating microRNAs. In vitro, silencing of HDAC9 reduces endothelial cell tube formation and sprouting. Furthermore, HDAC9 silencing decreases vessel formation in a spheroid-based Matrigel plug assay in mice and disturbs vascular patterning in zebrafish embryos. Genetic deletion of HDAC9 reduces retinal vessel outgrowth and impairs blood flow recovery after hindlimb ischemia. Consistently, overexpression of HDAC9 increases endothelial cell sprouting, whereas mutant constructs lacking the catalytic domain, the nuclear localization sequence, or sumoylation site show no effect. To determine the mechanism underlying the proangiogenic effect of HDAC9, we measured the expression of the microRNA (miR)-17-92 cluster, which is known for its antiangiogenic activity. We demonstrate that silencing of HDAC9 in endothelial cells increases the expression of miR-17-92. Inhibition of miR-17-20a rescues the sprouting defects induced by HDAC9 silencing in vitro and blocking miR-17 expression partially reverses the disturbed vascular patterning of HDAC9 knockdown in zebrafish embryos. CONCLUSIONS: We found that HDAC9 promotes angiogenesis and transcriptionally represses the miR-17-92 cluster.
Arteriosclerosis Thrombosis and Vascular Biology 01/2013; · 6.34 Impact Factor
[show abstract][hide abstract] ABSTRACT: Bone marrow (BM) derived stem and progenitor cells contribute to cardiovascular homeostasis and are affected by cardiovascular risk factors. We devised a clinical data-driven approach to test candidate stem cell mobilizing mechanisms in pre-clinical models. We found that PB and BM CD34+ cell counts were directly correlated, and that most circulating CD34+ cells were viable, non-proliferating and derived from the BM. Thus, we analyzed PB and BM CD34+ cell levels as a two-compartment model in 72 patients with or without cardiovascular disease. Self-organizing maps showed that disturbed compartmentalization of CD34+ cells was associated with aging and cardiovascular risk factors especially diabetes. High activity of DPP-4, a regulator of the mobilizing chemokine SDF-1α, was associated with altered stem cell compartmentalization. For validation of these findings, we assessed the role of DPP-4 in the BM mobilization response of diabetic rats. Diabetes differentially affected DPP-4 activity in PB and BM and impaired stem/progenitor cell mobilization after ischemia or G-CSF administration. DPP-4 activity in the BM was required for the mobilizing effect of G-CSF, while in PB it blunted ischemia-induced mobilization. Indeed, DPP-4 deficiency restored ischemia (but not G-CSF)-induced stem cell mobilization and improved vascular recovery in diabetic animals. In conclusion, the analysis of stem cell compartmentalization in humans led us to discover mechanisms of BM unresponsiveness in diabetes determined by tissue-specific DPP-4 dysregulation.
Archiv für Kreislaufforschung 01/2013; 108(1):313. · 7.35 Impact Factor
[show abstract][hide abstract] ABSTRACT: AIMS: Ageing of the immune system, immunosenescence, is characterized by impaired lymphopoiesis, especially B-lymphocyte maturation, and is a hallmark of chronic heart failure (CHF). MicroRNAs (miRNAs) are non-coding, small RNAs, which post-transcriptionally control gene expression of multiple target genes. The miR-181 family is known to control haematopoietic lineage differentiation. Here, we study the role of the miR-181 family in immunosenescence and CHF. METHODS AND RESULTS: We conducted a clinical study analysing peripheral blood (PB) for miRNA expression and leucocyte distribution of young healthy controls (25 ± 4 years; n = 30), aged healthy controls (64 ± 5 years; n = 13), and age-matched CHF patients (64 ± 11years; n = 18). The expression of miR-181 family members was reduced, whereas miR-34a was increased in PB of aged individuals. In particular, miR-181c was further reduced in age-matched CHF patients. In PB, we observed reduced numbers of lymphocytes, in particular cytotoxic T cells and B cells, with rising age, and the expression of miR-181 correlated with the number of B cells. Notably, in CHF patients, ischaemic heart failure was associated with a further reduction of total B cells as well as their subpopulations, such as memory B cells, compared with age-matched healthy volunteers. CONCLUSIONS: Ageing- and CHF-associated changes in PB leucocyte subsets are paralleled by alterations in the expression of miRNAs involved in lymphopoiesis, which might play an important role in the age-related and CHF-mediated dysregulation of immune functions resulting in immunosenescence. Furthermore, miR-181c may serve as a marker for reduced immune functions in CHF patients.
