Inhibition of CREB promotes glucocorticoids action on airway inflammation in pediatric asthma by promoting ferroptosis of eosinophils
Main Article Content
Keywords
airway inflammation, CREB, eosinophils, ferroptosis, glucocorticoids, pediatric asthma
Abstract
Background: Pediatric asthma is a common chronic disease of childhood with airway inflammation. Cyclic adenosine monophosphate response element binding protein (CREB) plays a significant role in the transcription of proinflammatory genes, but its role in pediatric asthma has remained unclear. Herein, we investigated the functions of CREB in pediatric asthma.
Methods: Eosinophils were purified from the peripheral blood of interleukin 5 (IL5) transgenic (IL5T) neonatal mice. The contents of CREB, long-chain fatty-acid–CoA ligase 4, transferrin receptor protein 1, ferritin heavy chain 1, and glutathione peroxidase 4 in eosinophils were examined by Western blot analysis. The viability of eosinophils, and the mean fluorescence intensity of Siglec F, C-C motif chemokine receptor 3 (CCR3), and reactive oxygen species were examined by flow cytometry. The concentration of iron in eosinophils was assessed by a commercial kit. The contents of malondialdehyde, glutathione, glutathione peroxidase, IL-5, and IL-4 were discovered by enzyme-linked-immunosorbent serologic assay. The C57BL/6 mice were randomly divided into four groups: sham, ovalbumin (OVA), OVA+Ad-shNC, and OVA+Ad-shCREB. The bronchial and alveolar structures were evaluated by hematoxylin and eosin staining. Leukocytes and eosinophils in the blood were measured using a HEMAVET 950.
Results: The abundance of CREB in eosinophils was enhanced by CREB overexpression vector transfection, but reduced by short hairpin (sh)CREB transfection. Downregulation of CREB triggered the cell death of eosinophils. Knockdown of CREB could obviously contribute to ferroptosis of eosinophils. In addition, downregulation of CREB facilitated dexamethasone (DXMS, a type of glucocorticoid)-induced eosinophils death. Moreover, we established an asthma mouse model by OVA treatment. The CREB was upregulated in OVA group mice, but Ad-shCREB treatment obviously downregulated CREB level. Downregulation of CREB diminished OVA-induced asthmatic airway inflammation by reducing the number of inflammatory cells and the levels of proinflammatory factors. Downregulated CREB enhanced the anti-inflammatory effect of DXMS in OVA-induced mice.
Conclusion: Inhibition of CREB promoted the effect of glucocorticoids on airway inflammation in pediatric asthma through promoting ferroptosis of eosinophils.
References
2. Miller RL, Grayson MH, Strothman K. Advances in asthma: New understandings of asthma’s natural history, risk factors, underlying mechanisms, and clinical management. J Allergy Clin Immunol. 2021;148(6):1430–1441. 10.1016/j.jaci.2021.10.001
3. Devonshire AL, Kumar R. Pediatric asthma: Principles and treatment. Allergy Asthma Proc. 2019;40(6):389–392. 10.2500/aap.2019.40.4254
4. Cloutier MM, Teach SJ, Lemanske RF, Jr., Blake KV. The 2020 focused updates to the NIH Asthma Management Guidelines: Key points for pediatricians. Pediatrics. 2021;147(6): e2021050286. 10.1542/peds.2021-050286
5. Serebrisky D, Wiznia A. Pediatric asthma: A global epidemic. Ann Glob Health. 2019;85(1):6. 10.5334/aogh.2416
6. Akar-Ghibril N, Casale T, Custovic A, Phipatanakul W. Allergic endotypes and phenotypes of asthma. J Allergy Clin Immunol Pract. 2020;8(2):429–440. 10.1016/j.jaip.2019.11.008
7. Wechsler ME, Munitz A, Ackerman SJ, Drake MG, Jackson DJ, Wardlaw AJ, et al. Eosinophils in health and disease: A state-of-the-art review. Mayo Clin Proc. 2021;96(10):2694–2707. 10.1016/j.mayocp.2021.04.025
8. Van Hulst G, Bureau F, Desmet CJ. Eosinophils as drivers of severe eosinophilic asthma: Endotypes or plasticity? Int J Mol Sci. 2021;22(18):10150. 10.3390/ijms221810150
9. Dunn JLM, Rothenberg ME. 2021 year in review: Spotlight on eosinophils. J Allergy Clin Immunol. 2022;149(2):517–524. 10.1016/j.jaci.2021.11.012
10. Aoki A, Hirahara K, Kiuchi M, Nakayama T. Eosinophils: Cells known for over 140 years with broad and new functions. Allergol Int. 2021;70(1):3–8. 10.1016/j.alit.2020.09.002
11. Felton JM, Dorward DA, Cartwright JA, Potey PM, Robb CT, Gui J, et al. Mcl-1 protects eosinophils from apoptosis and exacerbates allergic airway inflammation. Thorax. 2020;75(7):600–605. 10.1136/thoraxjnl-2019-213204
