Aspirin Reshapes Acetylomes in Inflammatory and Cancer Cells via CoA-Dependent and CoA-Independent Pathways

Aspirin, or acetylsalicylic
acid (ASA), is the most widely used
medication to relieve pain, fever, and inflammation. Recent studies
have revealed new benefits of aspirin, including reduction of heart
attack and stroke, anticancer, and life extension. Despite the profound
effects of aspirin, the mechanism of its action remains to be elucidated.
Here, we used deuterium-labeled aspirin (D-aspirin) together with
mass spectrometry-based acetylomic analysis, termed DAcMS, to investigate
the landscape of protein acetylation induced by aspirin. The DAcMS
revealed the acetylomes of lipopolysaccharide-induced inflammatory
BV2 cells and colon cancer HCT116 cells. The acetylation level was
substantially induced upon aspirin treatment in both cell lines. In
total, we identified 17,003 acetylation sites on 4623 proteins in
BV2 cells and 16,366 acetylated sites corresponding to 4702 acetylated
proteins in HCT116 cells. Importantly, functional analyses of these
aspirin-induced acetylated proteins suggested that they were highly
enriched in many key biological categories, which function importantly
in inflammatory response. We further demonstrated that aspirin acetylates
proteins through both acetyl-CoA-dependent and acetyl-CoA-independent
pathways, and the accessible lysine residues at the protein surface
are major acetylation targets of aspirin. Hence, our study provides
the comprehensive atlas of aspirin-induced acetylome under disease
conditions. This knowledge proffers new insight into the aspirin-directed
acetylome and perhaps new drug target sites relevant to human cancer
and inflammatory diseases. The MS data of this study have been deposited
under the accession number IPX0001923000 at iProX.