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ABSTRACT: It is well known that altered blood flow is related to vascular diseases, including atherosclerosis, restenosis and arteriosclerosis, which preferentially located at areas with disturbed blood flow, suggesting that altered biomechanical stress may exert their effect on the vascular disease. Recent evidence indicated the presence of abundant stem/progenitor cells in the vessel wall, in which laminar shear stress can stimulate these cells to differentiate toward endothelial lineage, while cyclic strain results in smooth muscle differentiation. In line with this, it was evidenced that altered biomechanical stress in stented vessels may lead to "wrong" direction of vascular stem cell differentiation resulting in restenosis. However, the underlying mechanisms are not well understood. In this article, we will give an overview of the effect of local flow pattern on stem/progenitor cell differentiation and the possible mechanism on how blood flow influences stem cell behaviours in the development of vascular diseases.
Cardiovascular research 03/2013; · 5.80 Impact Factor
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ABSTRACT: OBJECTIVE: Diabetes mellitus causes bone marrow (BM) microangiopathy. This study aimed to investigate the mechanisms responsible for BM endothelial dysfunction in diabetes mellitus. METHODS AND RESULTS: The analysis of differentially expressed transcripts in BM endothelial cells (BMECs) from type-1 diabetic and nondiabetic mice showed an effect of diabetes mellitus on signaling pathways controlling cell death, migration, and cytoskeletal rearrangement. Type-1 diabetic-BMECs displayed high reactive oxygen species levels, increased expression and activity of RhoA and its associated protein kinases Rho-associated kinase 1/Rho-associated kinase 2, and reduced Akt phosphorylation/activity. Likewise, diabetes mellitus impaired Akt-related BMEC functions, such as migration, network formation, and angiocrine factor-releasing activity, and increased vascular permeability. Moreover, high glucose disrupted BMEC contacts through Src tyrosine kinase phosphorylation of vascular endothelial cadherin. These alterations were prevented by constitutively active Akt (myristoylated Akt), Rho-associated kinase inhibitor Y-27632, and Src inhibitors. Insulin replacement restored BMEC abundance, as assessed by flow cytometry analysis of the endothelial marker MECA32, and endothelial barrier function in BM of type-1 diabetic mice. CONCLUSIONS: Redox-dependent activation of RhoA/Rho-associated kinase and Src/vascular endothelial cadherin signaling pathways, together with Akt inactivation, contribute to endothelial dysfunction in diabetic BM. Metabolic control is crucial for maintenance of EC homeostasis and endothelial barrier function in BM of diabetic mice.
Arteriosclerosis Thrombosis and Vascular Biology 01/2013; · 6.37 Impact Factor
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Circulation Research 01/2013; 112(1):3-5. · 9.49 Impact Factor
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Gaia Spinetti,
Daniela Cordella,
Orazio Fortunato,
Elena Sangalli,
Sergio P Losa,
Ambra Gotti,
Franco Carnelli,
Francesco Rosa,
Stefano Riboldi,
Fausto Sessa,
Elisa Avolio,
Antonio Paolo Beltrami, Costanza Emanueli,
Paolo R Madeddu
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ABSTRACT: Rationale: The impact of diabetes on bone marrow (BM) structure is incompletely understood. Objective: Investigate the effect of type-2 diabetes (T2D) on BM microvascular and hematopoietic cell composition in patients with/out vascular complications. Methods and Results: Bone samples were obtained from T2D patients and non-diabetic controls (C) during hip replacement surgery and from T2D patients undergoing amputation for critical limb ischemia (CLI). BM composition was assessed by histomorphometry, immunostaining and flow cytometry. Expressional studies were performed on CD34(pos) immuno-sorted BM progenitor cells (PCs). Diabetes causes a reduction of hematopoietic tissue, fat deposition, and microvascular rarefaction, especially when associated to CLI. Immunohistochemistry documented increased apoptosis and reduced abundance of CD34(pos)-PCs in diabetic groups. Likewise, flow cytometry showed scarcity of BM PCs in T2D and T2D+CLI compared to C, but similar levels of mature hematopoietic cells. Activation of apoptosis in CD34(pos-)PCs was associated to upregulation and nuclear localization of the pro-apoptotic factor FOXO3a and induction of FOXO3a targets, p21 and p27(kip1). Moreover, miR-155, which regulates cell survival through inhibition of FOXO3a, was downregulated in diabetic CD34(pos)-PCs and inversely correlated with FOXO3a levels. The effect of diabetes on anatomical and molecular endpoints was confirmed when considering background covariates. Furthermore, exposure of healthy CD34(pos)-PCs to high glucose reproduced the transcriptional changes induced by diabetes, with this effect being reversed by forced expression of miR-155. Conclusions: We provide new anatomical and molecular evidence for the damaging effect of diabetes on human BM, comprising microvascular rarefaction and shortage of PCs due to activation of pro-apoptotic pathway.
