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PubMed1996

The temporal and spatial vascular endothelial growth factor expression in retinal vasculogenesis of rat neonates.

Vascular endothelial growth factor (VEGF) has been identified as an endothelial cell-specific mitogen with potent angiogenic properties. VEGF is overexpressed in pathologic angiogenesis observed in tumor growth, rheumatoid arthritis, and retinal angiogenic diseases such as diabetic retinopathy and retinopathy of prematurity. VEGF expression in physiologic angiogenesis, i.e., vasculogenesis, has also been reported in the embryonic organs such as brain, kidney, spleen, and lung. However, the details of VEGF expression in vasculogenesis remain largely unclear. To determine if VEGF contributes to vasculogenesis in the developing tissues, VEGF expression was studied by both immunohistochemistry and in situ hybridization in newborn rat retinas on postnatal days 3, 7, 14, and 30. Vasculogenesis was assessed by both the ink perfusion method and a histologic examination. To identify the cell types of VEGF-expressing cells, immunohistochemistry for cell markers such as glial fibrillary acidic protein and von Willebrand factor was performed. On postnatal days 3 and 7, when retinal vasculogenesis was active, VEGF mRNA and protein(s) were prominently expressed in the ganglion cell and the inner nuclear layers. In rats, as well as humans, these two layers are where the retinal vessels develop, and these two layers depend solely on the retinal vessels. In addition to the ganglion and the inner nuclear layers, VEGF protein(s) were located in the endothelial cells of the developing vessels and the angioblasts, i.e., endothelial precursors. On postnatal day 14, when vasculogenesis became inactive, VEGF mRNA expression markedly decreased. These results indicated that VEGF expression in the developing retinas is temporally and spatially correlated with retinal vasculogenesis.

Animals↗
PubMed1994

Astrocytes modulate retinal vasculogenesis: effects on fibronectin expression.

Vasculogenesis is the formation of blood-vessels by differentiation of vascular precursor cells. Experiments using retinal models were designed to test the hypothesis that astrocytes influence this process by effects on the composition of the extracellular matrix. Retinal vasculogenesis was studied in relation to the migration of astrocytes and expression of the extracellular matrix proteins laminin and fibronectin by in vivo experiments in neonatal rats. The results show that astrocytes spread into the retina just ahead of the newly formed vessels, where they probably initiate vasculogenesis. They also establish that fibronectin, but not laminin, is expressed in the zone of vasculogenesis immediately prior to vessel formation. Increased amounts of fibronectin mRNA indicate that fibronectin is synthesized by cells within this same region during this same time period. Later, as the new vessels form, differentiation of endothelial cells is correlated with the appearance of pericytes in the vessel wall and laminin in the vascular basement membrane. In vitro experiments using conditioned medium approaches showed that astrocytes stimulate endothelial cell fibronectin expression. Taken together with the in vivo observations these in vitro results suggest that fibronectin expression is an essential component in the initiation of retinal vasculogenesis. This study is the first indication that astrocytes influence the fibronectin component of the extracellular matrix during retinal vasculogenesis and that expression of fibronectin precedes that of laminin in this process.

Animals↗
PubMed2000

5' nucleotidase and adenosine during retinal vasculogenesis and oxygen-induced retinopathy.

PURPOSE: 5' nucleotidase (5'N) is a major source of the vasogenic substance adenosine in most tissues. The distribution and relative levels of 5'N and adenosine were examined in neonatal dog inner retina during normal vasculogenesis and oxygen-induced retinopathy (OIR). METHODS: Animals ranging in age from 1 to 22 days of age were used in this study. Adenosine immunolocalization was performed on frozen sections with an antibody against adenosine conjugated to laevulinic acid using a streptavidin peroxidase technique. Triplicate room air control animals at different postnatal ages and triplicate oxygen-treated animals at different time points during or after hyperoxic insult were analyzed. Adenosine immunoreactivity (AI) and 5'N enzyme histochemical reaction product were quantified using microdensitometry. Adjacent sections were incubated for von Willebrand factor to label blood vessels. RESULTS: During normal vasculogenesis, AI was most prominent within the inner retina. The peak of immunoreactivity was located at the border of vascularized retina throughout the period of primary retinal vasculogenesis (1-15 days of age). At 22 days when vasculogenesis was complete, AI decreased within the inner retina. The highest 5'N activity was localized to inner Muller cell processes in inner retina and decreased after vasculogenesis was complete. In animals killed after 4 days of oxygen breathing, the vasoobliterative stage of OIR, AI and 5'N activity were reduced throughout the retina. During the vasoproliferative stage, AI was markedly elevated at the edge of reforming vasculature as well as throughout the more posterior inner retina where 5'N activity also was elevated. AI was also in intravitreal neovascularization. CONCLUSIONS: Peak adenosine levels in the inner retina correlated temporally with active vasculogenesis. Adenosine and 5'N levels were reduced in hyperoxia and then returned to above normal levels during the vasoproliferative stage of OIR.

