P2X7R deficiency alleviates cardiac senescence by enhancing mitophagy via the HuR/TRIM26/NR4A1 axis

Abstract

Background: Ageing is a significant risk factor for pathophysiological alterations in the heart, but the intrinsic mechanisms by which these occur have yet to be fully elucidated. Purinergic 2×7 receptor (P2X7R) is important for the pathogenesis of numerous cardiovascular diseases; nevertheless, its function in the process of cardiac ageing remains uncertain.

Methods: This study utilised P2X7R knockout (P2X7R⁻/⁻) mice. An ageing model was established by either maintaining mice until they reached 20 months of age or performing chronic subcutaneous injection of D-galactose (D-gal). Recombinant adeno-associated virus serotype 9 (AAV9) was employed to achieve cardiac-specific overexpression of P2X7R and nuclear receptor subfamily 4 group A member 1 (NR4A1). Cardiac function and histopathological changes in cardiac tissues were evaluated. Transcriptome sequencing was further applied to elucidate the potential mechanisms of P2X7R in cardiac senescence.

Result: Our result show that serum levels of P2X7R increase with advancing age in humans and that P2X7R expression is upregulated during cardiac senescence in mice. P2X7R deficiency alleviates ageing-related cardiac dysfunction, senescence phenotypes and impaired mitophagy. Cardiomyocyte-specific overexpression of P2X7R with AAV9 exacerbates the myocardial dysfunction, senescence phenotype and mitophagy disruption induced by D-gal. Mechanistically, P2X7R promotes human antigen R (HuR) nucleocytoplasmic shuttling in ageing hearts, thereby increasing the mRNA stability of tripartite motif containing 26 (TRIM26) and the expression of the E3 ubiquitin ligase TRIM26. TRIM26 subsequently mediates NR4A1 ubiquitination, leading to its proteasomal degradation, which subsequently suppresses mitophagy in cardiomyocytes and ultimately accelerates cardiac ageing.

Conclusions: Our findings provide valuable insights into the role of P2X7R in cardiac ageing and identify the HuR/TRIM26/NR4A1 axis as a key signalling pathway through which P2X7R regulates cardiac ageing.

Keywords

ageing / cardiac remodelling / human antigen R / mitophagy / NR4A1 / P2X7R / TRIM26

References

[1] Ren J, Song M, Zhang W, et al. The aging biomarker consortium represents a new era for aging research in China. Nat Med. 2023; 29: 2162-2165.

[2] Vasan RS, Enserro DM, Beiser AS, Xanthakis V. Lifetime risk of heart failure among participants in the Framingham study. J Am Coll Cardiol. 2022; 79: 250-263.

[3] Redfield MM. Heart failure with preserved ejection fraction. N Engl J Med. 2016; 375: 1868-1877.

[4] Anderson R, Lagnado A, Maggiorani D, et al. Length-independent telomere damage drives post-mitotic cardiomyocyte senescence. EMBO J. 2019; 38:e100492.

[5] Baker DJ, Childs BG, Durik M, et al. Naturally occurring p16(Ink4a)-positive cells shorten healthy lifespan. Nature. 2016; 530: 184-189.

[6] Li J, Li SH, Dong J, et al. Long-term repopulation of aged bone marrow stem cells using young Sca-1 cells promotes aged heart rejuvenation. Aging Cell. 2019; 18:e13026.

[7] Guo Y, Guan T, Shafiq K, et al. Mitochondrial dysfunction in aging. Ageing Res Rev. 2023; 88:101955.

[8] Macedo APA, da Silva ASR, Munoz VR, Ropelle ER, Pauli JR. Mitochondrial dysfunction plays an essential role in remodeling aging adipose tissue. Mech Ageing Dev. 2021; 200:111598.

[9] Lu Y, Li Z, Zhang S, Zhang T, Liu Y, Zhang L. Cellular mitophagy: mechanism, roles in diseases and small molecule pharmacological regulation. Theranostics. 2023; 13: 736-766.

[10] Hong W, Zeng X, Ma R, et al. Age-associated reduction in ER-mitochondrial contacts impairs mitochondrial lipid metabolism and autophagosome formation in the heart. Cell Death Differ. 2025; 32: 1900-1914.