Circulation Research 12/2012; · 9.49 Impact Factor
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Gaia Spinetti,
Orazio Fortunato,
Andrea Caporali,
Saran Shantikumar,
Micol Marchetti,
Marco Meloni,
Ilaria Floris,
Betty Descamps,
Elena Sangalli,
Rosa Vono,
Ezio Faglia,
Claudia Specchia,
Gianfranco Pintus,
Paolo R Madeddu, Costanza Emanueli
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ABSTRACT: Rationale: Circulating Proangiogenic Cells (PACs) support post-ischemic neovascularization. Cardiovascular disease and diabetes impair PAC regenerative capacities via not fully known molecular mechanisms. We hypothesize a role for microRNAs (miRs). Circulating miRs are currently investigated as potential diagnostic and prognostic biomarkers. Objective: 1) To profile miR expression in PACs from critical limb ischemia (CLI) patients; 2) To demonstrate that miR-15a and miR-16 regulate PAC functions; 3) To characterize circulating miR-15a and miR-16 and to investigate their potential biomarker value. Methods and Results: Twenty-eight miRs potentially able to modulate angiogenesis were measured in PACs from CLI patients with/out diabetes and controls. miR-15a and miR-16 were further analyzed. CLI-PACs expressed higher level of mature miR-15a and miR-16 and of the primary transcript primiR-15a/16-1. miR-15a/-16 overexpression impaired healthy PACs survival and migration. Conversely, miR-15a/-16 inhibition improved CLI-PAC defective migration. VEGF-A and AKT-3 were validated as direct targets of the two miRs and their protein levels were reduced in miR-15a/-16-overexpressing healthy PACs and in CLI-PACs. Transplantation of healthy PACs ex-vivo engineered with anti-miR-15a/-16 improved post-ischemic blood flow recovery and muscular arteriole density in immunodeficient mice. miR-15a and miR-16 were present in human blood, including conjugated to Argonaute-2 and in exosomes. Both miRs were increased in the serum of CLI patients and positively correlated with amputation after restenosis at 12 months post-revascularization of CLI-T2D patients. Serum miR-15a additionally correlated with restenosis at follow-up. Conclusions: 1) Ex-vivo miR-15a/16 inhibition enhances PAC therapeutic potential; 2) circulating miR-15a deserves further investigation as prognostic biomarker in CLI patients undergoing revascularization.