5'-Nucleotidase↗
PubMed2003

Embryonic and adult vasculogenesis.

Two mechanisms account for the formation of blood vessels, vasculogenesis and angiogenesis. Unfortunately, the terms vasculogenesis and angiogenesis literally have the same meaning, i.e., the genesis of blood vessels, and thus do little to distinguish between the two processes. Despite the nomenclature, the two processes are clearly distinct. Vasculogenesis, the de novo formation of blood vessels from mesoderm, is driven by the recruitment of undifferentiated mesodermal cells to the endothelial lineage and the de novo assembly of such cells into blood vessels. Angiogenesis is the generation of new blood vessels from endothelial cells of existing blood vessels, a process driven by endothelial cell proliferation. Recent years have seen dramatic changes in our understanding of the process of vasculogenesis, expanding the scope of its occurrence beyond the earliest stages of development to include involvement in neovascular processes throughout development as well as in the adult. In this review, emphasis is placed on discussion of emerging perspectives on the process of vasculogenesis in both the embryo and the adult.

Adult↗
PubMed2005

Vasculogenesis drives pulmonary vascular growth in the developing chick embryo.

Formation of the pulmonary vasculature has been described as occurring by outgrowth of existing vessels (angiogenesis), de novo formation of new vessels (vasculogenesis), or a combination of both processes. Uncertainty about the contribution of angiogenesis and vasculogenesis to pulmonary vascular formation is partly due to methodologic approaches. Evidence in favor of angiogenesis stems from studies that used vascular-filling methods. Such methods identify only directly continuous lumina. Evidence for vasculogenesis has been provided by the use of molecular markers of blood vessel endothelium. Use of both methods has not been combined in the same species, however. We hypothesized, based on published evidence from quail and mouse, that chick pulmonary vascular formation occurs by vasculogenesis. To test that hypothesis, we used vascular filling, serial section, and immunohistochemical methods to analyze the developing lungs of chick embryos from Hamburger and Hamilton stages 20 to 43. Vascular filling suggested that the lumen of the pulmonary arteries sprouted from the sixth pharyngeal arch arteries. However, serial sections and immunohistochemical localization of fetal liver kinase-1 protein, the receptor for vascular endothelial growth factor, showed that the pulmonary arterial tree formed from endothelial cell precursors and coalescence of isolated blood vessels in the mediastinal splanchnic mesenchyme centrally to the developing lung tissue distally. Pulmonary veins grew from the left atrium to the developing lungs. Pulmonary blood vessel formation occurred continuously throughout the embryonic period studied. Our results show that vasculogenesis is the main process by which the pulmonary vasculature forms in the developing chick embryo.

Animals↗
PubMed2005

Cell elongation is key to in silico replication of in vitro vasculogenesis and subsequent remodeling.

Vasculogenesis, the de novo growth of the primary vascular network from initially dispersed endothelial cells, is the first step in the development of the circulatory system in vertebrates. In the first stages of vasculogenesis, endothelial cells elongate and form a network-like structure, called the primary capillary plexus, which subsequently remodels, with the size of the vacancies between ribbons of endothelial cells coarsening over time. To isolate such intrinsic morphogenetic ability of endothelial cells from its regulation by long-range guidance cues and additional cell types, we use an in vitro model of human umbilical vein endothelial cells (HUVEC) in Matrigel. This quasi-two-dimensional endothelial cell culture model would most closely correspond to vasculogenesis in flat areas of the embryo like the yolk sac. Several studies have used continuum mathematical models to explore in vitro vasculogenesis: such models describe cell ensembles but ignore the endothelial cells' shapes and active surface fluctuations. While these models initially reproduce vascular-like morphologies, they eventually stabilize into a disconnected pattern of vascular "islands." Also, they fail to reproduce temporally correct network coarsening. Using a cell-centered computational model, we show that the endothelial cells' elongated shape is key to correct spatiotemporal in silico replication of stable vascular network growth. We validate our simulation results against HUVEC cultures using time-resolved image analysis and find that our simulations quantitatively reproduce in vitro vasculogenesis and subsequent in vitro remodeling.