12. Li J, Cao F, Yin HL, Huang ZJ, Lin ZT, Mao N, et al. Ferroptosis: past, present and future. Cell Death Dis. 2020;11(2):88. 10.1038/s41419-023-05595-5. 10.1038/s41419-020-2298-2
13. Wu Y, Chen H, Xuan N, Zhou L, Wu Y, Zhu C, et al. Induction of ferroptosis-like cell death of eosinophils exerts synergistic effects with glucocorticoids in allergic airway inflammation. Thorax. 2020;75(11):918–927. 10.1136/thoraxjnl-2020-214764
14. Ora J, Calzetta L, Matera MG, Cazzola M, Rogliani P. Advances with glucocorticoids in the treatment of asthma: State of the art. Expert Opin Pharmacother. 2020;21(18):2305–2316. 10.1080/14656566.2020.1807514
15. Enweasor C, Flayer CH, Haczku A. Ozone-induced oxidative stress, neutrophilic airway inflammation, and glucocorticoid resistance in asthma. Front Immunol. 2021;12:631092. 10.3389/fimmu.2021.631092
16. Poddighe D, Brambilla I, Licari A, Marseglia GL. Omalizumab in the therapy of pediatric asthma. Recent Pat Inflamm Allergy Drug Discov. 2018;12(2):103–109. 10.2174/1872213X12666180430161351
17. Steven A, Friedrich M, Jank P, Heimer N, Budczies J, Denkert C, et al. What turns CREB on? And off? And why does it matter? Cell Mol Life Sci. 2020;77(20):4049–4067. 10.1007/s00018-020-03525-8
18. Wang Z, Zhang X, Tian X, Yang Y, Ma L, Wang J, et al. CREB stimulates GPX4 transcription to inhibit ferroptosis in lung adenocarcinoma. Oncol Rep. 2021;45(6):88. 10.3892/or.2021.8039
19. Wang H, Xu J, Lazarovici P, Quirion R, Zheng W. cAMP response element-binding protein (CREB): A possible signaling molecule link in the pathophysiology of schizophrenia. Front Mol Neurosci. 2018;11:255. 10.3389/fnmol.2018.00255
20. Chen X, Li XM, Gu W, Wang D, Chen Y, Guo XJ. LAT alleviates Th2/Treg imbalance in an OVA-induced allergic asthma mouse model through LAT-PLC-gamma1 interaction. Int Immunopharmacol. 2017;44:9–15. 10.1016/j.intimp.2016.12.029
21. National Research Council (US) Committee for the Update of the Guide for the Care and Use of Laboratory Animals. Guide for the Care and Use of Laboratory Animals. 8th ed. Washington, DC: National Academy of Sciences, National Academies Press (US); 2011. The National Academies Collection: Reports funded by National Institutes of Health.
22. Rahmati M, Shariatzadeh Joneydi M, Koyanagi A, Yang G, Ji B, Won Lee S, et al. Resistance training restores skeletal muscle atrophy and satellite cell content in an animal model of Alzheimer’s disease. Sci Rep. 2023;13(1):2535. 10.1038/s41598-023-29406-1
23. Rahmati M, Taherabadi SJ. The effects of exercise training on Kinesin and GAP-43 expression in skeletal muscle fibers of STZ-induced diabetic rats. Sci Rep. 2021;11(1):9535. 10.1038/s41598-021-89106-6
24. Yu QL, Chen Z. Establishment of different experimental asthma models in mice. Exp Ther Med. 2018;15(3):2492–2498. 10.3892/etm.2018.5721
25. Bostani M, Rahmati M, Mard SA. The effect of endurance training on levels of LINC complex proteins in skeletal muscle fibers of STZ-induced diabetic rats. Sci Rep. 2020;10(1):8738. 10.1038/s41598-020-65793-5
26. Dong L, Wang Y, Zheng T, Pu Y, Ma Y, Qi X, et al. Hypoxic hUCMSC-derived extracellular vesicles attenuate allergic airway inflammation and airway remodeling in chronic asthma mice. Stem Cell Res Ther. 2021;12(1):4. 10.1186/s13287-020-02072-0
27. Azmeh R, Greydanus DE, Agana MG, Dickson CA, Patel DR, Ischander MM, et al. Update in pediatric asthma: Selected issues. Dis Mon. 2020;66(4):100886. 10.1016/j.disamonth.2019.100886
28. Liu L, Ma H, Yuan S, Zhang J, Wu J, Dilimulati M, et al. Prognosis of bronchial asthma in children with different pulmonary function phenotypes: A real-world retrospective observational study. Front Pediatr. 2022;10:1043047. 10.3389/fped.2022.1043047
29. Wu HK, Chang ES, Cheng CW, Chien WC, Chou HH. Impact of COVID-19 pandemic waves on pediatric emergency department patients presenting with asthma attacks in Taiwan. Signa Vitae. 2022; 18(6):27-32. https://www.signavitae.com/articles/10.22514/sv.2022.045
30. Tanaka K, Arakawa M, Miyake Y. Perinatal smoking exposure and risk of asthma in the first three years of life: A prospective prebirth cohort study. Allergol Immunopathol (Madr). 2020;48(6):530–536. 10.1016/j.aller.2020.03.008
31. Bimstein E, Wilson J, Guelmann M, Primosch RE. The relationship between oral and demographic characteristics of children with asthma. J Clin Pediatr Dent. 2006;31(2):86–89. 10.17796/jcpd.31.2.977776084511l005
32. Funston W, Higgins B. Improving the management of asthma in adults in primary care. Practitioner. 2014;258(1776):15–19.
