Mathies M.a· Krieg E.M.a· Mohr F.a· Zaradzki M.b· Wagner A.H.a

Author affiliations

aDepartment of Cardiovascular Physiology, Heidelberg University, Heidelberg, Germany
bDepartment of Cardiac Surgery, University Hospital Heidelberg, Heidelberg, Germany

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Article / Publication Details

First-Page Preview

Received: March 29, 2022
Accepted: August 09, 2022
Published online: September 29, 2022

Number of Print Pages: 8
Number of Figures: 4
Number of Tables: 0

ISSN: 0031-7012 (Print)
eISSN: 1423-0313 (Online)

For additional information: https://www.karger.com/PHA

Abstract

Activation of the mechanistic target of rapamycin (mTOR) pathway has been implicated in an increasing number of diseases, including Marfan syndrome (MFS), an inherited connective tissue disorder. mTOR-dependent reactive oxygen species (ROS) formation has also been suggested to play a role in aortic aneurysm formation in MFS patients. This study aimed to characterize the effects of mTOR inhibition by rapamycin on key redox enzymes and NADPH oxidases (NOX) in cultured vascular smooth muscle cells of a murine MFS model. Therefore, the influence of 5 and 20 nmol/L rapamycin solved in 0.1% (vol/vol) DMSO on glutathione peroxidases 1 (Gpx1) and 4 (Gpx4), superoxide dismutase 2 (Sod2), and catalase (Cat) mRNA and protein expression was investigated in isolated murine aortic smooth muscle cells. Rapamycin inhibited the mRNA expression of all redox enzymes by 30–50%, except Gpx1. In the same cells, the mRNA expression of the transcription factor NFE2-related factor-2 and peroxisome proliferator-activated receptor-γ, key factors against oxidative stress, and controlling redox gene expression were also inhibited to a comparable extent under these conditions. In addition, Nox1 but not Nox4 mRNA expression was significantly inhibited by up to 40%. DMSO alone increased nearly 2-fold the redox enzyme protein expression, which was reduced considerably to basal levels by rapamycin. Proteasomal inhibition by bortezomib could not reverse the observed decrease of GPx protein content. The rapamycin-mediated decrease in GPx protein abundance was reflected in a reduced total GPx enzymatic activity. Higher rapamycin concentrations did not further decrease but led to a renewed increase in enzymatic activity despite low GPx protein concentrations. Baseline ROS formation was slightly inhibited at 13% with 5 nmol/L rapamycin and returned to baseline levels with the higher 20 nmol/L rapamycin concentration. In conclusion, this study further characterized the mechanism of action of rapamycin. It provided an insight into how rapamycin interferes with the regulation of redox homeostasis essential for ROS-dependent signaling that does not incur cellular damage.