Blood Vessels↗
PubMed2002

Vasculogenesis and the search for the hemangioblast.

Embryonic endothelial cells (EC) are generated by two mechanisms, vasculogenesis and angiogenesis (1). The term vasculogenesis describes the de novo emergence of EC progenitors from the mesoderm, whereas angiogenesis corresponds to the generation of EC by sprouting from the pre-existing vascular network. Until recently, it was thought that vasculogenesis was restricted to the period of embryonic development, whereas in the adult, only angiogenesis contributed to EC proliferation. The discovery of circulating EC progenitors in adult bone marrow and peripheral blood has suggested that additional mechanisms besides angiogenesis can occur in the adult, and therefore have renewed interest in the embryonic origin and the development of these progenitor cells. Vasculogenesis in the chick embryo has been studied since the beginning of the 20th century. During early development, vasculogenesis is intimately linked to the emergence of hematopoietic cells (HC). The existence of a common precursor for both EC and HC, termed "hemangioblast," was postulated (2). The purpose of this review is to summarize the experimental evidence concerning the emergence of EC and HC during embryonic life.

Adult↗
PubMed2004

Apoptosis contributes to vascular lumen formation and vascular branching in human placental vasculogenesis.

Placental vasculogenesis consists of several stages, including appearance of hemangioblasts and angiogenic cell islands, setting up a primitive vascular network, and transition from vasculogenesis to sprouting and nonsprouting angiogenesis. In the present study, we hypothesized that placental vasculogenesis and angiogenesis require apoptosis during the formation of primitive vascular pattern, vessel elongation, and angiogenic branching. Vasculogenesis and apoptotic cells were identified using CD31 immunohistochemistry, hematoxylin-eosin (H-E) staining, CD31-TUNEL double-labeling, and transmission-electron microscopy (TEM). No TUNEL-positive cell was detected in angiogenic cell islands; however, several TUNEL-positive cells were observed during the primitive lumen formation. Interestingly, some of the stromal cells located between vasculogenic areas during the endothelial tube elongation and angiogenic branching also were TUNEL-positive. The presence of morphological aspects of apoptosis, such as nuclear shrinkage and nuclear bodies (apoptotic bodies), also was confirmed in H-E-stained and TEM-depicted sections. Quantitative analysis showed that higher ratios for apoptotic cells were found in the core stroma of villi among the vascular branching areas and in the primitive capillary lumen compared to angiogenic cell cords and vasculatures with advanced lumens (P < 0.05). In conclusion, our results suggest that apoptosis likely is involved in the physiologic mechanisms of placental vasculogenesis and angiogenesis, such as lumen formation and angiogenic branching.

Adult↗
PubMed2006

VEGF and PlGF promote adult vasculogenesis by enhancing EPC recruitment and vessel formation at the site of tumor neovascularization.

There are growing data to suggest that tissue hypoxia represents a critical force that drives adult vasculogenesis. Vascular endothelial growth factor (VEGF) expression is dramatically up-regulated by hypoxia and results in enhanced neovascularization. Although the role of VEGF in angiogenesis has been well characterized, its role in adult vasculogenesis remains poorly understood. We used two distinct murine bone marrow transplantation (BMT) models to demonstrate that increased VEGF levels at the site of tumor growth promoted vasculogenesis in vivo. This effect of VEGF was downstream of its effect to enhance either mobilization or survival of circulating endothelial progenitor cells (EPCs). Both VEGFR1 (flt1) and VEGFR2 (flk1) are expressed on culture expanded human EPCs. Previous studies suggest that the effect of VEGF on endothelial cell migration is primarily mediated via VEGFR2; however, VEGF-induced EPC migration in vitro was mediated by both receptors, suggesting that VEGF-VEGFR1 interactions in EPCs are distinct from differentiated endothelial cells. We used specific blocking antibodies to these receptors to demonstrate that VEGFR1 plays an important role in human EPC recruitment to tumors. These findings were further supported by our finding that tumor-associated placental growth factor (PlGF), a VEGFR1-specific agonist, increased tumor vasculogenesis in a murine BMT model. We further showed that both VEGF receptors were necessary for the formation of functional vessels derived from exogenously administered human ex vivo expanded EPCs. Our data suggest local VEGF and/or PlGF expression promote vasculogenesis; VEGF plays a role in EPC recruitment and subsequent formation of functional vessels.

