CBL-b E3 ligase-mediated neddylation and activation of PARP-1 induce vascular calcification

Abstract

Vascular calcification (VC) refers to the accumulation of mineral deposits on the walls of arteries and veins, and it is closely associated with increased mortality in cardiovascular disease patients, particularly among high-risk patients with diabetes and chronic kidney disease (CKD). Neuronal precursor cell-expressed developmentally downregulated protein 8 (NEDD8) is a ubiquitin-like protein that plays a pivotal role in various cellular functions, primarily through its conjugation to target proteins and subsequent relay of biological signals. However, the role of NEDDylation in VC has not been investigated. In our study, we observed that MLN4924, an inhibitor of the NEDD8-activating E1 enzyme, effectively impedes the progression of VC. LC‒MS/MS analysis revealed that poly(ADP‒ribose) polymerase 1 (PARP-1) is subjected to NEDD8 conjugation, leading to an increase in PARP-1 activity during VC. We subsequently revealed that PARP-1 NEDDylation is mediated by the E3 ligase CBL proto-oncogene B (CBL-b) and is reversed by NEDD8-specific protease 1 (NEDP-1) during VC. Furthermore, the CBL-b C373 peptide effectively mitigated the inactive form of the E3 ligase activity of CBL-b, ultimately preventing VC. These findings provide compelling evidence that the NEDD8-dependent activation of PARP-1 represents a novel mechanism underlying vascular calcification and suggests a promising new therapeutic target for VC. Vascular calcification, a condition where calcium deposits in blood vessels, can increase heart disease risk. Researchers studied a process called neddylation, which modifies proteins, to see if it affects this calcium buildup. They experimented on cells and mice using various techniques like chemical treatments and gene alteration. They found that a protein, NEDD8, when joined to another protein, PARP-1, encourages calcium deposition in blood vessels. By preventing NEDD8 from joining to PARP-1 with a specific inhibitor, they reduced calcium buildup in cells and mice. They also found that an enzyme, Cbl-b, helps NEDD8 attach to PARP-1, suggesting a new way to prevent vascular calcification. Results suggest that stopping neddylation, particularly the joining of NEDD8 to PARP-1, can significantly reduce vascular calcification. This discovery could lead to new treatments for vascular calcification and related heart conditions. This summary was initially drafted using artificial intelligence, then revised and fact-checked by the author.