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

Abstract

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.

Structural studies reveal an important role for the pleiotrophin C‐terminus in mediating interactions with chondroitin sulfate

“…inal tail can inhibit PTN activity [38]. However, previous studies of midkine and PTN showed the C-terminal tails of the respective proteins are not necessary for binding heparin [26,28].…”

“…The C‐terminal tail of PTN is necessary for PTPRZ signaling , and peptides derived from the C‐terminal tail can inhibit PTN activity . However, previous studies of midkine and PTN showed the C‐terminal tails of the respective proteins are not necessary for binding heparin .…”

Implications of pleiotrophin in human PC3 prostate cancer cell growth in vivo

“…Similarly, a peptide that corresponds to the carboxy‐terminal domain of PTN and binds to RPTPβ/ζ, inhibits prostate cancer cell response to PTN in vitro and PC3 tumor growth in vivo , although in the latter case it has not been shown whether its effect was solely due to endogenous PTN inhibition through RPTPβ/ζ and not interference with other implicated pathways. We have previously shown that a similar peptide inhibits the stimulatory effect of PTN on endothelial cell migration through inhibition of PTN interaction with α ν β 3 integrin . Integrins, and especially α ν β 3 , play a role in prostate cancer cell metastasis to bone .…”

Small Molecules that Rescue Multiple Phenotypic Aberrations in an iPSC-Derived Neuron Model of CLN3 Disease

“…The Matrigel tube formation assay was performed as previously described (70). Wells of a 96-well plate were coated with 40 µL Growth Factor-Reduced (RGF) basement membrane extract (Trevigen) and incubated for 20 minutes at 37 °C to polymerize.…”