Identification of potent and selective RNA antagonists of the IFN-gamma-inducible CXCL10 chemokine.
CXCL10 (also known as IP-10 in humans and CRG-2 in mice) is a nonglycosylated chemokine and a member of the non-ELR CXC chemokine subfamily implicated in a variety of inflammatory conditions. The role of CXCL10 in different disease states still requires clarification, and new approaches are necessary to better understand its biological function. We report here the isolation of a series of nuclease-resistant RNA aptamers that act to antagonize human CXCL10 function in a number of in vitro and cell-based assays. The two most potent aptamers identified were highly selective for human CXCL10. A further aptamer was identified that antagonized both the human and the mouse CXCL10. A combination of a molecular-biology-based truncation and solid-phase synthesis enabled the truncation of one of the aptamers from 71 to 34 nucleotides. This was followed by PEGylation, 3' capping, and further stabilization of the RNA aptamer, while its high potency was maintained. These aptamers could be utilized as powerful target validation tools and may also have therapeutic potential. To our knowledge, the CXCL10 aptamers generated are the most potent antagonists of CXCL10/CXCR3 signaling reported to date.
Marro
Martin L
ML
Gene Expression and Protein Biochemistry, GlaxoSmithKline Medicines Research Centre, Gunnels Wood Road, Stevenage, Herts SG1 2NY, United Kingdom.
Daniels
Dion A
DA
McNamee
Anne
A
Andrew
David P
DP
Chapman
Trevor D
TD
Jiang
Ming S
MS
Wu
Zining
Z
Smith
Janet L
JL
Patel
Kalpana K
KK
Gearing
Katy L
KL
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D016466:CHO Cells; D045744:Cell Line, Tumor; D002464:Cell Migration Inhibition; D054357:Chemokine CXCL10; D019743:Chemokines, CXC / Q000037:antagonists & inhibitors* / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D006224:Cricetinae; D006801:Humans; D007371:Interferon-gamma / Q000502:physiology*; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D011092:Polyethylene Glycols / Q000737:chemistry; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D051381:Rats; D054367:Receptors, CXCR3; D019707:Receptors, Chemokine / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology
C515187:CXCR3 protein, human; D054357:Chemokine CXCL10; D019743:Chemokines, CXC; C515188:Cxcr3 protein, mouse; C515189:Cxcr3 protein, rat; D008024:Ligands; D016601:RNA-Binding Proteins; D054367:Receptors, CXCR3; D019707:Receptors, Chemokine; D011092:Polyethylene Glycols; D012313:RNA; D007371:Interferon-gamma