Prototypic P2X7 Receptor Agonist, BzATP, Induced the Expression of Unfolded Protein Response Genes in Human M1 Macrophages
-
Maryam Akhtari
,
Seyed Jalal Zargar
,
Ali Javinani
,
Amir Ashraf-Ganjouei
,
Mahdi Vojdanian
,
Ahmadreza Jamshidi
,
Mahdi Mahmoudi
Abstract
Purinergic receptors stimulation by adenosine triphosphate (ATP) contributes significantly to macrophage activation, and also macrophage cell death. Upon the macrophage activation, the protein load of the endoplasmic reticulum is increased which is resulted in the activation of unfolded protein response (UPR). In the current study, we aimed to evaluate the connection between prototypic P2X7 receptor agonist, extracellular 2ʹ(3ʹ)-O-(4-Benzoylbenzoyl)-ATP (BzATP), and the UPR pathway in macrophages.
The monocyte-derived macrophages from blood samples of 14 healthy volunteers were skewed toward M1 macrophages after incubation with LPS and IFN-γ. M1 macrophages were treated with 200 µM BzATP. The expression levels of UPR genes, including CHOP, HERP, GADD34, XBP1, and ATF6 in macrophages before and after treatment were measured using real-time polymerase chain reaction.
The results demonstrated that the expression of CHOP, HERP, and ATF6 is significantly decreased and the expression level of GADD34 and XBP1 is significantly increased after M1 polarization. BzATP not only significantly increased the expression levels of CHOP, GADD34, ATF6, and HERP but also significantly decreases the XBP1 expression level in M1 macrophages.
The present study showed that BzATP induces cellular stress in M1 macrophages by elevating the expression levels of UPR genes including CHOP, GADD34, ATF6, and reducing cell viability.
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15. Martinon F, Chen X, Lee A-H, Glimcher LH. Toll-like receptor activation of XBP1 regulates innate immune responses in macrophages. Nat Immunol. 2010;11(5):411-9.
16. Brush MH, Weiser DC, Shenolikar S. Growth arrest and DNA damage-inducible protein GADD34 targets protein phosphatase 1α to the endoplasmic reticulum and promotes dephosphorylation of the α subunit of eukaryotic translation initiation factor 2. Mol Cell Biol. 2003;23(4):1292-303.
17. Nakayama Y, Endo M, Tsukano H, Mori M, Oike Y, Gotoh T. Molecular mechanisms of the LPS-induced non-apoptotic ER stress-CHOP pathway. J Biochem. 2010;147(4):471-83.
18. Ohtomo K, Shatos MA, Hodges RR, Dartt DA. BzATP Activates P2X7 and P2Y Receptors in Lacrimal Gland Myoepithelial Cells. Investigative Ophthalmology & Visual Science. 2010;51(13):4180-.
19. Ma Y, Hendershot LM. Herp is dually regulated by both the endoplasmic reticulum stress-specific branch of the unfolded protein response and a branch that is shared with other cellular stress pathways. J of Biol Chem. 2004;279(14):13792-9.
20. Hetz C, Papa FR. The Unfolded Protein Response and Cell Fate Control. Mol Cell. 2018;69(2):169-81.
21. Jain BP. An Overview of Unfolded Protein Response Signaling and Its Role in Cancer. Cancer Biother Radiopharm. 2017;32(8):275-81.
22. Li Y, Guo Y, Tang J, Jiang J, Chen Z. New insights into the roles of CHOP-induced apoptosis in ER stress. Acta biochim biophysi Sin. 2014;46(8):629-40.
23. Yang H, Niemeijer M, van de Water B, Beltman JB. ATF6 Is a Critical Determinant of CHOP Dynamics during the Unfolded Protein Response. iScience. 2020;23(2):100860.
24. Miao EA, Rajan JV, Aderem A. Caspase-1-induced pyroptotic cell death. Immunol Rev. 2011;243(1):206-14.
25. Le Feuvre RA, Brough D, Iwakura Y, Takeda K, Rothwell NJ. Priming of macrophages with lipopolysaccharide potentiates P2X7-mediated cell death via a caspase-1-dependent mechanism, independently of cytokine production. J Biol Chem. 2002;277(5):3210-8.
26. Hanley PJ, Kronlage M, Kirschning C, Del Rey A, Di Virgilio F, Leipziger J, et al. Transient P2X7 receptor activation triggers macrophage death independent of Toll-like receptors 2 and 4, caspase-1, and pannexin-1 proteins. J Biol Chem. 2012;287(13):10650-63.
