AMD3100 Mobilizes Endothelial Progenitor Cells in Mice, But Inhibits Its Biological Functions by Blocking an Autocrine/Paracrine Regulatory Loop of Stromal Cell Derived Factor-1 In Vitro

Yin, Yi; Huang, Lan; Zhao, Xinshu; Fang, Yuqiang; Yu, Shiyong; Zhao, Jinghong; Cui, Bing

doi:10.1097/fjc.0b013e3180587e4d

“…Although long-term treatments with Cxcr4 antagonists have been shown to stimulate EPC mobilization in mice, 53 these antagonists also interfered with the adhesion, migration, and proliferation capacity of EPCs in vitro. 54 This can explain why systemic treatment of mice with Cxcr4 antagonists did not affect the reendothelialization degree after carotid artery injury in mice, 9,55,56 in contrast to BmxCreER T2 -mediated EC-specific Cxcr4 deficiency studied in this article.…”

Section: Cd31mentioning

confidence: 85%

Deficiency of Endothelial Cxcr4 Reduces Reendothelialization and Enhances Neointimal Hyperplasia After Vascular Injury in Atherosclerosis-Prone Mice

Noels

¹

,

Zhou

²

,

Tilstam

³

et al. 2014

ATVB

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Objective— The Cxcl12/Cxcr4 chemokine ligand/receptor axis mediates the mobilization of smooth muscle cell progenitors, driving injury-induced neointimal hyperplasia. This study aimed to investigate the role of endothelial Cxcr4 in neointima formation. Approach and Results— β-Galactosidase staining using bone marrow x kinase ( Bmx ) -CreER T2 reporter mice and double immunofluorescence revealed an efficient and endothelial-specific deletion of Cxcr4 in Bmx-CreER T2+ compared with Bmx-CreER T2− Cxcr4-floxed apolipoprotein E–deficient ( Apoe −/− ) mice (referred to as Cxcr4 EC-KO ApoE −/− and Cxcr4 EC-WT ApoE −/− , respectively). Endothelial Cxcr4 deficiency significantly increased wire injury–induced neointima formation in carotid arteries from Cxcr4 EC-KO ApoE −/− mice. The lesions displayed a higher number of macrophages, whereas the smooth muscle cell and collagen content were reduced. This was associated with a significant reduction in reendothelialization and endothelial cell proliferation in injured Cxcr4 EC-KO ApoE −/− carotids compared with Cxcr4 EC-WT ApoE −/− controls. Furthermore, stimulation of human aortic endothelial cells with chemokine (C-X-C motif) ligand 12 (CXCL12) significantly enhanced their wound-healing capacity in an in vitro scratch assay, an effect that could be reversed with the CXCR4 antagonist AMD3100. Also, flow cytometric analysis showed a reduced mobilization of Sca1 + Flk1 + Cd31 + and of Lin − Sca1 + progenitors in Cxcr4 EC-KO ApoE −/− mice after vascular injury, although Cxcr4 surface expression was unaltered. No differences could be detected in plasma concentrations of Cxcl12, vascular endothelial growth factor, sphingosine 1-phosphate, or Flt3 (fms-related tyrosine kinase 3) ligand, all cytokines with an established role in progenitor cell mobilization. Nonetheless, double immunofluorescence revealed a significant reduction in local endothelial Cxcl12 staining in injured carotids from Cxcr4 EC-KO ApoE −/− mice. Conclusions— Endothelial Cxcr4 is crucial for efficient reendothelialization after vascular injury through endothelial wound healing and proliferation, and through the mobilization of Sca1 + Flk1 + Cd31 + cells, often referred to as circulating endothelial progenitor cells.

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“…Doses of 5 to 10 mg/kg are routinely administered to adult mice for acute mobilization of hematopoietic progenitor cells. 28,43,44 BecauseAMD3100 has a short half-life in vivo (3.6 hours in healthy humans 45 ), an increased dose was used to antagonize CXCR4 activity long enough to effect a change in the developing retinal vasculature. Although AMD3100 has been reported to be highly specific to CXCR4, a recent report suggests that it may also bind the other known SDF-1 receptor, CXCR7.…”

Section: Discussionmentioning

confidence: 99%

“…[28][29][30] SDF-1 also promotes endothelial tube formation. [31][32][33] We examined the effects of SDF-1 on capillary formation and outgrowth in vitro.…”

Section: Sdf-1 Promotes Capillary Formation In Vitromentioning

confidence: 99%

Microarray analysis of retinal endothelial tip cells identifies CXCR4 as a mediator of tip cell morphology and branching

