In vitro selection of DNA aptamers that block toxic effects of AGE on cultured retinal pericytes

Yuichiro Higashimoto, Sho ichi Yamagishi, Kazuo Nakamura, Takanori Matsui, Masayoshi Takeuchi, Masato Noguchi, Hiroyoshi Inoue

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Microvessels are composed of endothelial cells and pericytes. We have previously shown that advanced glycation end products (AGE) not only inhibit DNA synthesis but also induce apoptosis in cultured retinal pericytes, thereby being involved in pericyte loss, the earliest histopathological hallmark of diabetic retinopathy. Since pericytes play a central role in the maintenance of microvascular homeostasis in the retina, blockade of the harmful effects of AGE on retinal pericytes may become a novel therapeutic strategy for the treatment of diabetic retinopathy. In this study, we selected DNA aptamers directed against AGE in vitro and then examined their cytoprotective effects on AGE-exposed retinal pericytes. We identified 15 DNA aptamers directed against AGE-human serum albumin using combinatorial chemistry techniques in vitro. Structural analysis revealed that they had bulge-loop structures with cytosine-rich sequences. All of the aptamers, but not non-binding control aptamers, were found to inhibit the AGE-induced decrease in DNA synthesis as well as apoptotic cell death in pericytes. Among the selected aptamers, the clone 9 aptamer completely blocked the toxic effects of AGE, and its dissociation constant was 1 μmol/L. These results indicate that DNA aptamers are a useful tool for inhibiting the cytotoxic effects of AGE on cultured retinal pericytes. Our study suggests that blockade of the AGE effects by DNA aptamers may lead to a novel therapeutic strategy for the treatment of diabetic retinopathy.

Original languageEnglish
Pages (from-to)65-69
Number of pages5
JournalMicrovascular Research
Volume74
Issue number1
DOIs
Publication statusPublished - 2007 Jul
Externally publishedYes

Fingerprint

Nucleotide Aptamers
Pericytes
Advanced Glycosylation End Products
Poisons
Diabetic Retinopathy
Combinatorial Chemistry Techniques
In Vitro Techniques
Cytosine
DNA
Endothelial cells
Cell death
Microvessels
Structural analysis
Retina
Homeostasis
Cell Death
Endothelial Cells
Clone Cells
Maintenance
Apoptosis

Keywords

  • AGE
  • Diabetic retinopathy
  • DNA aptamer
  • Pericyte loss

ASJC Scopus subject areas

  • Biochemistry
  • Cardiology and Cardiovascular Medicine

Cite this

In vitro selection of DNA aptamers that block toxic effects of AGE on cultured retinal pericytes. / Higashimoto, Yuichiro; Yamagishi, Sho ichi; Nakamura, Kazuo; Matsui, Takanori; Takeuchi, Masayoshi; Noguchi, Masato; Inoue, Hiroyoshi.

In: Microvascular Research, Vol. 74, No. 1, 07.2007, p. 65-69.

Research output: Contribution to journalArticle

Higashimoto, Y, Yamagishi, SI, Nakamura, K, Matsui, T, Takeuchi, M, Noguchi, M & Inoue, H 2007, 'In vitro selection of DNA aptamers that block toxic effects of AGE on cultured retinal pericytes', Microvascular Research, vol. 74, no. 1, pp. 65-69. https://doi.org/10.1016/j.mvr.2007.04.001
Higashimoto Y, Yamagishi SI, Nakamura K, Matsui T, Takeuchi M, Noguchi M et al. In vitro selection of DNA aptamers that block toxic effects of AGE on cultured retinal pericytes. Microvascular Research. 2007 Jul;74(1):65-69. https://doi.org/10.1016/j.mvr.2007.04.001
Higashimoto, Yuichiro ; Yamagishi, Sho ichi ; Nakamura, Kazuo ; Matsui, Takanori ; Takeuchi, Masayoshi ; Noguchi, Masato ; Inoue, Hiroyoshi. / In vitro selection of DNA aptamers that block toxic effects of AGE on cultured retinal pericytes. In: Microvascular Research. 2007 ; Vol. 74, No. 1. pp. 65-69.
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