27. Wang Q, Wang L, Feng YH, Li X, Zeng R, Gorodeski GI. P2X7 receptor-mediated apoptosis of human cervical epithelial cells. Am J Physiol Cell Physiol. 2004;287(5):C1349-58.
28. Noguchi T, Ishii K, Fukutomi H, Naguro I, Matsuzawa A, Takeda K, et al. Requirement of reactive oxygen species-dependent activation of ASK1-p38 MAPK pathway for extracellular ATP-induced apoptosis in macrophage. J Biol Chem. 2008;283(12):7657-65.
29. Lees MP, Fuller SJ, McLeod R, Boulter NR, Miller CM, Zakrzewski AM, et al. P2X7 receptor-mediated killing of an intracellular parasite, Toxoplasma gondii, by human and murine macrophages. J Immunol. 2010;184(12):7040-6.
30. Chaves SP, Torres-Santos EC, Marques C, Figliuolo VR, Persechini PM, Coutinho-Silva R, et al. Modulation of P2X(7) purinergic receptor in macrophages by Leishmania amazonensis and its role in parasite elimination. Microbes Infect. 2009;11(10-11):842-9.
31. Akhtari M, Zargar SJ, Vojdanian M, Jamshidi A, Mahmoudi M. Monocyte-derived and M1 macrophages from ankylosing spondylitis patients released higher TNF-α and expressed more IL1B in response to BzATP than macrophages from healthy subjects. Sci Rep. 2021;11(1):17842.
2. North RA. P2X receptors. Philos Trans R Soc Lond B Biol Sci. 2016;371(1700).
3. Burnstock G. P2X ion channel receptors and inflammation. Purinergic Signal. 2016;12(1):59-67.
4. Di Virgilio F, Dal Ben D, Sarti AC, Giuliani AL, Falzoni S. The P2X7 Receptor in Infection and Inflammation. Immunity. 2017;47(1):15-31.
5. Roger S, Jelassi B, Couillin I, Pelegrin P, Besson P, Jiang L-H. Understanding the roles of the P2X7 receptor in solid tumour progression and therapeutic perspectives. Biomembranes. 2015;1848(10):2584-602.
6. Lara R, Adinolfi E, Harwood CA, Philpott M, Barden JA, Di Virgilio F, et al. P2X7 in Cancer: From Molecular Mechanisms to Therapeutics. Front Pharmacol. 2020;11(793):225-9.
7. Hetz C, Zhang K, Kaufman RJ. Mechanisms, regulation and functions of the unfolded protein response. Nat Rev Mol Cell Biol. 2020;21(8):421-38.
8. Grootjans J, Kaser A, Kaufman RJ, Blumberg RS. The unfolded protein response in immunity and inflammation. Nat Rev Immunol. 2016;16(8):469-84.
9. Hetz C, Saxena S. ER stress and the unfolded protein response in neurodegeneration. Nat Rev Neurol. 2017;13(8):477-91.
10. Chao CC, Chan P, Kuo CS, Gong CL, Cheng TH, Liu ZM, et al. Protection of differentiated neuronal NG108-15 cells from P2X7 receptor-mediated toxicity by taurine. Pharmacol Rep. 2014;66(4):576-84.
11. Akhtari M, Zargar SJ, Mahmoudi M, Vojdanian M, Rezaeimanesh A, Jamshidi A. Ankylosing spondylitis monocyte-derived macrophages express increased level of A2A adenosine receptor and decreased level of ectonucleoside triphosphate diphosphohydrolase-1 (CD39), A1 and A2B adenosine receptors. Clin Rheumatol. 2018;37(6):1589-95.
12. Lari A, Gholami Pourbadie H, Jafari M, Sharifi-Zarchi A, Akhtari M, Nejatbakhsh Samimi L, et al. Downregulation of ITM2A Gene Expression in Macrophages of Patients with Ankylosing Spondylitis. Int Arch Allergy Immunol. 2021;8(4):2-6.
13. Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative CT method. Nat Protoc. 2008;3(6):1101-8.
14. Keestra-Gounder AM, Byndloss MX, Seyffert N, Young BM, Chávez-Arroyo A, Tsai AY, et al. NOD1 and NOD2 signalling links ER stress with inflammation. Nature. 2016;532(7599):394-7.
15. Martinon F, Chen X, Lee A-H, Glimcher LH. Toll-like receptor activation of XBP1 regulates innate immune responses in macrophages. Nat Immunol. 2010;11(5):411-9.
