Mitogenic and<i>In Vitro</i>Angiogenic Activity of Human Recombinant Heparin Affin Regulatory Peptide

Laaroubi, K.; Delbé, Jean; Vacherot, Francis; Desgranges, Pascal; Tardieu, M; Jaye, Michael; Barritault, Denis; Courty, José

doi:10.3109/08977199409010982

Cited by 92 publications

(90 citation statements)

References 30 publications

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“…Sequence analysis of this HARP isolated by Mono S chromatography showed that it had the N-terminal sequence NH 2 -Gly-Lys-Lys-Glu-Lys-Pro. Comparison with the N-terminal sequence of the mitogenic form supports the data that the mitogenicity of HARP is directly or indirectly related to the presence of three N-terminal amino acids (22).…”

Section: Discussionsupporting

confidence: 58%

“…The cells were grown in DMEM supplemented with 10% fetal calf serum in the presence of 5 ng/ml FGF2 and were used between passages 10 and 20. NIH3T3 cell lines stably transfected with pJK12 that overexpressed HARP (H-NIH3T3) were cultured as described previously (22). MDA-MB 231 cells were a gift from M. Crépin (Bobigny, France), and MC 3T3-E1 cells were a gift from M. Meunier (Lyon, France).…”

Section: Materials-cellmentioning

confidence: 99%

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Glycosaminoglycans Differentially Bind HARP and Modulate Its Biological Activity

Vacherot

Delbé

Héroult

et al. 1999

Journal of Biological Chemistry

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Heparin affin regulatory peptide (HARP) is a polypep-) and a lower affinity (K d ‫؍‬ 51 nM). Exogenous heparin, heparan sulfate, and dermatan sulfate potentiated the growth-stimulatory activity of HARP, suggesting that corresponding proteoglycans could be involved in the regulation of the mitogenic activity of HARP. Heparin affin regulatory peptide (HARP)1 belongs to a growing group of heparin binding extracellular regulatory molecules, and it has mitogenic and neurite outgrowth activities.Independently purified from perinatal rat brain as a polypeptide that induces neurite outgrowth of embryonic neurons (1), this protein, also named pleiotrophin (2), has been purified from uterus (3) and adult brain (4) as a growth factor for fibroblastic and endothelial cells. Expression of HARP mRNA is developmentally regulated, and major expression is found during the perinatal growth period in rat brain (5, 6), pointing to the possible function of this molecule in the maturation of nerve cells. Moreover, HARP mRNA is found in several adult tissues, and HARP has been implicated in tumor growth (7).HARP cDNA has been cloned and sequenced from several species. The predicted amino acid sequence has been determined in humans, mice, and rats (2, 6, 8, 9) and shows 98% homology among the three species. In addition, the HARP amino acid sequence shares 55% homology with the midkine gene product (10). In contrast to their neurite outgrowth activity, initial studies differed in their results about the mitogenic activity of midkine and HARP (3, 9, 11-15), possibly because of differences in the cell type used or to the isolation procedure.More recent studies clearly demonstrate that this family of polypeptides have growth-stimulatory activity (16).The high affinity of HARP for heparin suggests that HARP may bind to heparan sulfate proteoglycans (HSPGs) present in extracellular compartments defined as cell surface and extracellular matrix (ECM). Recent studies have demonstrated that syndecan-1, syndecan-3 (N-syndecan), and syndecan-4 (ryudocan) bind HARP with high affinity (13,17,18). In addition, biochemical and cell biological studies have pointed to syndecan-3 as the HARP receptor involved in neurite outgrowth activity (19). Despite the correlation derived from previous studies, there has been no direct biochemical demonstration that HARP is trapped in the extracellular compartment as a mitogenic molecule bound to HSPGs. In this study we investigated whether HARP is present in the extracellular compartment as well as the role of glycosaminoglycans (GAGs) in controlling HARP mitogenic activity. (Proteus vulgaris; EC 4.2.2.4), leupeptin, pepstatin, phenylmethylsulfonyl fluoride, heparan sulfate (HS) from bovine intestinal mucosa, keratan sulfate (KS) from bovine cornea, chondroitin sulfate A (CS-A) from bovine trachea, dermatan sulfate (DS) from porcine skin, and chondroitin sulfate C (CS-C) from shark cartilage were purchased from Sigma. Horseradish peroxidase-conjugated goat anti-rabbit immunoglobulins were obtained from Diagnostics-...

