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image of Proteome-Wide Acetylome Profiling Suggests Extensive Aspirin-Driven Remodeling of Networks Relevant to THP-1 Macrophage Differentiation

Abstract

Introduction

Acetylsalicylic Acid (aspirin or ASA) is known to exhibit immunomodulatory effects not only through Cyclooxygenase (COX) inhibition but also through direct protein acetylation. However, its impact on innate immune cell differentiation remains unclear.

Materials and Methods

To fill this gap, the study used quantitative acetyl-proteomics to track changes in the lysine acetylome during THP-1 monocyte differentiation into macrophages following ASA preconditioning.

Results

Our results showed that preconditioning of THP-1 macrophages with 300 μg/ml ASA for 3 hours before differentiation induced persistent acetylation changes. We identified 5,199 differentially acetylated sites across 2,678 proteins, with 2,595 sites upregulated and 2,604 downregulated. The sequence analysis revealed a strong preference of ASA for acidic residues like E_K motifs and hydrophobic regions. The subcellular localization analysis showed notable enrichment in the nucleus (1,166 proteins), cytoplasm (850 proteins), and mitochondria (405 proteins), and frequently contained functional domains like PWWP, SET, RhoGEF, and RNA recognition motifs.

Discussion

The Gene Ontology analysis linked these proteins to cellular metabolism, regulation, and stimulus response, while our KEGG analysis connected them to neurodegeneration, infection, and metabolic pathways. Furthermore, the protein-protein interaction networks further showed coordinated changes in ribosomal, signaling, and chromatin complexes.

Conclusion

The findings show that ASA preconditioning leaves a lasting acetylome signature during macrophage differentiation reprogramming regulatory networks relevant to macrophage differentiation and functional networks motif-directed acetylation. The results provide a plausible COX-independent model in which structural motif-targeted acetylation may underlie ASA’s immunomodulatory role.