European Journal of Heart Failure 12/2012; · 5.25 Impact Factor
[show abstract][hide abstract] ABSTRACT: RATIONALE: MicroRNAs (miRNAs or miR) are implicated in the pathogenesis of various cardiovascular diseases, including pulmonary arterial hypertension (PAH). OBJECTIVE: We sought to measure changes in plasma levels of miRNAs in PAH patients and relate them to the severity of the disease. METHODS: A microarray screen was performed on total plasma RNA from 8 patients with PAH and 8 healthy controls. Quantitative PCR confirmed reduced miR-150 concentrations and was then used to measure miR-150 levels in (a) two separate cohorts of PAH patients, from London (n=145) and Sheffield (n=30), respectively; (b) circulating microvesicles and blood cells; and (c) lungs from a monocrotaline rat model. MEASUREMENTS AND MAIN RESULTS: 58 miRNAs showed differences in plasma concentration and miR-150 the largest down-regulation in PAH. Receiver-operator-characteristic analysis showed both raw and normalized plasma miR-150 levels correlated with 2-year survival (P<0.01) in PAH patients. Cox regression analysis confirmed miR-150 levels as a significant predictor of survival. Age, baseline cardiac index, WHO functional class, 6 minute walk distance, disease duration and red cell distribution width also predicted survival. Entering these covariates in a multivariable model verified plasma miR-150 levels as an independent predictor of survival in PAH (HR: 0.533, P=0.010). miR-150 levels also predicted survival in a second, independent PAH cohort. miR-150 levels were significantly reduced in circulating microvesicles from PAH patients and the lungs of the monocrotaline rat. CONCLUSIONS: Reduced circulating miR-150 levels are associated with poor survival in PAH.
American Journal of Respiratory and Critical Care Medicine 12/2012; · 11.04 Impact Factor
[show abstract][hide abstract] ABSTRACT: Background: Prognosis of patients with heart failure remains poor despite improved conventional and interventional treatment regimens. The improvement of neovascularisation and repair processes by administration of bone marrow-derived cells modestly improved the recovery after acute myocardial infarction. However, circulating patient-derived cells are reduced in number and function particularly in chronic heart failure. Therefore, we tested the hypothesis whether the mobilisation of circulating mononuclear proangiogenic cells (CPCs) by G-CSF may overcome some of these limitations.Methods: In the present pilot study, 32 patients with at least 3 months old myocardial infarction were randomized to G-CSF alone (G-CSF group) or intracoronary infusion of GCSF-mobilised and cultured CPCs into the infarct-related artery (G-CSF/CPC group). Primary endpoint of the study was safety. Efficacy parameters included serial assessment of LV function, NT-proBNP levels and cardiopulmonary exercise testing.Results: G-CSF effectively mobilised circulating CD34⁺CD45⁺ cells after 5 days in all patients (408 ± 64%) without serious adverse events.At 3 months, NYHA class and global LV function did not show significant improvements in both treatment groups (G-CSF: Δ LVEF 1.6 ± 2.4 %; p = 0.10; G-CSF/CPC: Δ LVEF 1.4 ± 4.1%; p = 0.16). In contrast, target area contractility improved significantly in the G-CSF/CPC group. During 5-year follow-up, 1 patient died after rehospitalisation for worsening heart failure. 11 patients underwent further revascularization procedures. NT-proBNP levels, cardiopulmonary exercise capacity and NYHA class remained stable in both treatment groups.Conclusion: The results from our pilot trial indicate that administration of G-CSF alone or of G-CSF-mobilised and cultured CPCs can be performed safely in patients with chronic ischemic heart disease. However, only minor effects on LV-function, NT-proBNP levels and NYHA classification were observed during follow-up suggesting that the enhancement of CPCs by G-CSF alone does not substantially improve intracoronary cell therapy effects in patients with chronic ischemic heart failure.