Tenascin-C Aptamers Are Generated Using Tumor Cells and Purified Protein

Hicke, Brian J.; Marion, Christopher; Chang, Ying-Fon; Gould, Ty; Lynott, C K; Parma, David H.; Schmidt, Paul G.; Warren, S.

doi:10.1074/jbc.m104651200

Cited by 306 publications

(211 citation statements)

References 51 publications

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“…A pool of RNA aptamers significantly inhibited S. enterica serovar Typhi invasion of THP-1 cells. Based on previous reports, an obvious improvement in aptamer-protein binding was usually apparent after five rounds of amplification (6,12). In this study, we decided to examine the biological activities of aptamers after the fifth, seventh, and eighth rounds of selection.…”

Section: Resultsmentioning

confidence: 99%

“…Bacterial self-association is an important virulence trait in enterotoxigenic strains of Escherichia coli and in Vibrio cholerae (1). These data indicated that the structural protein PilS of the type IVB pili might play important roles in the pathogenesis of S. enterica serovar Typhi in humans.The SELEX (systematic evolution of ligands by exponential enrichment) method (28) is an oligonucleotide-based combinatorial library selection procedure that has been used extensively to isolate ligands (aptamers) that bind to proteins (3,6,9,22,32), cell surface epitopes (19,21), and other targets (4,12,13,17). Although in recent years SELEX has become increasingly important in the study of functions of proteins, as well as in the fields of drug discovery and identification of antagonists against many functional proteins, this in vitro selection strategy to generate inhibitors of the functions of bacterial proteins remains underutilized.…”

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confidence: 99%

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Aptamers That Preferentially Bind Type IVB Pili and Inhibit Human Monocytic-Cell Invasion by Salmonella enterica Serovar Typhi

Pan¹,

Zhang

Wu³

et al. 2005

Antimicrob Agents Chemother

109

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Salmonella enterica serovar Typhi is an important pathogen exclusively for humans and causes typhoid or enteric fever. It has been shown that type IVB pili, encoded by the S. enterica serovar Typhi pil operon located in Salmonella pathogenicity island 7, are important in the pathogenic process. In this study, by using both an adhesion-invasion assay and fluorescence quantitative PCR analysis, we demonstrated that the entry of type IVB piliated S. enterica serovar Typhi A21-6 (pil ؉ Km r ) into human THP-1 monocytic cells was greater than that of a nonpiliated S. enterica serovar Typhi pilS::Km r (pil mutant) strain. We have applied a systematic evolution of ligands by exponential enrichment approach to select oligonucleotides (aptamers) as ligands that specifically bind to type IVB pili. Using this approach, we identified a high-affinity single-stranded RNA aptamer (S-PS 8.4 ) as a type IVB pilus-specific ligand and further found that the selected aptamer (S-PS 8.4 ) could significantly inhibit the entry of the piliated strain (but not that of the nonpiliated strain) into human THP-1 cells. The binding affinities between aptamers and pre-PilS (structural protein of type IVB pili) were determined by nitrocellulose filter-binding assays, and the K d value was determined to be 8.56 nM for the S-PS 8.4 aptamer alone. As an example of an aptamer against type IVB pili of S. enterica serovar Typhi, the aptamer S-PS 8.4 can serve as a tool for analysis of bacterial type IVB pilus-host cell interactions and may yield information for the development of putative new drugs against S. enterica serovar Typhi bacterial infections, useful both in prevention of infection and in therapeutic treatment.Of the more than 2,300 closely related Salmonella serovars identified, Salmonella enterica serovar Typhi is an important pathogen exclusively for humans and can be transmitted through contaminated food and water. It causes typhoid or enteric fever, which is a serious public health problem in developing countries. The genome of S. enterica serovar Typhi contains three large inserts (pathogenicity islands) (11), relative to the chromosome of Salmonella enterica serovar Typhimurium, which is normally noninvasive for humans. The type IVB pil operon of S. enterica serovar Typhi is located in Salmonella pathogenicity island 7 (18) and contains a pilS gene encoding the structural pilin (36, 37). It has been demonstrated that a pilS mutant of S. enterica serovar Typhi exhibited muchreduced adhesion to and invasion of human epithelial gastrointestinal cells in vitro and that purified soluble pre-PilS protein, retaining the signal sequence normally cleaved when the protein is excreted to form insoluble pili based on polymerized PilS, inhibited bacterial invasion (37). The structure of the N-terminally truncated type IVB structural pilin from S. enterica serovar Typhi was determined by nuclear magnetic resonance analysis (34). Type IVB pili, composed largely of polymerized PilS protein, also mediate bacterial self-association, but only when the...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Aptamers That Preferentially Bind Type IVB Pili and Inhibit Human Monocytic-Cell Invasion by Salmonella enterica Serovar Typhi

