Abstract
Proteins in the eye lens have negligible turnover and therefore progressively accumulate chemical modifications during aging. Carbonyls and oxidative stresses, which are intricately linked to one another, predominantly drive such modifications. Oxidative stress leads to the loss of glutathione (GSH) and ascorbate degradation; this in turn leads to the formation of highly reactive dicarbonyl compounds that react with proteins to form advanced glycation end products (AGEs). The formation of AGEs leads to the crosslinking and aggregation of proteins contributing to lens aging and cataract formation. To inhibit AGE formation, we developed a disulfide compound linking GSH diester and mercaptoethylguanidine, and we named it carboxitin. Bovine lens organ cultured with carboxitin showed higher levels of GSH and mercaptoethylguanidine in the lens nucleus. Carboxitin inhibited erythrulose-mediated mouse lens protein crosslinking, AGE formation and the formation of 3-deoxythreosone, a major ascorbate-derived AGE precursor in the human lens. Carboxitin inhibited the glycation-mediated increase in stiffness in organ-cultured mouse lenses measured using compressive mechanical strain. Delivery of carboxitin into the lens increases GSH levels, traps dicarbonyl compounds and inhibits AGE formation. These properties of carboxitin could be exploited to develop a therapy against the formation of AGEs and the increase in stiffness that causes presbyopia in aging lenses.
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Acknowledgments
The authors thank Saving Sight, Kansas City, MO, for providing the human lenses.
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This work was supported by the National Institutes of Health Grants EY028836 and EY023286 (RHN) and an RPB challenge grant to the Department of Ophthalmology, University of Colorado.
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RHN and ML conceptualized the project. SKN, JR, SR and RBN conducted the experiments. SKN, SR, JR and RHN analyzed the results. All authors wrote the paper and approved the manuscript.
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Supplementary Fig. 1
Commercial erythrulose contains minute levels of glyoxal. (a) Reversed-phase column chromatography elusion profiles of quinoxaline, erythrulose, OPD-derivatized erythrulose and erythrulose pretreated with aminoguanidine hydrochloride (AGH) and then derivatized with OPD. The arrow points to glyoxal-Q. (b) Quantification of the glyoxal content in erythrulose. (DOCX 219 kb)
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Nandi, S.K., Rankenberg, J., Rakete, S. et al. Glycation-mediated protein crosslinking and stiffening in mouse lenses are inhibited by carboxitin in vitro. Glycoconj J 38, 347–359 (2021). https://doi.org/10.1007/s10719-020-09961-9
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DOI: https://doi.org/10.1007/s10719-020-09961-9