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      Oligonucleotide Aptamers: New Tools for Targeted Cancer Therapy

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          Abstract

          Aptamers are a class of small nucleic acid ligands that are composed of RNA or single-stranded DNA oligonucleotides and have high specificity and affinity for their targets. Similar to antibodies, aptamers interact with their targets by recognizing a specific three-dimensional structure and are thus termed “chemical antibodies.” In contrast to protein antibodies, aptamers offer unique chemical and biological characteristics based on their oligonucleotide properties. Hence, they are more suitable for the development of novel clinical applications. Aptamer technology has been widely investigated in various biomedical fields for biomarker discovery, in vitro diagnosis, in vivo imaging, and targeted therapy. This review will discuss the potential applications of aptamer technology as a new tool for targeted cancer therapy with emphasis on the development of aptamers that are able to specifically target cell surface biomarkers. Additionally, we will describe several approaches for the use of aptamers in targeted therapeutics, including aptamer-drug conjugation, aptamer-nanoparticle conjugation, aptamer-mediated targeted gene therapy, aptamer-mediated immunotherapy, and aptamer-mediated biotherapy.

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          Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase.

          L Gold, C Tuerk (1990)
          High-affinity nucleic acid ligands for a protein were isolated by a procedure that depends on alternate cycles of ligand selection from pools of variant sequences and amplification of the bound species. Multiple rounds exponentially enrich the population for the highest affinity species that can be clonally isolated and characterized. In particular one eight-base region of an RNA that interacts with the T4 DNA polymerase was chosen and randomized. Two different sequences were selected by this procedure from the calculated pool of 65,536 species. One is the wild-type sequence found in the bacteriophage mRNA; one is varied from wild type at four positions. The binding constants of these two RNA's to T4 DNA polymerase are equivalent. These protocols with minimal modification can yield high-affinity ligands for any protein that binds nucleic acids as part of its function; high-affinity ligands could conceivably be developed for any target molecule.
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            Development of DNA aptamers using Cell-SELEX.

            Kwame Sefah, Dihua Shangguan, Xiangling Xiong … (2010)
            In the past two decades, high-affinity nucleic acid aptamers have been developed for a wide variety of pure molecules and complex systems such as live cells. Conceptually, aptamers are developed by an evolutionary process, whereby, as selection progresses, sequences with a certain conformation capable of binding to the target of interest emerge and dominate the pool. This protocol, cell-SELEX (systematic evolution of ligands by exponential enrichment), is a method that can generate DNA aptamers that can bind specifically to a cell type of interest. Commonly, a cancer cell line is used as the target to generate aptamers that can differentiate that cell type from other cancers or normal cells. A single-stranded DNA (ssDNA) library pool is incubated with the target cells. Nonbinding sequences are washed off and bound sequences are recovered from the cells by heating cell-DNA complexes at 95 degrees C, followed by centrifugation. The recovered pool is incubated with the control cell line to filter out the sequences that bind to common molecules on both the target and the control, leading to the enrichment of specific binders to the target. Binding sequences are amplified by PCR using fluorescein isothiocyanate-labeled sense and biotin-labeled antisense primers. This is followed by removal of antisense strands to generate an ssDNA pool for subsequent rounds of selection. The enrichment of the selected pools is monitored by flow cytometry binding assays, with selected pools having increased fluorescence compared with the unselected DNA library. The procedure, from design of oligonucleotides to enrichment of the selected pools, takes approximately 3 months.
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              Aptamers and Their Biological Applications

              Kyung–Mi Song, Seonghwan Lee, Changill Ban (2012)
              Recently, aptamers have attracted the attention of many scientists, because they not only have all of the advantages of antibodies, but also have unique merits, such as thermal stability, low cost, and unlimited applications. In this review, we present the reasons why aptamers are known as alternatives to antibodies. Furthermore, several types of in vitro selection processes, including nitrocellulose membrane filtration, affinity chromatography, magnetic bead, and capillary electrophoresis-based selection methods, are explained in detail. We also introduce various applications of aptamers for the diagnosis of diseases and detection of small molecules. Numerous analytical techniques, such as electrochemical, colorimetric, optical, and mass-sensitive methods, can be utilized to detect targets, due to convenient modifications and the stability of aptamers. Finally, several medical and analytical applications of aptamers are presented. In summary, aptamers are promising materials for diverse areas, not just as alternatives to antibodies, but as the core components of medical and analytical equipment.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Mol Ther Nucleic Acids
                Journal ID (iso-abbrev): Mol Ther Nucleic Acids
                Title: Molecular Therapy. Nucleic Acids
                Publisher: Nature Publishing Group
                ISSN (Electronic): 2162-2531
                Publication date (Print): August 2014
                Publication date (Electronic): 05 August 2014
                Publication date PMC-release: 1 August 2014
                Volume: 3
                Issue: 8
                Page: e182
                Affiliations
                [1 ]Department of Pathology and Genomic Medicine, Houston Methodist Hospital , Houston, Texas, USA
                [2 ]Department of Immunology, Norman Bethune College of Medicine, Jilin University , Jilin, China
                [3 ]Sirnaomics Inc. , Gaithersburg, Maryland, USA
                [4 ]School of Biosciences and Bioengineering, South China University of Technology , Guangzhou, China
                Author notes
                [* ]Department of Pathology and Genomic Medicine, Houston Methodist Hospital, Houston, Texas, USA. E-mail: yzu@ 123456HoustonMethodist.org
                [* ]School of Biosciences and Bioengineering, South China University of Technology, Guangzhou, China. E-mail: went@ 123456scut.edu.cn
                Article
                Publisher Item ID: mtna201432
                DOI: 10.1038/mtna.2014.32
                PMC ID: 4221593
                PubMed ID: 25093706
                SO-VID: f8ce42ba-4125-4f74-8b4f-0ecfb82ae833
                Copyright © Copyright © 2014 American Society of Gene & Cell Therapy
                License:

                This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/

                History
                Date received : 12 April 2014
                Date accepted : 30 May 2014
                Categories
                Subject: Review

                ScienceOpen disciplines: Molecular medicine
                Keywords: cell surface biomarker,nanomedicine,oligonucleotide aptamer,selex,targeted cancer therapy
                Data availability:
                ScienceOpen disciplines: Molecular medicine
                Keywords: cell surface biomarker, nanomedicine, oligonucleotide aptamer, selex, targeted cancer therapy

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