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

Ducès, Aurélie; Karaky, Racha; Martel-Renoir, Dominique; Mir, Lluis M.; Hamma-Kourbali, Yamina; Bièche, Ivan; Opolon, Paule; Delbé, Jean; Courty, José; Perricaudet, Michel; Griscelli, Frank

doi:10.1158/1535-7163.mct-08-0301

Cited by 23 publications

(32 citation statements)

References 39 publications

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“…The pcDNA3-LacZ control plasmid encoding b-galactosidase was constructed as previously described (10). The pcDNA3-MDKD81-121-MycHis plasmid was obtained by cloning the MDKD81-121 fragment between the BamHI and the EcoRI sites of the pcDNA3.1(þ)/myc-HisA plasmid (Invitrogen).…”

Section: Plasmid Dnamentioning

confidence: 99%

“…Human umbilical smooth muscle cells (SMC) were a kind gift of Dr. Georges Uzan (INSERM U972, Hôpital PaulBrousse, Villejuif, France) and were isolated and characterized as described previously (22). Human umbilical vein endothelial cells (HUVEC, Lonza CC-2517) and SMCs were cultured in their adequate medium from Lonza and promocell, respectively (10). Experiments involving ATCC or Lonza cell lines were mostly performed in the same year they were purchased.…”

Section: Cell Culture and Conditioned Mediummentioning

confidence: 99%

“…PTN expression in neuroblastoma cell pellets was assessed as previously described (10). For MDK detection in cell pellets by Western blot analysis, proteins were transferred to a nitrocellulose membrane (Pall) that was blocked in PBS 0.2% Tween and 5% powdered milk.…”

Section: Western Blot Analysis Elisa and Fishmentioning

confidence: 99%

“…Swain anti-rabbit peroxidase-linked antibody (1:1,000; Dako) was used as secondary antibody. ALK Western blot was performed as previously described (10). For the RPTPb/z Western blot analysis, the protein was detected using a primary rat anti-RPTPb/z antibody (1:500, R&D Systems), and a secondary anti-rat antibody.…”

Section: Western Blot Analysis Elisa and Fishmentioning

confidence: 99%

“…They are capable of silencing the native protein by forming inactive heterodimeric molecules (9). In particular, we demonstrated that a truncated Pleiotrophin (PTN), lacking the C-terminal 111-136 portion of the molecule (PTND111-136), acted as a dominant-negative mutant that selectively inhibited in vitro the proliferation of a PTN-dependent MDA-MB-231 breast tumor and endothelial cells by silencing the VEGF-A and HIF1a pathways and breast tumor growth in vivo by 81% (10).…”

Section: Introductionmentioning

confidence: 99%

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Midkine Lacking Its Last 40 Amino Acids Acts on Endothelial and Neuroblastoma Tumor Cells and Inhibits Tumor Development

Dianat

Viet

Gobbo

et al. 2015

Molecular Cancer Therapeutics

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Midkine (MDK) is a member of a new family of neurotrophic factors considered as rate-limiting growth and angiogenic factors implicated in the onset, invasion, and metastatic process of neuronal tumors, including neuroblastoma. We showed that all neuroblastoma cell lines highly expressed MDK, indicating that it is a critical player in tumor development, which may henceforth represent an attractive therapeutic target. We showed that the knockdown of MDK expression by siRNA led to a marked and significant decrease in neuroblastoma (IGR-N91 and SH-SY5Y) cell proliferation in vitro. Using a new strategy, we then evaluated the antitumor effect of a truncated MDK protein, lacking the C-terminal 81-121 portion of the molecule (MDKD81-121), 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 MDKD81-121 selectively inhibited MDK-dependent tumor cells and was able to strongly reduce endothelial cell proliferation and migration and to induce ER stress-mediated apoptosis. We then investigated the effects of MDKD81-121 in vivo using electrotransfer of a plasmid encoding a secretable form of MDKD81-121 into tibialis cranialis muscles of nude mice. We showed that MDKD81-121 dramatically inhibited (up to 91%) tumor development and growth. This inhibition was correlated with the detection of the MDKD81-121 molecule in plasma and the suppression of intratumor neovascularization. Our findings demonstrate that MDK inhibition is a tractable therapeutic target for this lethal pediatric malignancy.

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Section: Plasmid Dnamentioning

confidence: 99%

Section: Cell Culture and Conditioned Mediummentioning

confidence: 99%

Section: Western Blot Analysis Elisa and Fishmentioning

confidence: 99%

Section: Western Blot Analysis Elisa and Fishmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Midkine Lacking Its Last 40 Amino Acids Acts on Endothelial and Neuroblastoma Tumor Cells and Inhibits Tumor Development

Dianat

Viet

Gobbo

et al. 2015

Molecular Cancer Therapeutics

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Antitumorigenic effects of a mutant of the heparin affin regulatory peptide on the U87 MG glioblastoma cell line