Circulation Research 12/2012; · 9.49 Impact Factor
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Andrea Caporali,
Marco Meloni,
Ashley M Miller,
Klemens Vierlinger,
Alessandro Cardinali,
Gaia Spinetti,
Audrey Nailor,
Ezio Faglia,
Sergio Losa,
Ambra Gotti,
Orazio Fortunato,
Tijana Mitic,
Manuela Hofner,
Christa Noehammer,
Paolo Madeddu, Costanza Emanueli
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ABSTRACT: OBJECTIVE: The p75 neurotrophin receptor (p75(NTR)) contributes to diabetes mellitus-induced defective postischemic neovascularization. The interleukin-33 receptor ST2 is expressed as transmembrane (ST2L) and soluble (sST2) isoforms. Here, we studied the following: (1) the impact of p75(NTR) in the healing of ischemic and diabetic calf wounds; (2) the link between p75(NTR) and ST2; and (3) circulating sST2 levels in critical limb ischemia (CLI) patients. METHODS AND RESULTS: Diabetes mellitus was induced in p75(NTR) knockout (p75KO) mice and wild-type (WT) littermates by streptozotocin. Diabetic and nondiabetic p75KO and WT mice received left limb ischemia induction and a full-thickness wound on the ipsilateral calf. Diabetes mellitus impaired wound closure and angiogenesis and increased ST2 expression in WT, but not in p75KO wounds. In cultured endothelial cells, p75(NTR) promoted ST2 (both isoforms) expression through p38(MAPK)/activating transcription factor 2 pathway activation. Next, sST2 was measured in the serum of patients with CLI undergoing either revascularization or limb amputation and in the 2 nondiabetic groups (with CLI or nonischemic individuals). Serum sST2 increased in diabetic patients with CLI and was directly associated with higher mortality at 1 year from revascularization. CONCLUSIONS: p75(NTR) inhibits the healing of ischemic lower limb wounds in diabetes mellitus and promotes ST2 expression. Circulating sST2 predicts mortality in diabetic CLI patients.
Arteriosclerosis Thrombosis and Vascular Biology 10/2012; · 6.37 Impact Factor
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ABSTRACT: AIMS: Diabetes impinges upon mechanisms of cardiovascular repair. However, the biochemical adaptation of cardiac stem cells to sustained hyperglycaemia remains largely unknown. Here, we investigate the molecular targets of high glucose-induced damage in cardiac progenitor cells (CPCs) from murine and human hearts and attempt safeguarding CPC viability and function through reactivation of the pentose phosphate pathway. METHODS AND RESULTS: Type-1 diabetes was induced by streptozotocin. CPC abundance was determined by flow cytometry. Proliferating CPCs were identified in situ by immunostaining for the proliferation marker Ki67. Diabetic hearts showed marked reduction in CPC abundance and proliferation when compared with controls. Moreover, Sca-1(pos) CPCs isolated from hearts of diabetic mice displayed reduced activity of key enzymes of the pentose phosphate pathway, glucose-6-phosphate dehydrogenase (G6PD), and transketolase, increased levels of superoxide and advanced glucose end-products (AGE), and inhibition of the Akt/Pim-1/Bcl-2 signalling pathway. Similarly, culture of murine CPCs or human CD105(pos) progenitor cells in high glucose inhibits the pentose phosphate and pro-survival signalling pathways, leading to the activation of apoptosis. In vivo and in vitro supplementation with benfotiamine reactivates the pentose phosphate pathway and rescues CPC availability and function. This benefit is abrogated by either G6PD silencing by small interfering RNA (siRNA) or Akt inhibition by dominant-negative Akt. CONCLUSION: We provide new evidence of the negative impact of diabetes and high glucose on mechanisms controlling CPC redox state and survival. Boosting the pentose phosphate pathway might represent a novel mechanistic target for protection of CPC integrity.
Cardiovascular research 09/2012; · 5.80 Impact Factor
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ABSTRACT: Stem cell (SC) therapy represents a big hope for treating unmet clinical needs, including in the context of cardiovascular disease. The abilities of embryonic SC (ESCs) to self-renew indefinitely and to differentiate in all the three germ layers make these SCs very attractive for both basic science investigations and clinical therapies. ESCs can generate vascular endothelial and mural cells to be used for transplantation and to create engineered organs. Moreover ESC can be used to mimic developmental vasculogenesis and angiogenesis in vitro. However, additional studies are needed to improve vascular differentiation protocols of ESCs. This review focuses on ESCs and the technologies allowing for their differentiation into mesoderm and vascular lineage. Moreover, the potential of ESC-derived vascular cells for clinical cardiovascular therapies and strategies to improve ESC engraftment efficiency are discussed.