Pan¹,

Zhang

Wu³

et al. 2005

Antimicrob Agents Chemother

109

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“…43 Aptamers are stable, can be designed with desired length and specific binding affinities, and can be linked to other molecules such as cytokines. Using the SELEX technology on tenascin-C expressing U251 glioblastoma cells or on purified tenascin-C the aptamer "TTA1" was generated that showed high specificity and selectivity for human tenascin-C. 44 Further modifications improved biodistribution and led to the design of radionuclide labeled 99 mTc-TTA1. Upon intravenous injection into tumor bearing nude mice 99 mTc-TTA1 specifically had accumulated in the GBM xenograft tumor, 45 as well as in tumors induced by human colon, breast and rhabdomyosarcoma cells.…”

Section: Potential Use Of F16 For Radionuclide Therapymentioning

confidence: 99%

Tenascin-C: Exploitation and collateral damage in cancer management

Spenlé

Saupe

Midwood³

et al. 2015

Cell Adhesion & Migration

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“…Because shorter oligonucleotides are easier and more cost effectively produced, we truncated the best binding aptamer 157-B4 (47-mer) further by replacing nucleotides with a hexaethyleneglycol linker (26). The sequence variability of the AU-rich region was the most promising target to introduce the linker (Fig.…”

Section: Optimization Of Aptamer 157-b4 By Site-directed Modificationmentioning

confidence: 99%

An l -RNA-based aquaretic agent that inhibits vasopressin in vivo

Purschke

Eulberg

Buchner

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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A class of diuretic͞aquaretic agents based on mirror-image oligonucleotides (so-called Spiegelmers) has been identified. These molecules directly bind and inhibit the neuropeptide vasopressin (AVP). AVP is the major regulatory component of body fluid homeostasis mediated through binding to the renal V2 receptor. Elevated plasma levels of AVP are implicated in several pathological conditions, mainly cardiovascular diseases. In congestive heart failure, AVP is part of a neuroendocrine imbalance that is responsible for progressive worsening of the disease. Employing in vitro selection techniques, RNA aptamers that bind to the unnatural D-configuration of AVP were isolated. The best aptamer displayed an affinity to D-AVP of Ϸ560 pM at 37°C. The corresponding Spiegelmer, a 38-mer mirror-image oligonucleotide (L-RNA) termed NOX-F37, inhibits vasopressin-dependent activation of V1a as well as V2 receptors with IC50 values of 6.1 nM and 1 nM, respectively. NOX-F37 administered to healthy rats effectively neutralized AVP and increased diuresis dose-dependently for 24 h. The mode of action was strictly aquaretic, i.e., the increase in urine volume was not accompanied by an increase in electrolytes. These results clearly prove the in vivo efficacy of NOX-F37 and points out its potential as a drug in the treatment of diseases that are associated with body fluid overload.aptamer ͉ congestive heart failure ͉ diureses ͉ water homeostasis ͉ Spiegelmer T he cyclic nonapeptide vasopressin (AVP) is a neurohormone that is synthesized in the hypothalamus and stored in the hypophysis from where it is released into the circulation (1). AVP is also known as antidiuretic hormone because its main peripheral function is the maintenance of volume and osmolality of body fluids by regulating the free water reabsorption in the kidneys. Under physiological concentrations, AVP acts vasoconstrictive. AVP's actions are mediated through specific G proteincoupled receptors (2). The V 2 receptor, mainly expressed on cells of the basolateral membrane of the collecting duct in the kidneys, mediates the antidiuretic effect via enhanced cAMP levels (3). As a result, aquaporin channels are inserted into the cytoplasma membranes, and the transcription of aquaporin mRNA is induced. The V 1a receptor, mainly expressed on vascular smooth muscle cells and on cells of the myocardium, exerts vasoconstrictive effects through the second messengers inositoltrisphosphate and diacylglycerol. Subsequently, intracellular Ca 2ϩ is mobilized from the endoplasmic reticulum, and Ca 2ϩ channels as well as Na ϩ ͞H ϩ exchangers are activated (4). In addition to vasoactive effects, binding of AVP to the V 1a receptor also stimulates cell growth and proliferation in vascular smooth muscle cells through activation of the mitogen-activated protein kinase pathway (5).AVP release into the circulation is tightly regulated by osmoreceptors in the hypothalamus and baroreceptors in the heart and large arteries. Whereas osmoreceptors respond to small changes (1%) in osmolality, barorec...

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Tenascin-C Aptamers Are Generated Using Tumor Cells and Purified Protein

Cited by 306 publications

References 51 publications

Aptamers That Preferentially Bind Type IVB Pili and Inhibit Human Monocytic-Cell Invasion by Salmonella enterica Serovar Typhi

Aptamers That Preferentially Bind Type IVB Pili and Inhibit Human Monocytic-Cell Invasion by Salmonella enterica Serovar Typhi

Tenascin-C: Exploitation and collateral damage in cancer management

An l -RNA-based aquaretic agent that inhibits vasopressin in vivo

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