Glucosepane Crosslinks and Routes to Cleavage (Yale University)
Our arteries slowly stiffen with age. One of the key drivers of this stiffening process is the accumulation of molecular crosslinks that are formed between adjacent strands of structural proteins of the large arteries. One major cause of crosslink accumulation in aging is Advanced Glycation Endproducts (AGE), and one AGE in particular, called glucosepane, is currently thought to be the single largest contributor to tissue AGE crosslinking.
The Yale AGE team is working on new tools for the detection of AGEs and their precursors. This work has already achieved start-to-finish, high-yield synthesis of glucosepane from common laboratory reagents in only a few steps. Also being investigated are glucosepane’s peptide adducts, its precursor MGH (methylglyoxal-derived hydroimidazolone modifications of arginine), and the understudied crosslink pentosinane. The team is now using these compounds to develop new antibodies and reagents to enable rejuvenation research. One of these is small peptides containing glucosepane structures, an early step toward enabling high-throughput screening of potential glucosepane-breaking drugs.
In earlier Foundation-funded research, a group at Cambridge University found that all of the commercially-available antibodies for the major AGE-related molecules are actually highly unreliable. This is a serious impediment to detecting such molecules in tissue samples and in evaluating the effects of interventions targeting them, whether in high-throughput screening or in later studies in vivo. The Yale glucosepane team is now tackling this problem. They recently published a scientific paper describing their use of the same underlying chemistry to generate antibodies targeting the three key precursors to methylglyoxal, a toxic AGE precursor molecule implicated in the cellular dysfunction underlying diabetic neuropathy and retinopathy. These antibodies are capable of detecting the methylglyoxal precursors inside cells, enabling more powerful research on the cellular targets of these molecules and the mechanisms of their toxicity to cells.
The team is now working to do the same thing for glucosepane, using the novel chemistry they have developed. They are working to synthesize a vaccine consisting of glucosepane peptide antigens, which will be administered to rouse antibodies targeting glucosepane-containing peptides in vivo. By isolating and modifying the immune cells that generate the antibodies, the team expects to be able to produce glucosepane-targeting antibodies on an industrial scale.