Vascular Pharmacology 04/2012; 56(5-6):267-79. · 1.99 Impact Factor
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Silvia Amadesi,
Carlotta Reni,
Rajesh Katare,
Marco Meloni,
Atsuhiko Oikawa,
Antonio P Beltrami,
Elisa Avolio,
Daniela Cesselli,
Orazio Fortunato,
Gaia Spinetti,
Raimondo Ascione,
Elisa Cangiano,
Marco Valgimigli,
Stephen P Hunt, Costanza Emanueli,
Paolo Madeddu
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ABSTRACT: Pain triggers a homeostatic alarm reaction to injury. It remains unknown, however, whether nociceptive signaling activated by ischemia is relevant for progenitor cells (PC) release from bone marrow. To this end, we investigated the role of the neuropeptide substance P (SP) and cognate neurokinin 1 (NK1) nociceptor in PC activation and angiogenesis during ischemia in mice and in human subjects.
The mouse bone marrow contains sensory fibers and PC that express SP. Moreover, SP-induced migration provides enrichment for PC that express NK1 and promote reparative angiogenesis after transplantation in a mouse model of limb ischemia. Acute myocardial infarction and limb ischemia increase SP levels in peripheral blood, decrease SP levels in bone marrow, and stimulate the mobilization of NK1-expressing PC, with these effects being abrogated by systemic administration of the opioid receptor agonist morphine. Moreover, bone marrow reconstitution with NK1-knockout cells results in depressed PC mobilization, delayed blood flow recovery, and reduced neovascularization after ischemia. We next asked whether SP is instrumental to PC mobilization and homing in patients with ischemia. Human PC express NK1, and SP-induced migration provides enrichment for proangiogenic PC. Patients with acute myocardial infarction show high circulating levels of SP and NK1-positive cells that coexpress PC antigens, such as CD34, KDR, and CXCR4. Moreover, NK1-expressing PC are abundant in infarcted hearts but not in hearts that developed an infarct after transplantation.
Our data highlight the role of SP in reparative neovascularization. Nociceptive signaling may represent a novel target of regenerative medicine.
Circulation 03/2012; 125(14):1774-86, S1-19. · 14.74 Impact Factor
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Nicole M Kane,
Lynsey Howard,
Betty Descamps,
Marco Meloni,
John McClure,
Ruifang Lu,
Angela McCahill,
Christopher Breen,
Ruth M Mackenzie,
Christian Delles,
Joanne C Mountford,
Graeme Milligan, Costanza Emanueli,
Andrew H Baker
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ABSTRACT: MicroRNAs (miRNAs) are short noncoding RNAs, which post-transcriptionally regulate gene expression. miRNAs are transcribed as precursors and matured to active forms by a series of enzymes, including Dicer. miRNAs are important in governing cell differentiation, development, and disease. We have recently developed a feeder- and serum-free protocol for direct derivation of endothelial cells (ECs) from human embryonic stem cells (hESCs) and provided evidence of increases in angiogenesis-associated miRNAs (miR-126 and -210) during the process. However, the functional role of miRNAs in hESC differentiation to vascular EC remains to be fully interrogated. Here, we show that the reduction of miRNA maturation induced by Dicer knockdown suppressed hES-EC differentiation. A miRNA microarray was performed to quantify hES-EC miRNA profiles during defined stages of endothelial differentiation. miR-99b, -181a, and -181b were identified as increasing in a time- and differentiation-dependent manner to peak in mature hESC-ECs and adult ECs. Augmentation of miR-99b, -181a, and -181b levels by lentiviral-mediated transfer potentiated the mRNA and protein expression of EC-specific markers, Pecam1 and VE Cadherin, increased nitric oxide production, and improved hES-EC-induced therapeutic neovascularization in vivo. Conversely, knockdown did not impact endothelial differentiation. Our results suggest that miR-99b, -181a, and -181b comprise a component of an endothelial-miRNA signature and are capable of potentiating EC differentiation from pluripotent hESCs.