Santos

Karaky

Renoir

et al. 2010

Intl Journal of Cancer

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Glioblastoma is the most common primary brain tumor in human adults. Since existing treatments are not effective enough, novel therapeutic targets must be sought. The heparin-binding growth factor, heparin affin regulatory peptide (HARP), also known as pleiotrophin (PTN), could potentially represent such a target. We have previously shown that a mutant protein, HARPD111-136, which lacks HARP's C-terminal 26 amino acids, acts as a dominant negative HARP effector by heterodimerizing with the wild-type growth factor. The aim of our study was to evaluate the potential inhibitory activity of HARPD111-136 on the U87 MG human glioblastoma cell line. By overexpressing the truncated form of HARP in stably established clones of U87 MG cells, we observed an inhibition of proliferation under both anchorage-dependent and anchorage-independent conditions. We confirmed these results in an in vivo subcutaneous tumor xenograft model. In addition, we found that HARPD111-136 inhibited cell proliferation in a paracrine manner. Analysis of key cellular pathways revealed a decrease of cell adhesion in U87 MG cells that overexpressed the mutant protein, which could explain this inhibitory effect. A replication-defective adenovirus model that encoded HARPD111-136 supported a putative antiproliferative role for the truncated protein in vitro and in vivo. Interestingly, HARPD111-136 was also able to abolish angiogenic activity in HUVEC proliferation and in a Matrigel plug assay. These results demonstrate that considering its antiproliferative and angiostatic effects, HARPD111-136 could be of great interest when used in conjunction with standard treatments.Glioblastoma is the most common primary brain tumor in adult humans, with a median survival time of 12 months from the time of diagnosis, despite using standard therapeutic approaches (surgery, radiation and chemotherapy). A proper understanding of the principal pathways implicated in glioblastoma progression is needed to identify potential therapeutic targets. The heparin-binding growth factor, heparin affin regulatory peptide (HARP), also known as pleiotrophin (PTN) and heparin-binding growth-associated molecule (HB-GAM), could represent 1 such potential target.HARP is a 136-amino acid secreted polypeptide that forms with the protein midkine (MK), a 2-member family of heparin-binding growth factors. HARP is a developmentally regulated cytokine that is highly expressed in the embryonic nervous system, but its expression drops markedly after the perinatal phase and during adulthood. However, HARP expression may be deregulated, and HARP has been shown to be overexpressed in various tumor cell lines and primary human tumors, 1-3 including glioblastomas. In vitro, HARP displays a variety of biological activities including mitogenesis, angiogenesis, neurite outgrowth and cell migration. 4 Transforming activity has also been demonstrated for HARP when it is overexpressed in nontumor cell lines, such as NIH 3T3. 5Two transmembrane proteins with intracellular catalytic domains have been descri...

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A peptide corresponding to the C-terminal region of pleiotrophin inhibits angiogenesis in vivo and in vitro

et al. 2011

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Pleiotrophin (PTN) is a heparin-binding growth factor that plays a significant role in tumor growth and angiogenesis. We have previously shown that in order for PTN to induce migration of endothelial cells, binding to both α(ν) β(3) integrin and its receptor protein tyrosine phosphatase beta/zeta (RPTPβ/ζ) is required. In the present study we show that a synthetic peptide corresponding to the last 25 amino acids of the C-terminal region of PTN (PTN(112-136) ) inhibited angiogenesis in the in vivo chicken embryo chorioallantoic membrane (CAM) assay and PTN-induced migration and tube formation of human endothelial cells in vitro. PTN(112-136) inhibited binding of PTN to α(ν) β(3) integrin, and as shown by surface plasmon resonance (SPR) measurements, specifically interacted with the specificity loop of the extracellular domain of β(3) . Moreover, it abolished PTN-induced FAK Y397 phosphorylation, similarly to the effect of a neutralizing α(ν) β(3) -selective antibody. PTN(112-136) did not affect binding of PTN to RPTPβ/ζ in endothelial cells and induced β(3) Y773 phosphorylation and ERK1/2 activation to a similar extent with PTN. This effect was inhibited by down-regulation of RPTPβ/ζ by siRNA or by c-src inhibition, suggesting that PTN(112-136) may interact with RPTPβ/ζ. NMR spectroscopy studies showed that PTN(112-136) was characterized by conformational flexibility and absence of any element of secondary structure at room temperature, although the biologically active peptide segment 123-132 may adopt a defined structure at lower temperature. Collectively, our data suggest that although PTN(112-136) induces some of the signaling pathways triggered by PTN, it inhibits PTN-induced angiogenic activities through inhibition of PTN binding to α(ν) β(3) integrin.

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

Cited by 23 publications

References 39 publications

Midkine Lacking Its Last 40 Amino Acids Acts on Endothelial and Neuroblastoma Tumor Cells and Inhibits Tumor Development

Midkine Lacking Its Last 40 Amino Acids Acts on Endothelial and Neuroblastoma Tumor Cells and Inhibits Tumor Development

Antitumorigenic effects of a mutant of the heparin affin regulatory peptide on the U87 MG glioblastoma cell line

A peptide corresponding to the C-terminal region of pleiotrophin inhibits angiogenesis in vivo and in vitro

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