A new peptide ligand for targeting human carbonic anhydrase IX, identified through the phage display technology.

<h4>Unlabelled</h4>Carbonic anhydrase IX (CAIX) is a transmembrane enzyme found to be overexpressed in various tumors and associated with tumor hypoxia. Ligands binding this target may be used to visualize hypoxia, tumor manifestation or treat tumors by endoradiotherapy.<h4>Methods...

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Autores principales: Vasileios Askoxylakis, Regine Garcia-Boy, Shoaib Rana, Susanne Krämer, Ulrike Hebling, Walter Mier, Annette Altmann, Annette Markert, Jürgen Debus, Uwe Haberkorn
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Publicado: Public Library of Science (PLoS) 2010
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spelling oai:doaj.org-article:11895a8dce63466f99a24b4d221851d32021-11-18T07:00:52ZA new peptide ligand for targeting human carbonic anhydrase IX, identified through the phage display technology.1932-620310.1371/journal.pone.0015962https://doaj.org/article/11895a8dce63466f99a24b4d221851d32010-12-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21209841/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Unlabelled</h4>Carbonic anhydrase IX (CAIX) is a transmembrane enzyme found to be overexpressed in various tumors and associated with tumor hypoxia. Ligands binding this target may be used to visualize hypoxia, tumor manifestation or treat tumors by endoradiotherapy.<h4>Methods</h4>Phage display was performed with a 12 amino acid phage display library by panning against a recombinant extracellular domain of human carbonic anhydrase IX. The identified peptide CaIX-P1 was chemically synthesized and tested in vitro on various cell lines and in vivo in Balb/c nu/nu mice carrying subcutaneously transplanted tumors. Binding, kinetic and competition studies were performed on the CAIX positive human renal cell carcinoma cell line SKRC 52, the CAIX negative human renal cell carcinoma cell line CaKi 2, the human colorectal carcinoma cell line HCT 116 and on human umbilical vein endothelial cells (HUVEC). Organ distribution studies were carried out in mice, carrying SKRC 52 tumors. RNA expression of CAIX in HCT 116 and HUVEC cells was investigated by quantitative real time PCR.<h4>Results</h4>In vitro binding experiments of (125)I-labeled-CaIX-P1 revealed an increased uptake of the radioligand in the CAIX positive renal cell carcinoma cell line SKRC 52. Binding of the radioligand in the colorectal carcinoma cell line HCT 116 increased with increasing cell density and correlated with the mRNA expression of CAIX. Radioligand uptake was inhibited up to 90% by the unlabeled CaIX-P1 peptide, but not by the negative control peptide octreotide at the same concentration. No binding was demonstrated in CAIX negative CaKi 2 and HUVEC cells. Organ distribution studies revealed a higher accumulation in SKRC 52 tumors than in heart, spleen, liver, muscle, intestinum and brain, but a lower uptake compared to blood and kidney.<h4>Conclusions</h4>These data indicate that CaIX-P1 is a promising candidate for the development of new ligands targeting human carbonic anhydrase IX.Vasileios AskoxylakisRegine Garcia-BoyShoaib RanaSusanne KrämerUlrike HeblingWalter MierAnnette AltmannAnnette MarkertJürgen DebusUwe HaberkornPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 12, p e15962 (2010)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Vasileios Askoxylakis
Regine Garcia-Boy
Shoaib Rana
Susanne Krämer
Ulrike Hebling
Walter Mier
Annette Altmann
Annette Markert
Jürgen Debus
Uwe Haberkorn
A new peptide ligand for targeting human carbonic anhydrase IX, identified through the phage display technology.
description <h4>Unlabelled</h4>Carbonic anhydrase IX (CAIX) is a transmembrane enzyme found to be overexpressed in various tumors and associated with tumor hypoxia. Ligands binding this target may be used to visualize hypoxia, tumor manifestation or treat tumors by endoradiotherapy.<h4>Methods</h4>Phage display was performed with a 12 amino acid phage display library by panning against a recombinant extracellular domain of human carbonic anhydrase IX. The identified peptide CaIX-P1 was chemically synthesized and tested in vitro on various cell lines and in vivo in Balb/c nu/nu mice carrying subcutaneously transplanted tumors. Binding, kinetic and competition studies were performed on the CAIX positive human renal cell carcinoma cell line SKRC 52, the CAIX negative human renal cell carcinoma cell line CaKi 2, the human colorectal carcinoma cell line HCT 116 and on human umbilical vein endothelial cells (HUVEC). Organ distribution studies were carried out in mice, carrying SKRC 52 tumors. RNA expression of CAIX in HCT 116 and HUVEC cells was investigated by quantitative real time PCR.<h4>Results</h4>In vitro binding experiments of (125)I-labeled-CaIX-P1 revealed an increased uptake of the radioligand in the CAIX positive renal cell carcinoma cell line SKRC 52. Binding of the radioligand in the colorectal carcinoma cell line HCT 116 increased with increasing cell density and correlated with the mRNA expression of CAIX. Radioligand uptake was inhibited up to 90% by the unlabeled CaIX-P1 peptide, but not by the negative control peptide octreotide at the same concentration. No binding was demonstrated in CAIX negative CaKi 2 and HUVEC cells. Organ distribution studies revealed a higher accumulation in SKRC 52 tumors than in heart, spleen, liver, muscle, intestinum and brain, but a lower uptake compared to blood and kidney.<h4>Conclusions</h4>These data indicate that CaIX-P1 is a promising candidate for the development of new ligands targeting human carbonic anhydrase IX.
format article
author Vasileios Askoxylakis
Regine Garcia-Boy
Shoaib Rana
Susanne Krämer
Ulrike Hebling
Walter Mier
Annette Altmann
Annette Markert
Jürgen Debus
Uwe Haberkorn
author_facet Vasileios Askoxylakis
Regine Garcia-Boy
Shoaib Rana
Susanne Krämer
Ulrike Hebling
Walter Mier
Annette Altmann
Annette Markert
Jürgen Debus
Uwe Haberkorn
author_sort Vasileios Askoxylakis
title A new peptide ligand for targeting human carbonic anhydrase IX, identified through the phage display technology.
title_short A new peptide ligand for targeting human carbonic anhydrase IX, identified through the phage display technology.
title_full A new peptide ligand for targeting human carbonic anhydrase IX, identified through the phage display technology.
title_fullStr A new peptide ligand for targeting human carbonic anhydrase IX, identified through the phage display technology.
title_full_unstemmed A new peptide ligand for targeting human carbonic anhydrase IX, identified through the phage display technology.
title_sort new peptide ligand for targeting human carbonic anhydrase ix, identified through the phage display technology.
publisher Public Library of Science (PLoS)
publishDate 2010
url https://doaj.org/article/11895a8dce63466f99a24b4d221851d3
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