Animals↗
PubMed2006

Vasculogenesis of the embryonic heart: contribution of nucleated red blood cells to early vascular structures.

During embryogenesis, coronary vessels develop via vasculogenesis and angiogenesis. Vasculogenesis is formation in situ of primary vessels from angioblasts - endothelial cell progenitors, and angiogenesis is formation of vessels from the existing ones. In the embryonic heart vasculogenesis precedes and overlaps angiogenesis and lasts till the end of the fetal life. What is unique about heart vasculogenesis is the fact that nucleated blood cells accompany early angioblasts in a spatiotemporal way. Morphologically these structures resemble yolk sac blood islands, thus, they have been called blood-island-like structures. In addition, these early vascular structures (blood-island-like) are found in the heart before coronary vessel system connects with the systemic circulation. We present the recent data regarding endothelial cell properties and derivation during coronary vessel formation and hypotheses concerning a source of blood cells in early vascular structures of the heart; the latter has received little attention in the literature. This review summarizes current knowledge on the endothelial cell origination from epicardial mesothelium or liver primordium. This review also focuses on blood cell contribution to coronary vessel vasculogenesis. The role of proepicardium in the epicardial cover formation and the epicardium as a source of cellular components of coronary vasculature and interstitial fibroblasts is presented. It seems that blood cells and angioblasts, which form the early vascular structures do not derive from the same hemangioblastic precursor.

Animals↗
PubMed1992

Antibodies to beta 1-integrins cause alterations of aortic vasculogenesis, in vivo.

Vasculogenesis is the de novo formation of blood vessels from mesoderm. This process occurs very early in development and provides a convenient system for studying morphogenesis in higher vertebrates. The cell-extracellular matrix (ECM) interactions that occur during dorsal aortic vasculogenesis were examined using the monoclonal antibody, CSAT, a reagent known to neutralize the ligand-binding activity of avian beta 1-integrins. We injected CSAT into quail embryos during a period of active vasculogenesis (4-10 somites). The CSAT antibodies, but not controls, had a marked and reproducible effect on aortic vessel formation. Vasculogenesis appeared to be arrested at the stage when slender cord-like assemblies of angioblasts rearrange to form tubules. Indeed, aortic primordia near the site of CSAT injection did not form patent vessels.

Animals↗
PubMed2005

Genes critical to vasculogenesis as defined by systematic analysis of vascular defects in knockout mice.

To identify genes important to the process of vasculogenesis, we evaluated embryonic vascular anomalies from 100 mouse knockout studies using a novel meta-analysis approach. By applying this method, termed approach for ranking of embryonic vascular anomalies (AREVA), rank scores were calculated for each knockout based on the occurrence of vascular defects during periods of vasculogenesis in specific embryonic regions. As a result, 12 genes (fibronectin, VEGFR-1/Flt-1, VEGFR-2/Flk-1, alpha 5 integrin, Tek/Tie2, VE-cadherin, VEGFA, connexin 45, ShcA, cytochrome P450 reductase, CD148/DEP-1, and EphrinB2) were determined to play critical roles in vasculogenesis. Functional categorization of these genes revealed the fundamental importance of VEGF signaling since 10 of the 12 genes (fibronectin, VEGFR-1/Flt-1, VEGFR-2/Flk-1, alpha 5 integrin, VE-cadherin, VEGFA, ShcA, cytochrome P450 reductase, CD148/DEP-1, and EphrinB2) relate to this pathway. Furthermore, the findings highlight a potential network for regulating VEGF signaling involving integration of fibronectin, EphrinB2, Tie2, and connexin 45 signaling pathways via the ShcA/Ras/Raf/Mek/Erk cascade. In addition to retrospective application of AREVA as done herein, AREVA can be used prospectively to determine the relevancy to vasculogenesis of newly inactivated genes.

Adaptor Proteins, Signal Transducing↗
PubMed1994

Induction of vasculogenesis in quail blastodisc-derived embryoid bodies.