Strasser¹,

Kaminker²,

Tessier‐Lavigne³

2010

Blood

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The development of the vertebrate vascular system is mediated by both genetic patterning of vessels and by angiogenic sprouting in response to hypoxia. Both of these processes depend on the detection of environmental guidance cues by endothelial cells. A specialized subtype of endothelial cell known as the tip cell is thought to be involved in the detection and response to these cues, but the molecular signaling pathways used by tip cells to mediate tissue vascularization remain largely uncharacterized. To identify genes critical to tip cell function, we have developed a method to isolate them using laser capture microdissection, permitting comparison of RNA extracted from endothelial tip cells with that of endothelial stalk cells using microarray analysis. Genes enriched in tip cells include ESM-1, angiopoietin-2, and SLP-76. CXCR4, a receptor for the chemokine stromal-cell derived factor-1, was also identified as a tip cell-enriched gene, and we provide evidence for a novel role for this receptor in mediating tip cell morphology and vascular patterning in the neonatal retina. IntroductionDuring embryogenesis, the vertebrate circulatory system develops through the overlapping processes of vasculogenesis and angiogenesis. Vasculogenesis is the migration and differentiation of endothelial cells from hematopoietic precursors. Angiogenesis involves the sprouting of blood vessel networks and their maturation into an organized, functional circulatory system. 1,2 Many aspects of angiogenesis are also recapitulated in the adult during normal conditions, such as pregnancy, and pathologic situations, such as inflammation and cancer. 3 An understanding of the molecular mechanisms that govern this process is critical for the development of novel proangiogenic and antiangiogenic therapeutics.A specialized subset of endothelial cells known as tip cells is thought to mediate vessel growth and patterning. Tip cells are found at the growing end of a nascent vascular sprout and are characterized by abundant and dynamic filopodia. These filopodia are thought to detect cues in the local environment and translate them into directed motility. 4,5 Tip cells are responsive to angiogenic factors, such as vascular endothelial growth factor-A (VEGF-A), 6 as well as classic axon guidance factors, such as netrin-1. 7 The generation of new tip cells is limited by the activity of Delta-like 4 (Dll4), [8][9][10][11] which is specifically up-regulated in tip cells. 12 Signaling of Dll4 through Notch receptors on adjacent cells has been proposed to inhibit their development into tip cells. 13 Despite these advances, the mechanisms that may promote or mediate the generation of new tip cells remain incompletely understood.Recently, endothelial tip cells have become a topic of intense scrutiny, 3 but many critical questions remain about their function and how they may differ from neighboring stalk cells. Expression of several genes, including VEGF receptor 2 (VEGFR2), 6 Dll4, PDGF-B, 6 and VEGFR3, 14 has been shown to be higher in retinal tip cel...

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“…To this end, SDF-1a binding to its specific receptors CXCR4 and CXCR7 32,33 was blocked by addition of AMD3100 in different concentrations (0-10 mg/mL). [34][35][36][37] Although the total length and size of the prevascular structures was not significantly altered when AMD3100 was present in (A, E). MSC/endothelial progenitor cell co-culture pellets were maintained in either 1:1 osteogenic/ endothelial medium (B, F); endothelial medium (C, G); or osteogenic medium (D, H).…”

Section: Involvement Of Sdf-1 Signaling In In Vitro Vasculogenesismentioning

confidence: 99%

Hypoxia Impedes Vasculogenesis of In Vitro Engineered Bone

Gawlitta

¹

,

Fledderus

²

,

Rijen

³

et al. 2012

Tissue Engineering Part A

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To ensure the survival of engineered bone after implantation, we combined human endothelial colony forming cells (ECFCs) and multipotent stromal cells (MSCs) as a proof of concept in a co-culture model to create in vitro prevascularized bone constructs. We hypothesized that a hypoxic stimulus will contribute to prevascularization of engineered bone. Bone marrow-derived MSCs and ECFCs from human adult peripheral blood were allowed to form co-culture pellets containing ECFCs and MSCs (1:4) or MSCs only in controls. After culture under normoxia or hypoxia (5%), pellets were harvested and processed for immunohistochemistry of CD31, a-smooth muscle actin, and osteocalcin. Expression of vascular endothelial growth factor and SDF-1a was analyzed by PCR to elucidate their involvement in hypoxic stimulation of prevascularization. The normoxic condition in cocultures of MSCs and ECFCs supported the formation and maintenance of prevascular structures, including organized CD31-positive cells embraced by differentiated mural cells. These structures failed to form in hypoxic conditions, thereby rejecting the hypothesis that hypoxia stimulates prevasculogenesis in three-dimensional engineered bone constructs. Further, the formation of prevascular structures was paralleled by increased SDF-1a expression. It is suggested that actual oxygen levels were below 5% in the hypoxic co-cultures, which prevented prevascular structure formation. In conclusion, our normoxic co-culture model containing cells from clinically relevant sources sustained simultaneous endothelial, smooth muscle, and osteogenic differentiation.

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AMD3100 Mobilizes Endothelial Progenitor Cells in Mice, But Inhibits Its Biological Functions by Blocking an Autocrine/Paracrine Regulatory Loop of Stromal Cell Derived Factor-1 In Vitro

Cited by 47 publications

References 31 publications

Deficiency of Endothelial Cxcr4 Reduces Reendothelialization and Enhances Neointimal Hyperplasia After Vascular Injury in Atherosclerosis-Prone Mice

Deficiency of Endothelial Cxcr4 Reduces Reendothelialization and Enhances Neointimal Hyperplasia After Vascular Injury in Atherosclerosis-Prone Mice

Microarray analysis of retinal endothelial tip cells identifies CXCR4 as a mediator of tip cell morphology and branching

Hypoxia Impedes Vasculogenesis of In Vitro Engineered Bone

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