[11] Wilkaniec A, Cieslik M, Murawska E, et al. P2×7 receptor is involved in mitochondrial dysfunction induced by extracellular alpha synuclein in neuroblastoma SH-SY5Y cells. Int J Mol Sci. 2020; 21: 3959.

[12] Hu SQ, Hu JL, Zou FL, et al. P2×7 receptor in inflammation and pain. Brain Res Bull. 2022; 187: 199-209.

[13] Kong H, Zhao H, Chen T, Song Y, Cui Y. Targeted P2×7/NLRP3 signaling pathway against inflammation, apoptosis, and pyroptosis of retinal endothelial cells in diabetic retinopathy. Cell Death Dis. 2022; 13: 336.

[14] Zhong X, Wang K, Wang Y, et al. Angiotension II directly bind P2×7 receptor to induce myocardial ferroptosis and remodeling by activating human antigen R. Redox Biol. 2024; 72:103154.

[15] Wang T, Li J, Li H, et al. Aerobic exercise inhibited P2×7 purinergic receptors to improve cardiac remodeling in mice with type 2 diabetes. Front Physiol. 2022; 13:828020.

[16] Ye T, Zhou Y, Yang J, et al. P2×7 receptor inhibition prevents atrial fibrillation in rodent models of depression. Europace. 2024; 26:euae022.

[17] Zhang T, Ma R, Li Z, et al. Nur77 alleviates cardiac fibrosis by upregulating GSK-3β transcription during aging. Eur J Pharmacol. 2024; 965:176290.

[18] Yao Y, Wang B, Yu K, et al. Nur77 improves ovarian function in reproductive aging mice by activating mitophagy and inhibiting apoptosis. Reprod Biol Endocrinol. 2024; 22: 86.

[19] Picca A, Faitg J, Auwerx J, Ferrucci L, D'Amico D. Mitophagy in human health, ageing and disease. Nat Metab. 2023; 5: 2047-2061.

[20] Birch J, Gil J. Senescence and the SASP: many therapeutic avenues. Genes Dev. 2020; 34: 1565-1576.

[21] Zeng L, Zhang X, Huang Z, et al. Ubiquitin proteasome system in cardiac fibrosis. J Adv Res. 2025; 76: 501-510.

[22] Hu M, Luo Q, Alitongbieke G, et al. Celastrol-induced Nur77 interaction with TRAF2 alleviates inflammation by promoting mitochondrial ubiquitination and autophagy. Mol Cell. 2017; 66: 141-153.e6.

[23] Tamiato A, Tombor LS, Fischer A, et al. Age-dependent RGS5 loss in pericytes induces cardiac dysfunction and fibrosis. Circ Res. 2024; 134: 1240-1255.

[24] Aging Biomarker Consortium, Bao H, Cao J, et al. Biomarkers of aging. Sci China Life Sci. 2023; 66: 893-1066.

[25] He Z, Moreno JA, Swain M, Wu J, Kwon O. Aminodealkenylation: ozonolysis and copper catalysis convert C(sp³)‒C(sp²) bonds to C(sp³)‒N bonds. Science. 2023; 381: 877-886.

[26] Gao H, Wang L, Lyu Y, et al. The P2X7R/NLRP3 inflammasome axis suppresses enthesis regeneration through inflammatory and metabolic macrophage-stem cell cross-talk. Sci Adv. 2025; 11:eadr4894.

[27] Huang S, Wang W, Li L, et al. P2×7 receptor deficiency ameliorates STZ-induced cardiac damage and remodeling through PKCβ and ERK. Front Cell Dev Biol. 2021; 9:692028.

[28] Yin S, Shen M, Zhang Y, et al. Nur77 increases mitophagy and decreases aggregation of α-synuclein by modulating the p-c-Abl/p-PHB2 Y121 in α-synuclein PFF SH-SY5Y cells and mice. Eur J Med Chem. 2024; 268:116251.

[29] Peng SZ, Chen XH, Chen SJ, et al. Phase separation of Nur77 mediates celastrol-induced mitophagy by promoting the liquidity of p62/SQSTM1 condensates. Nat Commun. 2021; 12: 5989.

