Related Experiment Videos
The generation and characterization of antagonist RNA aptamers to human oncostatin M
1Molecular Discovery Department, Glaxo Wellcome Research and Development, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY, UK. adr7003@glaxowellcome.co.uk
Abstract:
Oncostatin M (OSM) is a multifunctional member of the interleukin-6 cytokine family. OSM has been implicated as a powerful proinflammatory mediator and may represent a potentially important, novel therapeutic opportunity for treatment of established rheumatoid arthritis. To further investigate the role of OSM in inflammatory disorders, we have isolated a series of RNA aptamers that bind specifically to human OSM. The highest affinity aptamer, designated ADR58, has been characterized in a series of in vitro and cell based assays. ADR58 has an affinity of 7 nm for human OSM, and it can antagonize OSM binding to the gp130 receptor and specifically antagonize OSM mediated signaling. The aptamer has been truncated in length to 33 bases, all pyrimidine positions are substituted with 2' fluorine, and 14 of 18 purine positions have been substituted with 2' O-methyl to increase stability toward nucleases. This truncated, modified form of ADR58 retains complete affinity and functional activity for OSM. This aptamer may be used as a tool to further investigate the role of OSM in inflammatory disorders and may also have role as a therapeutic agent.
Insights
Researchers developed a novel RNA aptamer, ADR58, that specifically targets Oncostatin M (OSM), a key inflammatory mediator. This aptamer effectively blocks OSM signaling, offering potential for rheumatoid arthritis treatment and research.
Area of Science:
- Biochemistry
- Molecular Biology
- Immunology
Background:
- Oncostatin M (OSM), a cytokine in the interleukin-6 family, is a potent pro-inflammatory mediator.
- OSM plays a significant role in inflammatory disorders, including rheumatoid arthritis, presenting a therapeutic target.
Purpose of the Study:
- To investigate the role of OSM in inflammatory disorders.
- To develop novel RNA aptamers that specifically bind to and inhibit human OSM.
- To characterize the highest affinity aptamer for its potential therapeutic applications.
Main Methods:
- Isolation and characterization of RNA aptamers targeting human OSM.
- Affinity determination using in vitro and cell-based assays.
- Assessment of aptamer's ability to antagonize OSM binding to the gp130 receptor and inhibit OSM-mediated signaling.
- Chemical modification and truncation of the aptamer to enhance stability against nucleases.
Main Results:
- A high-affinity RNA aptamer, ADR58, was identified with a 7 nM affinity for human OSM.
- ADR58 effectively antagonized OSM binding to its receptor (gp130) and inhibited OSM-mediated signaling.
- A truncated and chemically modified version of ADR58 (33 bases, 2' fluorine, 2' O-methyl substitutions) retained full affinity and functional activity.
Conclusions:
- The modified ADR58 aptamer is a potent inhibitor of OSM activity.
- This aptamer serves as a valuable tool for studying OSM's role in inflammation.
- The aptamer demonstrates potential as a therapeutic agent for inflammatory conditions like rheumatoid arthritis.