An aptamer agonist of the insulin receptor acts as a positive or negative allosteric modulator, depending on its concentration
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Aptamers are widely used as binders that interact with targets with high affinity or as inhibitors of the function of target molecules. However, they have also been used to modulate target protein function, which they achieve by activating the target or stabilizing its conformation. Here, we report a unique aptamer modulator of the insulin receptor (IR), IR-A62. Alone, IR-A62 acts as a biased agonist that preferentially induces Y1150 monophosphorylation of IR. However, when administered alongside insulin, IR-A62 shows variable binding cooperativity depending on the ligand concentration. At low concentrations, IR-A62 acts as a positive allosteric modulator (PAM) agonist that enhances insulin binding, but at high concentrations, it acts as a negative allosteric modulator (NAM) agonist that competes with insulin for IR. Moreover, the concentration of insulin affects the binding of IR-A62 to IR. Finally, the subcutaneous administration of IR-A62 to diabetic mice reduces blood glucose levels with a longer-lasting effect than insulin administration. These findings imply that aptamers can elicit various responses from receptors beyond those of a simple agonist or inhibitor. We expect further studies of IR-A62 to help reveal the mechanism of IR activation and greatly expand the range of therapeutic applications of aptamers.
Abstract
Aptamers are widely used as binders that interact with targets with high affinity or as inhibitors of the function of target molecules. However, they have also been used to modulate target protein function, which they achieve by activating the target or stabilizing its conformation. Here, we report a unique aptamer modulator of the insulin receptor (IR), IR-A62. Alone, IR-A62 acts as a biased agonist that preferentially induces Y1150 monophosphorylation of IR. However, when administered alongside insulin, IR-A62 shows variable binding cooperativity depending on the ligand concentration. At low concentrations, IR-A62 acts as a positive allosteric modulator (PAM) agonist that enhances insulin binding, but at high concentrations, it acts as a negative allosteric modulator (NAM) agonist that competes with insulin for IR. Moreover, the concentration of insulin affects the binding of IR-A62 to IR. Finally, the subcutaneous administration of IR-A62 to diabetic mice reduces blood glucose levels with a longer-lasting effect than insulin administration. These findings imply that aptamers can elicit various responses from receptors beyond those of a simple agonist or inhibitor. We expect further studies of IR-A62 to help reveal the mechanism of IR activation and greatly expand the range of therapeutic applications of aptamers.
참고문헌 (29)
- Zhou, J., "Aptamers as targeted therapeutics : current potential and challenges", Nat. Rev. Drug Discov, 16, pp.181~202 (2017)
- Ellington, A. D., "In vitro selection of RNA molecules that bind specific ligands", Nature, 346, pp.818~822 (1990)
- Tuerk, C., "Systematic evolution of ligands by exponential enrichment : RNA ligands to bacteriophage T4 DNA polymerase", Science, 249, pp.505~510 (1990)
- Gelinas, A. D., "Embracing proteins: structural themes in aptamer-protein complexes", Curr. Opin. Struct. Biol, 36, pp.122~132 (2016)
- Nimjee, S. M., "Aptamers as therapeutics", Annu. Rev. Pharmacol. Toxicol, 57, pp.61~79 (2017)
- Dollins, C. M., "Assembling OX40 aptamers on a molecular scaffold to create a receptor-activating aptamer", Chem. Biol, 15, pp.675~682 (2008)
- McNamara, J. O., "Multivalent 4-1BB binding aptamers costimulate CD8+T cells and inhibit tumor growth in mice", J. Clin. Invest, 118, pp.376~386 (2008)
- Ramaswamy, V., "DNA aptamer assembly as a vascular endothelial growth factor receptor agonist", Nucleic Acid Ther, 25, pp.227~234 (2015)
- Ueki, R., "A chemically unmodified agonistic DNA with growth factor functionality for in vivo therapeutic application", Sci. Adv, 6, pp.eaay2801~ (2020)
- Huang, Y. Z., "RNA aptamer-based functional ligands of the neurotrophin receptor, TrkB", Mol. Pharmacol, 82, pp.623~635 (2012)
- Yunn, N. -O., "Agonistic aptamer to the insulin receptor leads to biased signaling and functional selectivity through allosteric modulation", Nucleic Acids Res, 43, pp.7688~7701 (2015)
- Kahsai, A. W., "Conformationally selective RNA aptamers allosterically modulate the β2-adrenoceptor", Nat. Chem. Biol, 12, pp.709~716 (2016)
- Takahashi, M., "Nucleic acid ligands act as a PAM and agonist depending on the intrinsic ligand binding state of P2RY2", Proc. Natl Acad. Sci. USA, 118, pp.e2019497118~ (2021)
- Yunn, N. -O., "A hotspot for enhancing insulin receptor activation revealed by a conformation-specific allosteric aptamer", Nucleic Acids Res, 49, pp.700~712 (2021)
- Belfiore, A., "Insulin receptor isoforms in physiology and disease : an updated view", Endocr. Rev, 38, pp.379~431 (2017)
- Lollo, B., "Beyond antibodies : new affinity reagents to unlock the proteome", Proteomics, 14, pp.638~644 (2014)
- Gao, S., "Post-SELEX optimization of aptamers", Anal. Bioanal. Chem, 408, pp.4567~4573 (2016)
- Gupta, S., "Pharmacokinetic properties of DNA aptamers with base modifications", Nucleic Acid Ther, 27, pp.345~353 (2017)
- Haeusler, R. A., "Biochemical and cellular properties of insulin receptor signalling", Nat. Rev. Mol. Cell Biol, 19, pp.31~44 (2017)
- Huang, X., "The PI3K", AKT pathway in obesity and type 2diabetes, 2018, pp.14, 1483~1496
- Belfiore, A., "Insulin receptor and cancer", Endocr. Relat. Cancer, 18, pp.R125~R147 (2011)
- Sah, S. P., "Animal models of insulin resistance:a review", Pharmacol. Rep, 68, pp.1165~1177 (2016)
- Hubbard, S. R., "Protein tyrosine kinase structure and function", Annu. Rev. Biochem, 69, pp.373~398 (2000)
- McKern, N. M., "Structure of the insulin receptor ectodomain reveals a foldedover conformation", Nature, 443, pp.218~221 (2006)
- Croll, T. I., "Higher-resolution structure of the human insulin receptor ectodomain: multi-modal inclusion of the insert domain", Structure, 24, pp.469~476 (2016)
- Weis, F., "The signalling conformation of the insulin receptor ectodomain", Nat. Commun, 9, pp.4420~ (2018)
- Uchikawa, E., "Activation mechanism of the insulin receptor revealed by cryo-EM structure of the fully liganded receptorligand complex", eLife, 8, pp.e48630~ (2019)
- Giesbertz, P., "Metabolite profiling in plasma and tissues of ob", ob and db, Diabetologia, pp.2015, 58[DOI]
- Zhang, Y., "Recent advances in aptamer discovery and applications", Molecules, 24, pp.941~ (2019)