[30] Hu M, Alitongbieke G, Su Y, Zhou H, Zhang XK. Moving nuclear receptor Nur77 to damaged mitochondria for clearance by mitophagy. Mol Cell Oncol. 2018; 5:e1327005.

[31] Hirsch TI, Wang SZ, Fligor SC, et al. Fat malabsorption in short bowel syndrome: a review of pathophysiology and management. Nutr Clin Pract. 2024; 39(suppl 1): S17-S28.

[32] Hu H, Li R, He K, et al. Bruceine A protects nuclear receptor 4A1 from ubiquitin-degradation to alleviate mesangial proliferative glomerulonephritis. Signal Transduct Target Ther. 2025; 10: 397.

[33] Cai X, Jiang Y, Cao Z, et al. Mst1-mediated phosphorylation of Nur77 improves the endometrial receptivity in human and mice. EBioMedicine. 2023; 88:104433.

[34] Di Rienzo M, Romagnoli A, Antonioli M, Piacentini M, Fimia GM. TRIM proteins in autophagy: selective sensors in cell damage and innate immune responses. Cell Death Differ. 2020; 27: 887-902.

[35] Zhu Y, Afolabi LO, Wan X, Shim JS, Chen L. TRIM family proteins: roles in proteostasis and neurodegenerative diseases. Open Biol. 2022; 12:220098.

[36] Zhang JR, Li XX, Hu WN, Li CY. Emerging role of TRIM family proteins in cardiovascular disease. Cardiology. 2020; 145: 390-400.

[37] Yang PB, Hou PP, Liu FY, et al. Blocking PPARγ interaction facilitates Nur77 interdiction of fatty acid uptake and suppresses breast cancer progression. Proc Natl Acad Sci U S A. 2020; 117: 27412-27422.

[38] Mahlokozera T, Patel B, Chen H, et al. Competitive binding of E3 ligases TRIM26 and WWP2 controls SOX2 in glioblastoma. Nat Commun. 2021; 12: 6321.

[39] Finan JM, Sutton TL, Dixon DA, Brody JR. Targeting the RNA-binding protein HuR in cancer. Cancer Res. 2023; 83: 3507-3516.

[40] Majumder M, Chakraborty P, Mohan S, Mehrotra S, Palanisamy V. HuR as a molecular target for cancer therapeutics and immune-related disorders. Adv Drug Deliv Rev. 2022; 188:114442.

[41] Ma Q, Lu Q, Lei X, et al. Relationship between HuR and tumor drug resistance. Clin Transl Oncol. 2023; 25: 1999-2014.

[42] Lal S, Burkhart RA, Beeharry N, et al. HuR posttranscriptionally regulates WEE1: implications for the DNA damage response in pancreatic cancer cells. Cancer Res. 2014; 74: 1128-1140.

[43] Guo T, Dai Z, You K, et al. S-adenosylmethionine upregulates the angiotensin receptor-binding protein ATRAP via the methylation of HuR in NAFLD. Cell Death Dis. 2021; 12: 306.

[44] Meng L, Wu H, Wu J, et al. Aberrant CircTMEM45A facilitates inflammatory progression of esophageal squamous cell carcinoma through m5C-mediated NLRP3 activation. Cancer Res. 2025; 85: 2694-2713.

[45] Adu-Amankwaah J, Bushi A, Tan R, et al. Estradiol mitigates stress-induced cardiac injury and inflammation by downregulating ADAM17 via the GPER-1/PI3K signaling pathway. Cell Mol Life Sci. 2023; 80: 246.

[46] Cittadini A, Isidori AM, Salzano A. Testosterone therapy and cardiovascular diseases. Cardiovasc Res. 2022; 118: 2039-2057.

[47] Huang JC, Duan CC, Jin S, et al. HB-EGF induces mitochondrial dysfunction via estrogen hypersecretion in granulosa cells dependent on cAMP-PKA-JNK/ERK-Ca(2+)-FOXO1 pathway. Int J Biol Sci. 2022; 18: 2047-2059.

[48] Biason-Lauber A, Lang-Muritano M. Estrogens: two nuclear receptors, multiple possibilities. Mol Cell Endocrinol. 2022; 554:111710.

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2026 The Author(s). Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.