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J. Biol. Chem., Vol. 276, Issue 19, 16464-16468, May 11, 2001
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From the Tumor microvessels differ in structure and
metabolic function from normal vasculature, and neoangiogenesis is
associated with quantitative and qualitative changes in expression of
endothelial proteins. Such molecules could serve as molecular addresses
differentiating the tumor vasculature from those of the normal brain.
We have applied Systematic Evolution of
Ligands by EXponential enrichment (SELEX)
against transformed endothelial cells as a complex target to select
single-stranded DNA-ligands (aptamers) that function as
histological markers to detect microvessels of rat experimental glioma,
a fatal brain tumor that is highly vascularized. Both the SELEX
selection procedure as well as subsequent deconvolution-SELEX were
analyzed by fluorescence based methods (flow cytometry and fluorescence
microscopy). Of 25 aptamers analyzed, one aptamer was selected that
selectively bound microvessels of rat brain glioblastoma but not the
vasculature of the normal rat brain including peritumoral areas. The
molecular target protein of aptamer III.1 was isolated from endothelial
cells by ligand-mediated magnetic DNA affinity purification. This
protein was identified by mass spectrometry as rat homologue of mouse
pigpen, a not widely known endothelial protein the expression of which
parallels the transition from quiescent to angiogenic phenotypes
in vitro. Because neoangiogenesis, the formation of new
blood vessels, is a key feature of tumor development, the presented
aptamer can be used as a probe to analyze pathological angiogenesis of
glioblastoma. The presented data show that pigpen is highly expressed
in tumor microvessels of experimental rat brain glioblastoma and may
play an important role in warranting blood supply, thus growth of brain tumors.
Systematic Evolution of a DNA Aptamer Binding to Rat Brain Tumor
Microvessels
SELECTIVE TARGETING OF ENDOTHELIAL REGULATORY PROTEIN
PIGPEN*
§,
, and
Institute of Brain Research, University of
Tuebingen, Calwer Strasse 3, D-72076 Tuebingen, Germany, the
¶ Institute for Cell Biology, Department of Immunology, University
of Tuebingen, Auf der Morgenstelle 15, D-72076 Tuebingen, Germany, and
the Institute for Cell Biology, Department of Molecular Biology,
University of Tuebingen, Auf der Morgenstelle 15, D-72076 Tuebingen, Germany
*
This work was supported by the Wilhelm Sander Foundation.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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