Stem Cells 01/2012; 30(4):643-54. · 7.78 Impact Factor
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ABSTRACT: The term angiogenesis describes the growth of endothelial sprouts from preexisting postcapillary venules. More recently, this term has been used to generally indicate the growth and remodeling process of the primitive vascular network into a complex network during development. In adulthood, angiogenesis is activated as a reparative process during wound healing and following ischemia, and it plays a key role in tumor growth and metastasis as well as in inflammatory diseases and diabetic retinopathy. MicroRNAs (miRNAs) are endogenous, small, noncoding RNAs that negatively control gene expression of target mRNAs. In this paper, we aim at describing the role of miRNAs in postischemic angiogenesis. First, we will describe the regulation and the expression of miRNAs in endothelial cells. Then, we will analyze the role of miRNAs in postischemic vascular repair. Finally, we will discuss the role of circulating miRNAs as potential biomarkers in ischemic diseases.
Cardiology research and practice. 01/2012; 2012:486702.
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ABSTRACT: Diabetes is a cause of cardiac dysfunction, reduced myocardial perfusion, and ultimately heart failure. Nerve growth factor (NGF) exerts protective effects on the cardiovascular system. This study investigated whether NGF gene transfer can prevent diabetic cardiomyopathy in mice. We worked with mice with streptozotocin-induced type 1 diabetes and with nondiabetic control mice. After having established that diabetes reduces cardiac NGF mRNA expression, we tested NGF gene therapies with adeno-associated viral vectors (AAVs) for the capacity to protect the diabetic mouse heart. To this aim, after 2 weeks of diabetes, cardiac expression of human NGF or β-Gal (control) genes was induced by either intramyocardial injection of AAV serotype 2 (AAV2) or systemic delivery of AAV serotype 9 (AAV9). Nondiabetic mice were given AAV2-β-Gal or AAV9-β-Gal. We found that the diabetic mice receiving NGF gene transfer via either AAV2 or AAV9 were spared the progressive deterioration of cardiac function and left ventricular chamber dilatation observed in β-Gal-injected diabetic mice. Moreover, they were additionally protected from myocardial microvascular rarefaction, hypoperfusion, increased deposition of interstitial fibrosis, and increased apoptosis of endothelial cells and cardiomyocytes, which afflicted the β-Gal-injected diabetic control mice. Our data suggest therapeutic potential of NGF for the prevention of cardiomyopathy in diabetic subjects.
Diabetes 01/2012; 61(1):229-40. · 8.29 Impact Factor
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ABSTRACT: Diabetes is the most common metabolic disorder and is recognized as one of the most important health threats of our time. MicroRNAs (miRNAs) are a novel group of non-coding small RNAs that have been implicated in a variety of physiological processes, including glucose homeostasis. Recent research has suggested that miRNAs play a critical role in the pathogenesis of diabetes and its related cardiovascular complications. This review focuses on the aberrant expression of miRNAs in diabetes and examines their role in the pathogenesis of endothelial dysfunction, cardiovascular disease, and diabetic retinopathy. Furthermore, we discuss the potential role of miRNAs as blood biomarkers and examine the potential of therapeutic interventions targeting miRNAs in diabetes.
Cardiovascular research 11/2011; 93(4):583-93. · 5.80 Impact Factor
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Rajesh Katare,
Federica Riu,
Kathryn Mitchell,
Miriam Gubernator,
Paola Campagnolo,
Yuxin Cui,
Orazio Fortunato,
Elisa Avolio,
Daniela Cesselli,
Antonio Paolo Beltrami,
Gianni Angelini, Costanza Emanueli,
Paolo Madeddu
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ABSTRACT: Pericytes are key regulators of vascular maturation, but their value for cardiac repair remains unknown.
