The generation and characterisation of antagonist RNA aptamers to MCP-1.
Monocyte chemoattractant protein-1 (MCP-1) has been implicated as a powerful pro-inflammatory mediator and may represent a potentially important, therapeutic opportunity for treatment of inflammatory disease and atherosclerosis. To further investigate the role of MCP-1 in inflammatory disorders we have isolated a series of RNA aptamers that bind specifically to mouse MCP-1. The highest affinity aptamers, designated ADR7 and ADR22, have been functionally characterised in vitro and in cell based assays. ADR7 and ADR22 have an affinity of 180 pM and 370 pM respectively for mouse MCP-1, they can antagonise MCP-1 binding to heparin and specifically antagonise MCP-1 induced chemotaxis in a cell based assay. An interesting feature of ADR22 but not ADR7 is that it is capable of antagonising the function of human MCP-1, demonstrating the high level of specificity of these aptamers and that the aptamers recognise MCP-1 in different ways. The aptamers may be used as a tool to further investigate the role of MCP-1 in inflammatory disorders and may also have a role as a therapeutic agent.
Rhodes
A
A
Molecular Discovery Department, Glaxo Wellcome Research and Development, Stevenage, Herts SG1 2NY, UK. adr7003@gsk.com
Smithers
N
N
Chapman
T
T
Parsons
S
S
Rees
S
S
D000818:Animals; D000925:Anticoagulants / Q000378:metabolism; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism; D018932:Chemokine CCL2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002633:Chemotaxis / Q000502:physiology; D006224:Cricetinae; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008024:Ligands; D051379:Mice; D011485:Protein Binding; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D054390:Receptors, CCR2; D019707:Receptors, Chemokine / Q000378:metabolism; D020349:Surface Plasmon Resonance
D000925:Anticoagulants; C515334:CCR2 protein, human; C515335:Ccr2 protein, mouse; D018932:Chemokine CCL2; D008024:Ligands; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D012313:RNA; D006493:Heparin; D002118:Calcium