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Section: Discussionsupporting

confidence: 58%

Section: Materials-cellmentioning

confidence: 99%

Glycosaminoglycans Differentially Bind HARP and Modulate Its Biological Activity

Vacherot

Delbé

Héroult

et al. 1999

Journal of Biological Chemistry

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“…Compelling evidence that HARP contributes to the regulation of angiogenesis has been reported (Laaroubi et al, 1994;Choudhuri et al, 1997;Delbe et al, 1999). Moreover, both HARP protein and its mRNA have been identified in blood vessels and capillaries in mammary gland (Ledoux et al, 1997), prostate (Vacherot et al, 1999), and uterus (Milhiet et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Heparin affin regulatory peptide binds to vascular endothelial growth factor (VEGF) and inhibits VEGF-induced angiogenesis

et al. 2004

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Heparin affin regulatory peptide (HARP) is an heparinbinding molecule involved in the regulation of cell proliferation and differentiation. Here, we report that HARP inhibited the biological activity induced by the 165-amino-acid form of vascular endothelial growth factor (VEGF 165 ) on human umbilical vein endothelial cells. Endothelial-cell proliferation induced by VEGF 165 showed about 50% inhibition in the presence of HARP in a concentration of 3 nM. In similar range of concentrations, HARP blocked tube formation induced by VEGF 165 in three-dimensional angiogenesis assay. In vivo studies showed that HARP inhibited the VEGF 165 -induced Matrigelt infiltration of endothelial cells. We then investigated the mechanisms of this inhibition and shown that HARP inhibited the binding of 125 I-VEGF 165 to the VEGF receptors of endothelial cells. Additional studies using VEGF soluble receptors indicated that binding of 125 I-VEGF 165 to kinase insert domain-containing receptor and neuropilin receptor was inhibited by HARP, but conversely the binding of 125 I-VEGF 165 to fms-like tyrosine kinase I receptor was unaffected. A competitive affinity-binding assay demonstrated that HARP interacted directly with VEGF 165 with a dissociation coefficient of 1.38 nM. Binding assay using deletion mutants of HARP revealed that the thrombospondin type-1 repeats domains were involved in this interaction. These data demonstrate for the first time that the angiogenic factor HARP can also negatively regulates the angiogenic activity of VEGF 165 .

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“…More recently, new growth factor members of the heparin-binding growth factor family such as pleiotrophin (PTN) and midkine (MDK) have been described, which also contribute to promoting tumorigenesis and angiogenesis. PTN was shown to induce the proliferation and migration of endothelial cells in vitro (2 -6) and has been correlated with enhanced tumor growth and vascular density in vivo (5,6). PTN was shown to be particularly involved in tumor growth of several cancer types (4,7) and to also display transforming activity (8).…”

Section: Introductionmentioning

confidence: 99%

16-kDa fragment of pleiotrophin acts on endothelial and breast tumor cells and inhibits tumor development

Ducès

Karaky

Martel-Renoir

et al. 2008

Molecular Cancer Therapeutics

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Pleiotrophin (PTN) is a 136-amino acid secreted heparinbinding protein that is considered as a rate-limiting growth and an angiogenic factor in the onset, invasion, and metastatic process of many tumors. Its mitogenic and tumorigenic activities are mediated by the COOH-terminal residues 111 to 136 of PTN, allowing it to bind to cell surface tyrosine kinase-linked receptors. We investigated a new strategy consisting in evaluating the antitumor effect of a truncated PTN, lacking the COOH-terminal 111 to 136 portion of the molecule (PTN#111-136), which may act as a dominant-negative effector for its mitogenic, angiogenic, and tumorigenic activities by heterodimerizing with the wild-type protein. In vitro studies showed that PTN#111-136 selectively inhibited a PTN-dependent MDA-MB-231 breast tumor and endothelial cell proliferation and that, in MDA-MB-231 cells expressing PTN#111-136, the vascular endothelial growth factor-A and hypoxia-inducible factor-1A mRNA levels were significantly decreased by 59% and 71%, respectively, compared with levels in wild-type cells. In vivo, intramuscular electrotransfer of a plasmid encoding a secretable form of PTN#111-136 was shown to inhibit MDA-MB-231 tumor growth by 81%. This antitumor effect was associated with the detection of the PTN#111-136 molecule in the muscle and tumor extracts, the suppression of neovascularization within the tumors, and a decline in the Ki-67 proliferative index. Because PTN is rarely found in normal tissue, our data show that targeted PTN may represent an attractive and new therapeutic approach to the fight against cancer.

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Mitogenic andIn VitroAngiogenic Activity of Human Recombinant Heparin Affin Regulatory Peptide

Cited by 92 publications

References 30 publications

Glycosaminoglycans Differentially Bind HARP and Modulate Its Biological Activity

Glycosaminoglycans Differentially Bind HARP and Modulate Its Biological Activity

Heparin affin regulatory peptide binds to vascular endothelial growth factor (VEGF) and inhibits VEGF-induced angiogenesis

16-kDa fragment of pleiotrophin acts on endothelial and breast tumor cells and inhibits tumor development

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