© 2022 S. Karger AG, Basel

References Wagner AH, Zaradzki M, Arif R, Remes A, Müller OJ, Kallenbach K. Marfan syndrome: a therapeutic challenge for long-term care. Biochem Pharmacol. 2019 Jun;164:53–63. Barrett PM, Topol EJ. The fibrillin-1 gene: unlocking new therapeutic pathways in cardiovascular disease. Heart. 2013 Jan;99(2):83–90. Judge DP, Dietz HC. Therapy of Marfan syndrome. Annu Rev Med. 2008;59:43–59. Li J, Kim SG, Blenis J. Rapamycin: one drug, many effects. Cell Metab. 2014;19(3):373–9. Li G, Wang M, Caulk AW, Cilfone NA, Gujja S, Qin L, et al. Chronic mTOR activation induces a degradative smooth muscle cell phenotype. J Clin Invest. 2020 Mar 2;130(3):1233–51. Cao J, Wu Q, Geng L, Chen X, Shen W, Wu F, et al. Rapamycin inhibits CaCl2-induced thoracic aortic aneurysm formation in rats through mTOR-mediated suppression of proinflammatory mediators. Mol Med Rep. 2017 Aug;16(2):1911–9. Nguyen LS, Vautier M, Allenbach Y, Zahr N, Benveniste O, Funck-Brentano C, et al. Sirolimus and mTOR inhibitors: a review of side effects and specific management in solid organ transplantation. Drug Saf. 2019 Jul;42(7):813–25. Jiménez-Altayó F, Meirelles T, Crosas-Molist E, Sorolla MA, Del Blanco DG, López-Luque J, et al. Redox stress in Marfan syndrome: dissecting the role of the NADPH oxidase NOX4 in aortic aneurysm. Free Radic Biol Med. 2018 Apr;118:44–58. Rysz J, Gluba-Brzózka A, Rokicki R, Franczyk B. Oxidative stress-related susceptibility to aneurysm in Marfan’s syndrome. Biomedicines. 2021 Sep 6;9(9):1171. Schwill S, Seppelt P, Grünhagen J, Ott C-E, Jugold M, Ruhparwar A, et al. The fibrillin-1 hypomorphic mgR/mgR murine model of Marfan syndrome shows severe elastolysis in all segments of the aorta. J Vasc Surg. 2013 Jan;57(6):1628–36, 1636.e1–3. Arif R, Franz M, Remes A, Zaradzki M, Hecker M, Karck M, et al. Reduction of transplant vasculopathy by intraoperative nucleic acid-based therapy in a mouse aortic allograft model. Thorac Cardiovasc Surg. 2019 Sep;67(6):503–12. Wagner AH, Güldenzoph B, Lienenlüke B, Hecker M. CD154/CD40-mediated expression of CD154 in endothelial cells: consequences for endothelial cell-monocyte interaction. Arterioscler Thromb Vasc Biol. 2004 Apr;24(4):715–20. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001 May 1;29(9):e45. Sultan CS, Saackel A, Stank A, Fleming T, Fedorova M, Hoffmann R, et al. Impact of carbonylation on glutathione peroxidase-1 activity in human hyperglycemic endothelial cells. Redox Biol. 2018 May;16:113–22. DiJoseph JF, Fluhler E, Armstrong J, Sharr M, Sehgal SN. Therapeutic blood levels of sirolimus (rapamycin) in the allografted rat. Transplantation. 1996 Oct 27;62(8):1109–12. Dovinova I, Kvandová M, Balis P, Gresova L, Majzunova M, Horakova L, et al. The role of Nrf2 and PPARγ in the improvement of oxidative stress in hypertension and cardiovascular diseases. Physiol Res. 2020 Dec 31;69(Suppl 4):S541–53. Zhao J, Zhai B, Gygi SP, Goldberg AL. mTOR inhibition activates overall protein degradation by the ubiquitin proteasome system as well as by autophagy. Proc Natl Acad Sci U S A. 2015 Dec;112(52):15790–7. Adams J, Kauffman M. Development of the proteasome inhibitor Velcade (Bortezomib). Cancer Invest. 2004;22(2):304–11. Durgin BG, Straub AC. Redox control of vascular smooth muscle cell function and plasticity. Lab Invest. 2018 Oct;98(10):1254–62. Reinke EN, Ekoue DN, Bera S, Mahmud N, Diamond AM. Translational regulation of GPx-1 and GPx-4 by the mTOR pathway. PLoS One. 2014;9(4):e93472. Yoo Y-M, Jung E-M, Jeung E-B. Rapamycin-induced autophagy decreases Myf5 and MyoD proteins in C2C12 myoblast cells. Toxicol In Vitro. 