We investigated the therapeutic activity and mechanistic targets of saphenous vein-derived pericyte progenitor cells (SVPs) in a mouse myocardial infarction (MI) model.
SVPs have a low immunogenic profile and are resistant to hypoxia/starvation (H/S). Transplantation of SVPs into the peri-infarct zone of immunodeficient CD1/Foxn-1(nu/nu) or immunocompetent CD1 mice attenuated left ventricular dilatation and improved ejection fraction compared to vehicle. Moreover, SVPs reduced myocardial scar, cardiomyocyte apoptosis and interstitial fibrosis, improved myocardial blood flow and neovascularization, and attenuated vascular permeability. SVPs secrete vascular endothelial growth factor A, angiopoietin-1, and chemokines and induce an endogenous angiocrine response by the host, through recruitment of vascular endothelial growth factor B expressing monocytes. The association of donor- and recipient-derived stimuli activates the proangiogenic and prosurvival Akt/eNOS/Bcl-2 signaling pathway. Moreover, microRNA-132 (miR-132) was constitutively expressed and secreted by SVPs and remarkably upregulated, together with its transcriptional activator cyclic AMP response element-binding protein, on stimulation by H/S or vascular endothelial growth factor B. We next investigated if SVP-secreted miR-132 acts as a paracrine activator of cardiac healing. In vitro studies showed that SVP conditioned medium stimulates endothelial tube formation and reduces myofibroblast differentiation, through inhibition of Ras-GTPase activating protein and methyl-CpG-binding protein 2, which are validated miR-132 targets. Furthermore, miR-132 inhibition by antimiR-132 decreased SVP capacity to improve contractility, reparative angiogenesis, and interstitial fibrosis in infarcted hearts.
SVP transplantation produces long-term improvement of cardiac function through a novel paracrine mechanism involving the secretion of miR-132 and inhibition of its target genes.
Circulation Research 08/2011; 109(8):894-906. · 9.49 Impact Factor
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ABSTRACT: MicroRNAs (miRs) are post-transcriptional inhibitory regulators of gene expression acting by direct binding to complementary messenger RNA (mRNA) transcripts. Recent studies have demonstrated that miRs are crucial determinants of endothelial cell behavior and angiogenesis. We have provided evidence of the prominent role of miR-503 in impairment of postischemic reparative angiogenesis in the setting of diabetes. Because miR-503 belongs to the miR-16 extended family of miRs, in this review, we describe the cardiovascular functions of miR-503 and other members of the miR-16 family and their impact on angiogenesis.
Trends in cardiovascular medicine 08/2011; 21(6):162-6. · 4.37 Impact Factor
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ABSTRACT: The term angiogenesis derives from the Greek words 'angeio' meaning blood vessel, and 'genesis' meaning production or birth, together referring to the creation of blood vessels within the body. This term has been used to generally indicate the growth and remodeling process of the primitive vascular network into a complex network during pre-natal development. After birth, reparative angiogenesis is activated during wound healing and in response to ischemia, while pathological angiogenesis contributes to tumor growth and metastasis, arthritis and ocular diseases, such as diabetic retinopathy. MicroRNAs (miRNAs) are a class of endogenous, small, non-coding RNAs that control gene expression by acting on target mRNAs for promoting either their degradation or translational repression. There is increasing evidence that miRNAs play important roles in vascular development as well as in vascular diseases. In this review, we aim at describing the role of miRNAs in angiogenesis, focusing, in particular, on post-ischemic neovascularization. First, we will describe the regulation and the expression of miRNAs in endothelial cells. Then, we will analyze the role of miRNAs in reparative and pathological angiogenesis. Finally, we will discuss the innovative strategies available to inhibit the level of pathogenic anti-angiogenic miRNAs and to increase expression of therapeutic miRNAs.