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2026-02-25
2026-03-02

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References

  1. Kovacevic N. Beciragic D. Causevic M. Acetylsalicylic acid (aspirin): Past, present, and future. Sarajevo Med. J. 1 2 92 104 10.70119/0015‑24
    [Google Scholar]
  2. Alegbeleye B.J. Akpoveso O.O.P. Mohammed R.K. Asare B.Y.A. Pharmacology, pharmaceutics and clinical use of aspirin: A narrative review. J. Drug Deliv. Ther. 2020 10 5-s 236 253 10.22270/jddt.v10i5‑s.4351
    [Google Scholar]
  3. Rainsford K.D. Aspirin and related drugs. CRC Press 2004 10.1201/9780203646960
    [Google Scholar]
  4. Ahn T.G. Hwang J.Y. Preeclampsia and aspirin. Obstet. Gynecol. Sci. 2023 66 3 120 132 10.5468/ogs.22261 36924072
    [Google Scholar]
  5. Ishola A.A. Perspective Chapter: Aspirin - The Wonder Drug. Pain Management - From Acute to Chronic and Beyond IntechOpen 2024 10.5772/intechopen.111571
    [Google Scholar]
  6. Lucido M.J. Orlando B.J. Vecchio A.J. Malkowski M.G. Crystal structure of aspirin-acetylated human cyclooxygenase-2: Insight into the formation of products with reversed stereochemistry. Biochemistry 2016 55 8 1226 1238 10.1021/acs.biochem.5b01378 26859324
    [Google Scholar]
  7. Smith J.B. Willis A.L. Aspirin selectively inhibits prostaglandin production in human platelets. Nat. New Biol. 1971 231 25 235 237 10.1038/newbio231235a0 5284361
    [Google Scholar]
  8. Patrono C. Baigent C. Hirsh J. Roth G. Antiplatelet drugs: American college of chest physicians evidence-based clinical practice guidelines (8th Edition). Chest 2008 133 6 199S 233S (Suppl.) 10.1378/chest.08‑0672 18574266
    [Google Scholar]
  9. Fromm K. Ortelli M. Boegli A. Dehio C. Translocation of YopJ family effector proteins through the VirB/VirD4 T4SS of Bartonella. Proc. Natl. Acad. Sci. USA 2024 121 20 2310348121 10.1073/pnas.2310348121 38709922
    [Google Scholar]
  10. Xie L. Li C. Wang C. Wu Z. Wang C. Chen C. Chen X. Zhou D. Zhou Q. Lu P. Ding C. Liu C.Y. Lin J. Zhang X. Yu X. Yu W. Aspirin‐mediated acetylation of SIRT1 maintains intestinal immune homeostasis. Adv. Sci. 2024 11 19 2306378 10.1002/advs.202306378 38482749
    [Google Scholar]
  11. Wang J. Zhang C.J. Zhang J. He Y. Lee Y.M. Chen S. Lim T.K. Ng S. Shen H.M. Lin Q. Mapping sites of aspirin-induced acetylations in live cells by quantitative acid-cleavable activity-based protein profiling (QA-ABPP). Sci. Rep. 2015 5 1 7896 10.1038/srep07896 25600173
    [Google Scholar]
  12. Marimuthu S. Chivukula R.S. Alfonso L.F. Moridani M. Hagen F.K. Bhat G.J. Aspirin acetylates multiple cellular proteins in HCT-116 colon cancer cells: Identification of novel targets. Int. J. Oncol. 2011 39 5 1273 1283 10.3892/ijo.2011.1113 21743961
    [Google Scholar]
  13. Kopp E. Ghosh S. Inhibition of NF-κ B by sodium salicylate and aspirin. Science 1994 265 5174 956 959 10.1126/science.8052854 8052854
    [Google Scholar]
  14. Zhang X. Du R. Luo N. Xiang R. Shen W. Aspirin mediates histone methylation that inhibits inflammation-related stemness gene expression to diminish cancer stemness via COX-independent manner. Stem Cell Res. Ther. 2020 11 1 370 10.1186/s13287‑020‑01884‑4 32854760
    [Google Scholar]
  15. Rasheed A. Rayner K.J. Macrophage responses to environmental stimuli during homeostasis and disease. Endocr. Rev. 2021 42 4 407 435 10.1210/endrev/bnab004 33523133
    [Google Scholar]
  16. Weidenbusch M. Anders H.J. Tissue microenvironments define and get reinforced by macrophage phenotypes in homeostasis or during inflammation, repair and fibrosis. J. Innate Immun. 2012 4 5-6 463 477 10.1159/000336717 22507825
    [Google Scholar]
  17. Shah P.T. Tufail M. Wu C. Xing L. THP-1 cell line model for tuberculosis: A platform for in vitro macrophage manipulation. Tuberculosis 2022 136 102243 10.1016/j.tube.2022.102243 35963145
    [Google Scholar]
  18. Jo H. Lee E.Y. Cho H.S. Rayhan M.A. Cho A. Chae C.S. You H.J. THP-1 monocytic cells are polarized to more antitumorigenic macrophages by serial treatment with phorbol-12-myristate-13-acetate and PD98059. Medicina 2024 60 6 1009 10.3390/medicina60061009 38929626
    [Google Scholar]
