Prevention of passively transferred experimental autoimmune myasthenia gravis by an in vitro selected RNA aptamer

Hwang, Byounghoon; Han, Kyungsook; Lee, Seong Wook

doi:10.1016/s0014-5793(03)00745-2

Cited by 35 publications

(17 citation statements)

References 26 publications

(37 reference statements)

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“…A great advantage of this technology is that it can select sequences capable of binding to a variety of ligands, including small molecules, inorganics, proteins, and cellular targets (30). Good progress toward clinical applications of aptamers is being made (21,(31)(32)(33)(34)(35). At the same time, oligonucleotides containing G-quartets (36), CpG sequences (37,38), and transcription factor decoys (39 -41) have recently generated much interest for their ability to modulate disease states and the immune system.…”

Section: Discussionmentioning

confidence: 99%

Construction, Stability, and Activity of Multivalent Circular Anticoagulant Aptamers

Giusto

King

2004

Journal of Biological Chemistry

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Here we describe the design and construction of multivalent circular DNA aptamers. Four aptameric binding motifs directed at blood-borne targets are used as a model set from which larger, multidomain aptamers are constructed in a straightforward manner. Intra-or intermolecular ligation of precursor oligonucleotides provides a stabilizing mechanism against degradation by the predominant exonuclease activity of blood products without the need for post-selection chemical modification. In many cases, circular DNA aptamer half-lives are extended beyond 10 h in serum and plasma, making such constructs viable for therapeutic and diagnostic applications. Duplexes and three-way junctions are used as scaffold architectures around which two, three, or four aptameric motifs can be arranged in a structurally defined manner, giving rise to improved binding characteristics through stability and avidity gains. Circular aptamers targeted against thrombin display improved anticoagulant potency with EC 50 values 2-3-fold better than those of the canonical GS-522 thrombin DNA aptamer. Intrinsic duplex regions provide an opportunity to incorporate additional transcription factor binding motifs, whereas ancillary loops can be used to provide further functionality. These anticoagulant aptamers provide a starting point for merging the principles of DNA nanotechnology with aptameric functions.Since the first reports on nucleic acid aptamers more than a decade ago (1-3), this class of molecules has developed steadily toward general therapeutic (4 -6) and diagnostic (5, 7-9) applications. The relative ease of aptamer discovery has allowed for selection against a wide range of targets (10) by processes that have recently been automated (11). Simultaneous advances in signaling and detection strategies have improved the outlook for aptamer arrays in proteomic and biosensor applications. Whereas the high affinity and specificity of aptamers toward their molecular targets make them well suited for these tasks, the susceptibility of natural nucleic acids to nucleolytic degradation is a serious hurdle for uses in biological fluids. Strategies to improve the nuclease stability of aptamers, as for antisense nucleic acids, have included the use of chemical modifications such as phosphorothioate, locked nucleic acid, 2Ј-Omethyl-and 2Ј-fluoro-nucleotides (12), as well as mirror-image Spiegelmers (13). Of most relevance for potential therapeutic applications, some common nucleic acid stabilization strategies have encountered problems associated with toxic degradation products (14). In addition, modification performed post-selection can alter the subtle binding interactions of the selected natural aptamer (15, 16).Circularization of natural aptamers is an attractive alternative to chemical modification for improving aptamer stability. With the majority of nucleic acid degradation activity arising from plasma exonucleases (17), modification of exposed termini often achieves a sufficient improvement in stability for use in vivo (12), and circular constru...