Vascular Pharmacology 07/2011; 55(4):79-86. · 1.99 Impact Factor
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Rajesh Katare,
Andrea Caporali,
Lorena Zentilin,
Elisa Avolio,
Graciela Sala-Newby,
Atsuhiko Oikawa,
Daniela Cesselli,
Antonio Paolo Beltrami,
Mauro Giacca, Costanza Emanueli,
Paolo Madeddu
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ABSTRACT: Studies in transgenic mice showed the key role of (Pim-1) (proviral integration site for Moloney murine leukemia virus-1) in the control of cardiomyocyte function and viability.
We investigated whether Pim-1 represents a novel mechanistic target for the cure of diabetic cardiomyopathy, a steadily increasing cause of nonischemic heart failure.
In streptozotocin-induced type 1 diabetic mice, Pim-1 protein levels declined during progression of cardiomyopathy, along with upregulation of Pim-1 inhibitors, protein phosphatase 2A, and microRNA-1. Moreover, diabetic hearts showed low levels of antiapoptotic B-cell lymphoma-2 (Bcl-2) protein and increased proapoptotic caspase-3 activity. Studies on adult rat cardiomyocytes and murine cardiac progenitor cells challenged with high glucose confirmed the in vivo expressional changes. In rescue studies, anti-microRNA-1 boosted Pim-1 and Bcl-2 expression and promoted cardiomyocyte and cardiac progenitor cell survival under high glucose conditions. Similarly, transfection with Pim-1 plasmid prevented high glucose-induced cardiomyocyte and cardiac progenitor cell apoptosis. Finally, a single intravenous injection of human PIM-1 via cardiotropic serotype-9 adeno-associated virus (1 × 10(10) or 5 × 10(10) plaque-forming units per animal) at 4 weeks after diabetes induction led to sustained cardiac overexpression of Pim-1 and improved diastolic function and prevented left ventricular dilation and failure. Histological examination showed reduced cardiomyocyte apoptosis and fibrosis in association with increased c-kit(+) cells and cardiomyocyte proliferation, whereas molecular analysis confirmed activation of the prosurvival pathway and conservation of sarcoendoplasmic reticulum Ca(2+)-ATPase and α-myosin heavy chain in Pim-1-treated hearts.
Pim-1 downregulation contributes in the pathogenesis of diabetic cardiomyopathy. Systemic delivery of human PIM-1 via cardiotropic adeno-associated virus serotype-9 represents a novel and effective approach to treat diabetic cardiomyopathy.
Circulation Research 05/2011; 108(10):1238-51. · 9.49 Impact Factor
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Gaia Spinetti,
Orazio Fortunato,
Daniela Cordella,
Paola Portararo,
Nicolle Kränkel,
Rajesh Katare,
Graciela B Sala-Newby,
Christine Richer,
Marie-Pascale Vincent,
Francois Alhenc-Gelas,
Giancarlo Tonolo,
Sara Cherchi, Costanza Emanueli,
Paolo Madeddu
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ABSTRACT: Homing of proangiogenic cells (PACs) is guided by chemoattractants and requires proteases to disrupt the extracellular matrix. The possibility that PAC recruitment involves an interaction between proteases and chemotactic factor receptors remains largely unexplored.
To determine the role of human tissue kallikrein (hK1) in PAC invasion and its dependency on kinin receptor signaling.
Human mononuclear cells (MNCs) and culture-selected PACs express and release mature hK1 protein. HK1 gene (KLK1) silencing reduced PACs migratory, invasive, and proangiogenic activities. KLK1-knockout mouse bone marrow-derived MNCs showed similar impairments and were unable to support reparative angiogenesis in a mouse model of peripheral ischemia. Conversely, adenovirus-mediated KLK1 (Ad.KLK1) gene transfer enhanced PAC-associated functions, whereas the catalytically inactive variant R53H-KLK1 was ineffective. HK1-induced effects are mediated by a kinin B(2) receptor (B(2)R)-dependent mechanism involving inducible nitric oxide synthase and metalloproteinase-2 (MMP2). Lower hK1 protein levels were observed in PACs from type 2 diabetic (T2D) patients, whereas KLK1 mRNA levels were similar to those of healthy subjects, suggesting a post-transcriptional defect. Furthermore, B(2)R is normally expressed on T2D-PACs but remains uncoupled from downstream signaling. Importantly, whereas Ad.KLK1 alone could not restore T2D-PAC invasion capacity, combined KLK1 and B(2)R expression rescued the diabetic phenotype.