2019 Aug;58:132–41. Zhang Y, Swanda RV, Nie L, Liu X, Wang C, Lee H, et al. mTORC1 couples cyst(e)ine availability with GPX4 protein synthesis and ferroptosis regulation. Nat Commun. 2021 Mar 11;12(1):1589. Reho JJ, Guo DF, Rahmouni K. Mechanistic target of rapamycin complex 1 signaling modulates vascular endothelial function through reactive oxygen species. J Am Heart Assoc. 2019;8(9):e010662. Tan P, Wang YJ, Li S, Wang Y, He JY, Chen YY, et al. The PI3K/Akt/mTOR pathway regulates the replicative senescence of human VSMCs. Mol Cell Biochem. 2016 Nov;422(1–2):1–10. Saxton RA, Sabatini DM. mTOR signaling in growth, metabolism, and disease. Cell. 2017;169(2):361–71. Kim JE, Chen J. Regulation of peroxisome proliferator-activated receptor-gamma activity by mammalian target of rapamycin and amino acids in adipogenesis. Diabetes. 2004 Nov;53(11):2748–56. Glorieux C, Auquier J, Dejeans N, Sid B, Demoulin JB, Bertrand L, et al. Catalase expression in MCF-7 breast cancer cells is mainly controlled by PI3K/Akt/mTor signaling pathway. Biochem Pharmacol. 2014 May 15;89(2):217–23. Woo Y, Lee HJ, Jung YM, Jung YJ. mTOR-mediated antioxidant activation in solid tumor radioresistance. J Oncol. 2019;2019:5956867. Nieradka A, Ufer C, Thiadens K, Grech G, Horos R, van Coevorden-Hameete M, et al. Grsf1-induced translation of the SNARE protein Use1 is required for expansion of the erythroid compartment. PLoS One. 2014;9(9):e104631. Ufer C, Wang CC, Fähling M, Schiebel H, Thiele BJ, Billett EE, et al. Translational regulation of glutathione peroxidase 4 expression through guanine-rich sequence-binding factor 1 is essential for embryonic brain development. Genes Dev. 2008 Jul 1;22(13):1838–50. Martin KA, Rzucidlo EM, Merenick BL, Fingar DC, Brown DJ, Wagner RJ, et al. The mTOR/p70 S6K1 pathway regulates vascular smooth muscle cell differentiation. Am J Physiol Cell Physiol. 2004 Mar;286(3):C507–17. Cengiz Seval G, Beksac M. The safety of bortezomib for the treatment of multiple myeloma. Expert Opin Drug Saf. 2018 Sep;17(9):953–62. Ellmark SHM, Dusting GJ, Fui MNT, Guzzo-Pernell N, Drummond GR. The contribution of Nox4 to NADPH oxidase activity in mouse vascular smooth muscle. Cardiovasc Res. 2005 Feb 1;65(2):495–504. Clempus RE, Sorescu D, Dikalova AE, Pounkova L, Jo P, Sorescu GP, et al. Nox4 is required for maintenance of the differentiated vascular smooth muscle cell phenotype. Arterioscler Thromb Vasc Biol. 2007 Jan;27(1):42–8. Fernandes DC, Wosniak J Jr, Gonçalves RC, Tanaka LY, Fernandes CG, Zanatta DB, et al. PDIA1 acts as master organizer of NOX1/NOX4 balance and phenotype response in vascular smooth muscle. Free Radic Biol Med. 2021 Jan;162:603–14. Li W, Li Q, Qin L, Ali R, Qyang Y, Tassabehji M, et al. Rapamycin inhibits smooth muscle cell proliferation and obstructive arteriopathy attributable to elastin deficiency. Arterioscler Thromb Vasc Biol. 2013 May;33(5):1028–35. Huang WQ, Zou Y, Tian Y, Ma XF, Zhou QY, Li ZY, et al. Mammalian target of rapamycin as the therapeutic target of vascular proliferative diseases: past, present, and future. J Cardiovasc Pharmacol. 2022 Apr 1;79(4):444–55. Cattaruzza M, Hecker M. Protein carbonylation and decarboylation: a new twist to the complex response of vascular cells to oxidative stress. Circ Res. 2008 Feb 15;102(3):273–4. Werner E. Determination of cellular H2O2 production. Sci STKE. 2003 Feb 4;2003(168):PL3. Article / Publication Details

First-Page Preview

Received: March 29, 2022
Accepted: August 09, 2022
Published online: September 29, 2022

Number of Print Pages: 8
Number of Figures: 4
Number of Tables: 0

ISSN: 0031-7012 (Print)
eISSN: 1423-0313 (Online)

For additional information: https://www.karger.com/PHA

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