33. Pelaia C, Crimi C, Vatrella A, Tinello C, Terracciano R, Pelaia G. Molecular targets for biological therapies of severe asthma. Front Immunol. 2020;11:603312. 10.3389/fimmu.2020.603312
34. Brussino L, Heffler E, Bucca C, Nicola S, Rolla G. Eosinophils target therapy for severe asthma: Critical points. Biomed Res Int. 2018;2018:7582057. 10.1155/2018/7582057
35. Jacobsen EA, Jackson DJ, Heffler E, Mathur SK, Bredenoord AJ, Pavord ID, et al. Eosinophil knockout humans: Uncovering the role of eosinophils through eosinophil-directed biological therapies. Annu Rev Immunol. 2021;39:719–757. 10.1146/annurev-immunol-093019-125918
36. Ilmarinen P, Kankaanranta H. Eosinophil apoptosis as a therapeutic target in allergic asthma. Basic Clin Pharmacol Toxicol. 2014;114(1):109–117. 10.1111/bcpt.12163
37. Zirnask H, Pöllanen P, Suutre S, Kuuslahti M, Kotsar A, Pakarainen T, et al. Expression of cAMP and CREB in the human penis. J Men’s Health. 2019;15(4):12–17.
38. Cui P, Shi K, Cui HX, Hao LY, Su Y, Li PL. Correlation between transcription factor activator protein-2β (TFAP-2β) and endometrial carcinoma. Eur J Gynaecol Oncol. 2015;36(3):268–273.
39. Koopmans T, Crutzen S, Menzen MH, Halayko AJ, Hackett TL, Knight DA, et al. Selective targeting of CREB-binding protein/beta-catenin inhibits growth of and extracellular matrix remodelling by airway smooth muscle. Br J Pharmacol. 2016;173(23):3327–3341. 10.1111/bph.13620
40. Koga Y, Hisada T, Ishizuka T, Utsugi M, Ono A, Yatomi M, et al. CREB regulates TNF-alpha-induced GM-CSF secretion via p38 MAPK in human lung fibroblasts. Allergol Int. 2016;65(4):406–413. 10.1016/j.alit.2016.03.006
41. Wu Y, Lu Y, Zou F, Fan X, Li X, Zhang H, et al. PTEN participates in airway remodeling of asthma by regulating CD38/Ca(2+)/CREB signaling. Aging (Albany NY). 2020;12(16):16326–16340. 10.18632/aging.103664
42. Galaris D, Barbouti A, Pantopoulos K. Iron homeostasis and oxidative stress: An intimate relationship. Biochim Biophys Acta Mol Cell Res. 2019;1866(12):118535. 10.1016/j.bbamcr.2019.118535
43. Chen K, Xue R, Geng Y, Zhang S. Galangin inhibited ferroptosis through activation of the PI3K/AKT pathway in vitro and in vivo. FASEB J. 2022;36(11):e22569. 10.1096/fj.202200935R
44. Xiong Q, Tian X, Xu C, Ma B, Liu W, Sun B, et al. PM(2) (.5) exposure-induced ferroptosis in neuronal cells via inhibiting ERK/CREB pathway. Environ Toxicol. 2022;37(9):2201–2213. 10.1002/tox.23586
45. Hu M, Zhong Y, Liu J, Zheng S, Lin L, Lin X, et al. An adverse outcome pathway-based approach to assess aurantio-obtusin-induced hepatotoxicity. Toxicology. 2022;478:153293. 10.1016/j.tox.2022.153293
Article Sidebar
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.