We have previously developed an in vitro culture system in which dissociated cells from unincubated quail blastodiscs formed in vivo-like blood islands consisting of blood and endothelial cells in response to fibroblast growth factors (FGFs). Here we demonstrate that the same quail blastodisc cells grown in suspension culture in the presence of basic FGF (bFGF) reaggregated and formed three-dimensional spherules (embryoid bodies, EBs) which underwent vasculogenesis and hematopoiesis within 3 days. In contrast to murine embryoid bodies, which undergo vasculogenesis spontaneously, the formation of vascular structures in quail blastodisc cultures was absolutely dependent on bFGF. While about 75% contained blood islands and about 50% formed capillaries in the presence of bFGF, only 0.2% of the embryoid bodies formed blood islands in control cultures without bFGF. Vascular channels were gradually encoated by primitive smooth muscle cells within 5 days. Ultrastructural examinations revealed capillary blood vessels and blood islands indistinguishable from their yolk sac counterparts. Mesodermal tissue was present in cultures both with and without bFGF, but consisted of an avascular undifferentiated mesenchyme in control cultures. Since the entire sequence of vasculogenesis from the formation of endothelial cells to their assembly into a vascular plexus is observed in response to the inducer bFGF, this culture system is a suitable model for studying the molecular events that initiate the emergence of endothelial cells and the formation of a vascular plexus during vasculogenesis.

Animals↗
PubMed1995

Vascular endothelial growth factor (VEGF) and VEGF receptor 2 (flk-1) are expressed during vasculogenesis and vascular differentiation in the quail embryo.

Vasculogenesis, the de novo formation of embryonic blood vessels from their angioblastic precursors in situ, is supposed to be under the control of polypeptide growth factors and their receptors. The receptor tyrosine kinase flk-1 and its high-affinity ligand vascular endothelial growth factor (VEGF) represent an endothelial specific signal transduction system expressed during embryonic vascular growth in the mouse. We have cloned the quail homologs of VEGF and flk-1 using PCR and have investigated their expression pattern in vivo. As shown by Northern analysis and reverse transcription PCR, VEGF and flk-1 mRNA (3.9 and 5.8 kb, respectively) were already present in the unincubated blastodisc at low levels and were largely upregulated during gastrulation at Embryonic Day 1. As detected by in situ hybridization, flk-1 mRNA was initially present in the entire mesoderm of Day 1 embryos but from Day 2 on was restricted to endothelial cells. At Day 2 VEGF was ubiquitously expressed in the embryo proper and was mainly restricted to the vascularized part (area vasculosa) in the yolk sac. Later on VEGF expression was detected in all organs. In the kidney VEGF mRNA was mainly localized to the glomeruli. This pattern of expression is consistent with the pattern found during mouse embryogenesis. We have recently established an in vitro model of vasculogenesis in which hemangioblastic precursors are induced in cell cultures from the unincubated quail blastodisc by basic fibroblast growth factor (bFGF) and give rise to blood vessels in vitro. Taking advantage of this in vitro model we examined whether FGF and VEGF act in concert during vasculogenesis. We found that the flk-1 receptor mRNA is dramatically upregulated within 24 hr upon the addition of FGF to quail blastodisc cell cultures. This inducibility in response to FGF is confined to the first 24 hr of culture. The early expression of the flk-1 mRNA may characterize the differentiation of hemangioblastic precursors from pluripotent epiblast cells which in vivo is initiated during gastrulation. Thus, the time course and the pattern of expression during embryogenesis in different species suggest a major role for the VEGF/flk-1 signal transduction system in vasculogenesis and angiogenesis.

Amino Acid Sequence↗
PubMed1996

Induction of embryonic vasculogenesis by bFGF and LIF in vitro and in vivo.

The de novo formation of blood vessels (vasculogenesis) is an integral part of embryogenesis. Elucidation of the role of cytokine cooperation in vasculogenesis may lead to a better understanding of organogenesis, blood vessel regulation during tumorigenesis, and tissue injury. We have used embryonic stem cells to derive an endothelial cell line, designated IEM, which expresses a range of endothelial markers, including Von Willibrand Factor VIII related antigen, vascular cell adhesion molecule, platelet-endothelial cell adhesion molecule (CD31), and receptors for acetylated low-density lipoprotein. More importantly, IEM cells can be induced upon exposure to combinations of basic fibroblast growth factor and leukemia inhibitory factor (LIF) to proliferate and undergo vasculogenesis in vitro, resulting in the formation of vascular tubes and microcapillary anastomoses. Moreover, exposure to both cytokines conditionally permits IEM cells to specifically chimerize microvascular endothelium in vivo following blastocyst injection. These results indicate that bFGF and LIF together contribute to the induction and support of embryonic vasculogenesis in an isolated endothelial cell line. Our results provide evidence that combined actions of bFGF/LIF may play a role in mechanisms controlling blood vessel development.

Animals↗
PubMed2001

In vitro model for developmental progression from vasculogenesis to angiogenesis with a murine endothelial precursor cell line, MFLM-4.