16. Brush MH, Weiser DC, Shenolikar S. Growth arrest and DNA damage-inducible protein GADD34 targets protein phosphatase 1α to the endoplasmic reticulum and promotes dephosphorylation of the α subunit of eukaryotic translation initiation factor 2. Mol Cell Biol. 2003;23(4):1292-303.
17. Nakayama Y, Endo M, Tsukano H, Mori M, Oike Y, Gotoh T. Molecular mechanisms of the LPS-induced non-apoptotic ER stress-CHOP pathway. J Biochem. 2010;147(4):471-83.
18. Ohtomo K, Shatos MA, Hodges RR, Dartt DA. BzATP Activates P2X7 and P2Y Receptors in Lacrimal Gland Myoepithelial Cells. Investigative Ophthalmology & Visual Science. 2010;51(13):4180-.
19. Ma Y, Hendershot LM. Herp is dually regulated by both the endoplasmic reticulum stress-specific branch of the unfolded protein response and a branch that is shared with other cellular stress pathways. J of Biol Chem. 2004;279(14):13792-9.
20. Hetz C, Papa FR. The Unfolded Protein Response and Cell Fate Control. Mol Cell. 2018;69(2):169-81.
21. Jain BP. An Overview of Unfolded Protein Response Signaling and Its Role in Cancer. Cancer Biother Radiopharm. 2017;32(8):275-81.
22. Li Y, Guo Y, Tang J, Jiang J, Chen Z. New insights into the roles of CHOP-induced apoptosis in ER stress. Acta biochim biophysi Sin. 2014;46(8):629-40.
23. Yang H, Niemeijer M, van de Water B, Beltman JB. ATF6 Is a Critical Determinant of CHOP Dynamics during the Unfolded Protein Response. iScience. 2020;23(2):100860.
24. Miao EA, Rajan JV, Aderem A. Caspase-1-induced pyroptotic cell death. Immunol Rev. 2011;243(1):206-14.
25. Le Feuvre RA, Brough D, Iwakura Y, Takeda K, Rothwell NJ. Priming of macrophages with lipopolysaccharide potentiates P2X7-mediated cell death via a caspase-1-dependent mechanism, independently of cytokine production. J Biol Chem. 2002;277(5):3210-8.
26. Hanley PJ, Kronlage M, Kirschning C, Del Rey A, Di Virgilio F, Leipziger J, et al. Transient P2X7 receptor activation triggers macrophage death independent of Toll-like receptors 2 and 4, caspase-1, and pannexin-1 proteins. J Biol Chem. 2012;287(13):10650-63.
27. Wang Q, Wang L, Feng YH, Li X, Zeng R, Gorodeski GI. P2X7 receptor-mediated apoptosis of human cervical epithelial cells. Am J Physiol Cell Physiol. 2004;287(5):C1349-58.
28. Noguchi T, Ishii K, Fukutomi H, Naguro I, Matsuzawa A, Takeda K, et al. Requirement of reactive oxygen species-dependent activation of ASK1-p38 MAPK pathway for extracellular ATP-induced apoptosis in macrophage. J Biol Chem. 2008;283(12):7657-65.
29. Lees MP, Fuller SJ, McLeod R, Boulter NR, Miller CM, Zakrzewski AM, et al. P2X7 receptor-mediated killing of an intracellular parasite, Toxoplasma gondii, by human and murine macrophages. J Immunol. 2010;184(12):7040-6.
30. Chaves SP, Torres-Santos EC, Marques C, Figliuolo VR, Persechini PM, Coutinho-Silva R, et al. Modulation of P2X(7) purinergic receptor in macrophages by Leishmania amazonensis and its role in parasite elimination. Microbes Infect. 2009;11(10-11):842-9.
31. Akhtari M, Zargar SJ, Vojdanian M, Jamshidi A, Mahmoudi M. Monocyte-derived and M1 macrophages from ankylosing spondylitis patients released higher TNF-α and expressed more IL1B in response to BzATP than macrophages from healthy subjects. Sci Rep. 2021;11(1):17842.
| Files | ||
| Issue | Vol 21 No 1 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijaai.v21i1.8618 | |
| Keywords | ||
| 3'-O-(4-benzoyl)benzoyladenosine 5'-triphosphate Cell death Purinergic receptors Unfolded protein response | ||
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How to Cite
1.
Akhtari M, Jalal Zargar S, Javinani A, Ashraf-Ganjouei A, Vojdanian M, Jamshidi A, Mahmoudi M. Prototypic P2X7 Receptor Agonist, BzATP, Induced the Expression of Unfolded Protein Response Genes in Human M1 Macrophages. Iran J Allergy Asthma Immunol. 2022;21(1):73-80.