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Section: Discussionmentioning

confidence: 99%

Construction, Stability, and Activity of Multivalent Circular Anticoagulant Aptamers

Giusto

King

2004

Journal of Biological Chemistry

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“…Aptamers to extracellular targets could be used for the treatment of angiogenesis [18,38,172] , inflammation [175][176][177] , anticoagulation and thrombosis [6,81,173,174,178] as well as autoimmune diseases [179][180][181][182][183][184] . Such aptamers could be administered intravenously or subcutaneously.…”

Section: Aptamers To Extracellular Targetsmentioning

confidence: 99%

Characterisation of aptamers for therapeutic studies

Wochner¹,

Menger²,

Rimmele³

2007

Expert Opinion on Drug Discovery

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Nucleic acid aptamers, like monoclonal antibodies, bind to their target molecule with high affinity and specificity. Owing to their molecular recognition properties that are based on their three-dimensional structure, aptamers have been used as powerful tools in various fields, such as detection, separation or purification of target molecules. In addition, their intrinsic characteristics make aptamers excellent candidates for therapeutic applications. Due to their high specificity, stability, low toxicity and apparent non-immunogenicity, they provide viable alternatives to antibody- and small molecule-based therapeutics. Unlike antibodies, they can be generated against toxic, unstable or difficult to purify proteins. Furthermore, they can be chemically derivatised to extend their bioavailability and lifetimes in biological fluids, and they even offer the possibility for fast and efficient regulation. Similar to other therapeutic nucleic acids, such as antisense oligonucleotides, ribozymes or siRNA, problems with cellular uptake and susceptibility to nucleolytic attack have to be overcome for their successful application. With the first aptamer-based drug having entered the market and a few in Phase II trials, the road has been paved for more to follow. In this review, the requirements and properties of therapeutic aptamers are discussed and the recent progress in aptamer research towards drug development is highlighted.

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“…Such short RNA ligands, termed RNA aptamers, have been identified from a random RNA library to bind several proteins including non-RNA-binding proteins with high affinity and specificity using in vitro selection techniques, called systemic evolution of ligands by exponential enrichment (SELEX; Ellington and Szostak 1990;Tuerk and Gold 1990). Several RNA aptamers have been successfully evaluated even in animal disease models (Sullenger and Gilboa 2002;Hwang et al 2003).…”

Section: Introductionmentioning

confidence: 99%

Isolation of specific and high-affinity RNA aptamers against NS3 helicase domain of hepatitis C virus

Hwang¹,

Cho²,

Yeo³

et al. 2004

RNA

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Hepatitis C virus (HCV)-encoded nonstructural protein 3 (NS3) possesses protease, NTPase, and helicase activities, which are considered essential for viral proliferation. Thus, HCV NS3 is a good putative therapeutic target protein for the development of anti-HCV agents. In this study, we isolated specific RNA aptamers to the helicase domain of HCV NS3 from a combinatorial RNA library with 40-nucleotide random sequences using in vitro selection techniques. The isolated RNAs were observed to very avidly bind the HCV helicase with an apparent K d of 990 pM in contrast to original pool RNAs with a K d of >1 µM. These RNA ligands appear to impede binding of substrate RNA to the HCV helicase and can act as potent decoys to competitively inhibit helicase activity with high efficiency compared with poly(U) or tRNA. The minimal binding domain of the ligands was determined to evaluate the structural features of the isolated RNA molecules. Interestingly, part of binding motif of the RNA aptamers consists of similar secondary structure to the 3-end of HCV negative-strand RNA. Moreover, intracellular NS3 protein can be specifically detected in situ with the RNA aptamers, indicating that the selected RNAs are very specific to the HCV NS3 helicase. Furthermore, the RNA aptamers partially inhibited RNA synthesis of HCV subgenomic replicon in Huh-7 hepatoma cell lines. These results suggest that the RNA aptamers selected in vitro could be useful not only as therapeutic and diagnostic agents of HCV infection but also as a powerful tool for the study of HCV helicase mechanism.

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Prevention of passively transferred experimental autoimmune myasthenia gravis by an in vitro selected RNA aptamer

Cited by 35 publications

References 26 publications

Construction, Stability, and Activity of Multivalent Circular Anticoagulant Aptamers

Construction, Stability, and Activity of Multivalent Circular Anticoagulant Aptamers

Characterisation of aptamers for therapeutic studies

Isolation of specific and high-affinity RNA aptamers against NS3 helicase domain of hepatitis C virus

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