Genotoxicity and immunogenicity of DNA-advanced glycation end products formed by methylglyoxal and lysine in presence of Cu2+

The highly reactive electrophile, methylglyoxal (MG), a break down product of carbohydrates, is a major environmental mutagen having potential genotoxic effects. Previous studies have suggested the reaction of MG with free amino groups of proteins forming advanced glycation end products (AGEs). This results in the generation of free radicals which play an important role in pathophysiology of aging and diabetic complications. MG also reacts with free amino group of nucleic acids resulting in the formation of DNA–AGEs. While the formation of nucleoside AGEs has been demonstrated previously, no extensive studies have been performed to assess the genotoxicity and immunogenicity of DNA–AGEs. In this study we report both the genotoxicity and immunogenicity of AGEs formed by MG–Lys–Cu2+ system. Genotoxicity of the experimentally generated AGEs was confirmed by comet-assay. Spectroscopical analysis and melting temperature studies suggest structural perturbations in the DNA as a result of modification. This might be due to generation of single-stranded regions and destabilization of hydrogen bonds. Immunogenicity of native and MG–Lys–Cu2+-DNA was probed in female rabbits. The modified DNA was highly immunogenic eliciting high titre immunogen specific antibodies, while the unmodified form was almost non-immunogenic. The results show structural perturbations in MG–Lys–Cu2+-DNA generating new epitopes that render the molecule immunogenic.

► MG-Lys-Cu2+ generates AGEs which were found genotoxic to human lymphocyte.
► Glycation by reactive carbonyl species generates neo-epitopes in human DNA
► Modified human DNA was found to be highly immunogenic in experimental animals.
► The induced antibodies exhibited wide range of heterogeneity and poly-specificity.
► Role of DNA-AGEs in diabetes /associated secondary complications has been discussed.