  19. Ghiboub M. Zhao J. Li Yim A.Y.F. Schilderink R. Verseijden C. van Hamersveld P.H.P. Duarte J.M. Hakvoort T.B.M. Admiraal I. Harker N.R. Tough D.F. Henneman P. de Winther M.P.J. de Jonge W.J. HDAC3 mediates the inflammatory response and LPS tolerance in human monocytes and macrophages. Front. Immunol. 2020 11 550769 10.3389/fimmu.2020.550769 33123128
    [Google Scholar]
  20. Dong Z. Li R. Xu L. Xin K. Xu Y. Shi H. Sun A. Ge J. Histone hyperacetylation mediates enhanced IL‐1β production in LPS/IFN‐γ‐stimulated macrophages. Immunology 2020 160 2 183 197 10.1111/imm.13183 32061096
    [Google Scholar]
  21. Das Gupta K. Shakespear M.R. Iyer A. Fairlie D.P. Sweet M.J. Histone deacetylases in monocyte/macrophage development, activation and metabolism: Refining HDAC targets for inflammatory and infectious diseases. Clin. Transl. Immunology 2016 5 1 62 10.1038/cti.2015.46 26900475
    [Google Scholar]
  22. van den Bosch T. Boichenko A. Leus N.G.J. Ourailidou M.E. Wapenaar H. Rotili D. Mai A. Imhof A. Bischoff R. Haisma H.J. Dekker F.J. The histone acetyltransferase p300 inhibitor C646 reduces pro-inflammatory gene expression and inhibits histone deacetylases. Biochem. Pharmacol. 2016 102 130 140 10.1016/j.bcp.2015.12.010 26718586
    [Google Scholar]
  23. Bordeaux J. Welsh A.W. Agarwal S. Killiam E. Baquero M.T. Hanna J.A. Anagnostou V.K. Rimm D.L. Antibody validation. Biotechniques 2010 48 3 197 209 10.2144/000113382 20359301
    [Google Scholar]
  24. Hogrebe A. von Stechow L. Bekker-Jensen D.B. Weinert B.T. Kelstrup C.D. Olsen J.V. Benchmarking common quantification strategies for large-scale phosphoproteomics. Nat. Commun. 2018 9 1 1045 10.1038/s41467‑018‑03309‑6 29535314
    [Google Scholar]
  25. Zhang Z. Pan H. Chen X. Mass spectrometry for structural characterization of therapeutic antibodies. Mass Spectrom. Rev. 2009 28 1 147 176 10.1002/mas.20190 18720354
    [Google Scholar]
  26. Zhao M. Jia S. Gao X. Qiu H. Wu R. Wu H. Lu Q. Comparative analysis of global proteome and lysine acetylome between Naive CD4+ T cells and CD4+ T follicular helper cells. Front. Immunol. 2021 12 643441 10.3389/fimmu.2021.643441 33841426
    [Google Scholar]
  27. Zhu C. Duan Y. Dong J. Jia H. Zhang L. Xing A. Li Z. Du B. Sun Q. Huang Y. Zhang Z. Pan L. Quantitative analysis of the lysine acetylome reveals the role of SIRT3-mediated HSP60 deacetylation in suppressing intracellular Mycobacterium tuberculosis survival. Microbiol. Spectr. 2024 12 8 e00749 e24 10.1128/spectrum.00749‑24 38916288
    [Google Scholar]
  28. Alfonso L. Ai G. Spitale R.C. Bhat G.J. Molecular targets of aspirin and cancer prevention. Br. J. Cancer 2014 111 1 61 67 10.1038/bjc.2014.271 24874482
    [Google Scholar]
  29. Wang Y. Abrol R. Mak J.Y.W. Das Gupta K. Ramnath D. Karunakaran D. Fairlie D.P. Sweet M.J. Histone deacetylase 7: A signalling hub controlling development, inflammation, metabolism and disease. FEBS J. 2023 290 11 2805 2832 10.1111/febs.16437 35303381
    [Google Scholar]
  30. Kornak U. Bischof O. Hesse E. Jakob F. Ebert R. Taipaleenmäki H. Epigenetics and noncoding RNA - Principles and clinical impact. Osteologie 2021 30 03 201 210 10.1055/a‑1527‑4585
    [Google Scholar]
  31. Franklin K.A. Shields C.E. Haynes K.A. Beyond the marks: Reader-effectors as drivers of epigenetics and chromatin engineering. Trends Biochem. Sci. 2022 47 5 417 432 10.1016/j.tibs.2022.03.002 35427480
    [Google Scholar]
/content/journals/ppl/10.2174/0109298665444488260123052552
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Proteome-Wide Acetylome Profiling Suggests Extensive Aspirin-Driven Remodeling of Networks Relevant to THP-1 Macrophage Differentiation
Bentham Science Publishers ; https://doi.org/10.2174/0109298665444488260123052552
/content/journals/ppl/10.2174/0109298665444488260123052552
/content/journals/ppl/10.2174/0109298665444488260123052552
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  • Article Type:     Research Article
Keywords: Aspirin (acetylsalicylic acid) ; immunomodulation ; post-translational modification ; THP-1 cells ; acetylome profiling ; lysine acetylation

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