In the embryo, vascular networks are developed through both vasculogenesis, the assembly of vessels from endothelial progenitor cells or hemangioblasts, and angiogenesis, the sprouting of vessels from preexisting capillaries. Cell culture models using endothelial cells (EC) and various extracellular matrix components have been useful in understanding the cellular and molecular factors involved in angiogenesis. However, there are few models of vasculogenesis. Using a murine endothelial precursor cell line, MFLM-4, derived from e14.5 lung mesenchyme, we have developed a culture system that not only recapitulates many of the characteristics of vasculogenesis but also progresses into angiogenesis. By 8 h, MFLM-4 cultured on the basement membrane preparation Matrigel invade the matrix, coalesce, and assemble into large clusters of cells resembling blood islands. During vascular development, blood islands are the focal areas for coalescence of endothelial precursors. For MFLM-4, this phase of in vitro vasculogenic clustering does not require proliferation. If proliferation is not blocked, MFLM-4 progresses into an angiogenic phase with the clusters forming multicell angiogenic sprouts. Through 3 days of culture, lumens form within the clusters, adjacent clusters are connected with tube-like structures, and eventually an extensive network or plexus of clusters connected by capillary-like tubes is formed. MFLM-4 cultured on Matrigel provides an in vitro system for analysis of the multistage, concurrent processes of vasculogenesis and angiogenesis.

Animals↗
PubMed2005

Sequential steps during vasculogenesis and angiogenesis in the very early human placenta.

Development of blood vessels takes place via two subsequent processes, vasculogenesis and angiogenesis. During vasculogenesis, formation of first blood vessels is achieved by differentiation of hemangiogenic stem cells from pluripotent mesenchymal cells, while during angiogenesis new blood vessels form from already existing vessels. The combination of our data with those from the literature leads us to depict the chronological steps of cell differentiation in the mesenchymal core of placental villi during vasculogenesis and angiogenesis. This current opinion will focus on the temporal and spatial expression of VEGF and its receptors VEGFR-1 and VEGFR-2, and the angiopoietin receptors Tie-1 and Tie-2 in parallel to vascular maturation in human placental villi during very early stages of placental development. There is evidence that the interplay of a variety of growth factors secreted from different cell types during development is needed to trigger as well as maintain placental vasculogenesis and angiogenesis.

Adult↗
PubMedDate not supplied

Defective vasculogenesis in systemic sclerosis.

BACKGROUND: Typical vascular features of systemic sclerosis include low capillary density and vascular obliteration. The formation and repair of blood vessels in adults involve vasculogenesis, which is mediated through the recruitment of bone-marrow-derived circulating endothelial precursors (CEP). We investigated whether vasculogenesis is impaired in patients with systemic sclerosis. METHODS: Peripheral blood was obtained from 11 patients with systemic sclerosis, 11 with rheumatoid arthritis, and 11 healthy controls. Factors potentially affecting the CEP number were matched among the three groups. CEP (identified as circulating cells positive for CD34, CD133, and the type 2 receptor for vascular endothelial growth factor) were quantified by cell sorting and three-colour flow cytometry. The circulating concentrations of angiogenic factors were measured by ELISA. The potential of CEP to differentiate into endothelial cells was assessed by the upregulation of von Willebrand factor after in-vitro maturation treatment. Findings The absolute number of CEP was much lower in patients with systemic sclerosis than in patients with rheumatoid arthritis or healthy controls (median 274 [IQR 178-395] vs 1154 [653-1524] and 1074 [713-1186] per 20 mL peripheral blood, respectively; p<0.0001 by Kruskal-Wallis test. Paradoxically, circulating concentrations of most angiogenic factors were significantly higher in patients with systemic sclerosis than in healthy controls. The proportion of CEP that differentiated into endothelial cells was significantly lower in patients with systemic sclerosis than in healthy controls (p<0.0001, Mann-Whitney test). INTERPRETATION: Insufficient vascular repair machinery due to defective vasculogenesis might contribute to vasculopathy in systemic sclerosis. RELEVANCE TO PRACTICE: As well as providing an important insight into the pathogenesis of this disorder, these findings suggest that dysregulated vasculogenesis might be important in other disorders with abnormalities in vascular formation, including those with excessive formation of new vessels such as cancer and those with deficient vessel formation such as atherosclerosis. Circulating endothelial precursors could be a novel target for therapeutic strategies for ischaemic complications in patients with systemic sclerosis.

AC133 Antigen↗