Previously developed Asn-Gly-Arg(NGR) peptide-modified multifunctional poly(ethyleneimine)–poly(ethylene glycol)(PEI–PEG)-based nanoparticles(TPIC) have been considered to be promising carriers for the co-delivery o...Previously developed Asn-Gly-Arg(NGR) peptide-modified multifunctional poly(ethyleneimine)–poly(ethylene glycol)(PEI–PEG)-based nanoparticles(TPIC) have been considered to be promising carriers for the co-delivery of DNA and doxorubicin(DOX). As a continued effort, the aim of the present study was to further evaluate the interaction between TPIC and human umbilical vein endothelial cells(HUVEC) to better understand the cellular entry mechanism. In the present investigation,experiments relevant to co-localization, endocytosis inhibitors and factors influencing the internalization were performed. Without any treatment, there was no co-localization between aminopeptidase N/CD13(APN/CD13) and caveolin 1(CAV1). However, co-localization between CD13 and CAV1 was observed when cells were incubated with an anti-CD13 antibody or TPIC. As compared with antibody treatment,TPIC accelerated the speed and enhanced the degree of co-localization. TPIC entered HUVEC not only together with CD13 but also together with CAV1. However, this internalization was not dependent on the enzyme activity of CD13 but could be inhibited by methyl-β-eyclodextfin(MβCD), further identifying the involvement of caveolae-mediated endocytosis(CvME). This conclusion was also verified by endocytosis inhibitor experiments.展开更多
In the present study, we prepared novel NGR-modified PEG-PLGA polymeric micelles containing paclitaxel （NGR- PM-PTX） in order to evaluate their potential targeting to aminopeptidase N receptors expressed on tumor en...In the present study, we prepared novel NGR-modified PEG-PLGA polymeric micelles containing paclitaxel （NGR- PM-PTX） in order to evaluate their potential targeting to aminopeptidase N receptors expressed on tumor endothelial cells and the tumor cell surface and its anti-tumor activity in vitro and in vivo. NGR-PM-PTX was prepared by thin-film hydration method. The in vitro targeting characteristics of NGR-modified PM on HUVEC （human umbilical vein endothelial cells）, HT1080 （human fibrosarcoma cells） and MCF-7 （human breast adenocarcinoma cells） were then investigated. The anti-tumor activity of NGR-PM-PTX was evaluated in HT1080 tumor-bearing mice in vivo. The targeting activity of the NGR-modified PM was demonstrated by flow cytometry and confocal microscopy in vitro. NGR-PM-PTX also produced marked anti-tumor activity to HTI080 tumor-beating mice in vivo.展开更多
Colloidal gold （Au）, a well-tolerated several applications in nanomedicine. nanomaterial, is currently exploited for We show that gold nanoparticles tagged with a novel tumor-homing peptide containing Asn-Gly-Arg （...Colloidal gold （Au）, a well-tolerated several applications in nanomedicine. nanomaterial, is currently exploited for We show that gold nanoparticles tagged with a novel tumor-homing peptide containing Asn-Gly-Arg （NGR）, a ligand of CD13 expressed by the tumor neovasculature, can be exploited as carriers for cytokine delivery to tumors. Biochemical and functional studies showed that the NGR molecular scaffoldflinker used for gold functionalization is critical for CD13 recognition. Using fibrosarcorna-bearing mice, NGR-tagged nanodrugs could deliver extremely low, yet pharmacologically active doses of tumor necrosis factor （TNF）, an anticancer cytokine, to tumors with no evidence of toxicity. Mechanistic studies confirmed that CD13 targeting was a primary mechanism of drug delivery and excluded a major role of integrin targeting consequent to NGR deamidation, a degradation reaction that generates the isoAsp-Gly-Arg （isoDGR） integrin ligand. NGR-tagged gold nanoparticles can be used, in principle, as a novel platform for single- or multi-cytokine delivery to tumor endothelial cells for cancer therapy.展开更多
基金supported by the National Natural Science Foundation(No.81402867)
文摘Previously developed Asn-Gly-Arg(NGR) peptide-modified multifunctional poly(ethyleneimine)–poly(ethylene glycol)(PEI–PEG)-based nanoparticles(TPIC) have been considered to be promising carriers for the co-delivery of DNA and doxorubicin(DOX). As a continued effort, the aim of the present study was to further evaluate the interaction between TPIC and human umbilical vein endothelial cells(HUVEC) to better understand the cellular entry mechanism. In the present investigation,experiments relevant to co-localization, endocytosis inhibitors and factors influencing the internalization were performed. Without any treatment, there was no co-localization between aminopeptidase N/CD13(APN/CD13) and caveolin 1(CAV1). However, co-localization between CD13 and CAV1 was observed when cells were incubated with an anti-CD13 antibody or TPIC. As compared with antibody treatment,TPIC accelerated the speed and enhanced the degree of co-localization. TPIC entered HUVEC not only together with CD13 but also together with CAV1. However, this internalization was not dependent on the enzyme activity of CD13 but could be inhibited by methyl-β-eyclodextfin(MβCD), further identifying the involvement of caveolae-mediated endocytosis(CvME). This conclusion was also verified by endocytosis inhibitor experiments.
基金National Natural Science Foundation of China (Grant No.30873170)
文摘In the present study, we prepared novel NGR-modified PEG-PLGA polymeric micelles containing paclitaxel （NGR- PM-PTX） in order to evaluate their potential targeting to aminopeptidase N receptors expressed on tumor endothelial cells and the tumor cell surface and its anti-tumor activity in vitro and in vivo. NGR-PM-PTX was prepared by thin-film hydration method. The in vitro targeting characteristics of NGR-modified PM on HUVEC （human umbilical vein endothelial cells）, HT1080 （human fibrosarcoma cells） and MCF-7 （human breast adenocarcinoma cells） were then investigated. The anti-tumor activity of NGR-PM-PTX was evaluated in HT1080 tumor-bearing mice in vivo. The targeting activity of the NGR-modified PM was demonstrated by flow cytometry and confocal microscopy in vitro. NGR-PM-PTX also produced marked anti-tumor activity to HTI080 tumor-beating mice in vivo.
文摘Colloidal gold （Au）, a well-tolerated several applications in nanomedicine. nanomaterial, is currently exploited for We show that gold nanoparticles tagged with a novel tumor-homing peptide containing Asn-Gly-Arg （NGR）, a ligand of CD13 expressed by the tumor neovasculature, can be exploited as carriers for cytokine delivery to tumors. Biochemical and functional studies showed that the NGR molecular scaffoldflinker used for gold functionalization is critical for CD13 recognition. Using fibrosarcorna-bearing mice, NGR-tagged nanodrugs could deliver extremely low, yet pharmacologically active doses of tumor necrosis factor （TNF）, an anticancer cytokine, to tumors with no evidence of toxicity. Mechanistic studies confirmed that CD13 targeting was a primary mechanism of drug delivery and excluded a major role of integrin targeting consequent to NGR deamidation, a degradation reaction that generates the isoAsp-Gly-Arg （isoDGR） integrin ligand. NGR-tagged gold nanoparticles can be used, in principle, as a novel platform for single- or multi-cytokine delivery to tumor endothelial cells for cancer therapy.