This study reveals new interactive components of the PACs invasive machinery, acting via protease- and kinin receptor-dependent mechanisms.
Circulation Research 02/2011; 108(3):284-93. · 9.49 Impact Factor
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Andrea Caporali,
Marco Meloni,
Christine Völlenkle,
Desiree Bonci,
Graciela B Sala-Newby,
Roberta Addis,
Gaia Spinetti,
Sergio Losa,
Rachel Masson,
Andrew H Baker,
Reuven Agami,
Carlos le Sage,
Gianluigi Condorelli,
Paolo Madeddu,
Fabio Martelli, Costanza Emanueli
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ABSTRACT: Diabetes mellitus impairs endothelial cell (EC) function and postischemic reparative neovascularization by molecular mechanisms that are not fully understood. microRNAs negatively regulate the expression of target genes mainly by interaction in their 3' untranslated region.
We found that microRNA-503 (miR-503) expression in ECs is upregulated in culture conditions mimicking diabetes mellitus (high D-glucose) and ischemia-associated starvation (low growth factors). Under normal culture conditions, lentivirus-mediated miR-503-forced expression inhibited EC proliferation, migration, and network formation on Matrigel (comparisons versus lentivirus.GFP control). Conversely, blocking miR-503 activity by either adenovirus-mediated transfer of a miR-503 decoy (Ad.decoymiR-503) or by antimiR-503 (antisense oligonucleotide) improved the functional capacities of ECs cultured under high D-glucose/low growth factors. We identified CCNE1 and cdc25A as direct miR-503 targets which are downregulated by high glucose/low growth factors in ECs. Next, we obtained evidence that miR-503 expression is increased in ischemic limb muscles of streptozotocin-diabetic mice and in ECs enriched from these muscles. Moreover, Ad.decoymiR-503 delivery to the ischemic adductor of diabetic mice corrected diabetes mellitus-induced impairment of postischemic angiogenesis and blood flow recovery. We finally investigated miR-503 and target gene expression in muscular specimens from the amputated ischemic legs of diabetic patients. As controls, calf biopsies of nondiabetic and nonischemic patients undergoing saphenous vein stripping were used. In diabetic muscles, miR-503 expression was remarkably higher, and it inversely correlated with cdc25 protein expression. Plasma miR-503 levels were also elevated in the diabetic individuals.
Our data suggest miR-503 as a possible therapeutic target in diabetic patients with critical limb ischemia.
Circulation 01/2011; 123(3):282-91. · 14.74 Impact Factor
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ABSTRACT: Several types of stem and progenitor cells are currently under investigation for their potential to accomplish vascular regeneration. This review focuses on embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). We will discuss the technologies allowing for their derivation, culture expansion and maintenance in a pluripotent status. Moreover, both ESCs and iPSCs can be differentiated in endothelial cells (ECs) and mural cell, including vascular smooth muscle cells (VSMCs). Here, we will describe the involvements of growth factors (vascular endothelial growth factors-VEGFs-, platet-derived growth factors-PDGFs-), Wnt and Notch signal pathways, reactive oxygen species (ROS), histone deacetylases (HDACs), and microRNAs (miRNAs) in vascular cell differentiation from pluripotent stem cells. We will additionally describe the therapeutic potential of stem cells for vascular medicine.
Pharmacology [?] Therapeutics 10/2010; 129(1):29-49. · 8.56 Impact Factor
