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Rapamycin Is Not Protective against Ischemic and Cisplatin-Induced Kidney Injury

Abstract

Autophagy plays an important role in the pathogenesis of acute kidney injury (AKI). Although autophagy activation was shown to be associated with an increased lifespan and beneficial effects in various pathologies, the impact of autophagy activators, particularly, rapamycin and its analogues on AKI remains obscure. In our study, we explored the effects of rapamycin treatment in in vivo and in vitro models of ischemic and cisplatin-induced AKI. The impact of rapamycin on the kidney function after renal ischemia/reperfusion (I/R) or exposure to the nephrotoxic agent cisplatin was assessed by quantifying blood urea nitrogen and serum creatinine and evaluating the content of neutrophil gelatinase-associated lipocalin, a novel biomarker of AKI. In vitro experiments were performed on the primary culture of renal tubular cells (RTCs) that were subjected to oxygen-glucose deprivation (OGD) or incubated with cisplatin under various rapamycin treatment protocols. Cell viability and proliferation were estimated by the MTT assay and real-time cell analysis using an RTCA iCELLigence system. Although rapamycin inhibited mTOR (mammalian target of rapamycin) signaling, it failed to enhance the autophagy and to ameliorate the severity of AKI caused by ischemia or cisplatin-induced nephrotoxicity. Experiments with RTCs demonstrated that rapamycin exhibited the anti-proliferative effect in primary RTCs cultures but did not protect renal cells exposed to OGD or cisplatin. Our study revealed for the first time that the mTOR inhibitor rapamycin did not prevent AKI caused by renal I/R or cisplatin-induced nephrotoxicity and, therefore, cannot be considered as an ideal mimetic of the autophagy-associated nephroprotective mechanisms (e.g., those induced by caloric restriction), as it had been suggested earlier. The protective action of such approaches like caloric restriction might not be limited to mTOR inhibition and can proceed through more complex mechanisms involving alternative autophagy-related targets. Thus, the use of rapamycin and its analogues for the treatment of various AKI forms requires further studies in order to understand potential protective or adverse effects of these compounds in different contexts.

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Abbreviations

AKI:

acute kidney injury

BUN:

blood urea nitrogen

DMSO:

dimethyl sulfoxide

DPBS:

Dulbecco’s phosphatebuffered saline

EGF:

epidermal growth factor

FBS:

fetal bovine serum; i.p., intraperitoneal (injection)

I/R:

ischemia/reperfusion

LC3:

microtubule-associated protein 1A/1B, light chain 3

mTOR:

mammalian target of rapamycin

mTORC1:

mammalian target of rapamycin complex 1

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium

NGAL:

neutrophil gelatinase-associated lipocalin

OGD:

oxygen-glucose deprivation

RTCs:

renal tubular cells

SCr:

serum creatinine

References

  1. 1.

    Levine, B., and Kroemer, G. (2008) Autophagy in the pathogenesis of disease, Cell, 132, 27–42, doi: 10.1016/j.cell.2007.12.018.

  2. 2.

    Mizushima, N., Levine, B., Cuervo, A. M., and Klionsky, D. J. (2008) Autophagy fights disease through cellular selfdigestion, Nature, 451, 1069–1075, doi: 10.1038/nature06639.

  3. 3.

    Ryu, D., Mouchiroud, L., Andreux, P. A., Katsyuba, E., Moullan, N., Nicolet-Dit-Felix, A. A., Williams, E. G., Jha, P., Lo Sasso, G., Huzard, D., Aebischer, P., Sandi, C., Rinsch, C., and Auwerx, J. (2016) Urolithin A induces mitophagy and prolongs lifespan in C. elegans and increases muscle function in rodents, Nat. Med., 22, 879–888, doi: 10.1038/nm.4132.

  4. 4.

    Andreux, P. A., Blanco-Bose, W., Ryu, D., Burdet, F., Ibberson, M., Aebischer, P., Auwerx, J., Singh, A., and Rinsch, C. (2019) The mitophagy activator urolithin A is safe and induces a molecular signature of improved mitochondrial and cellular health in humans, Nat. Metab., 1, 595–603, doi: 10.1038/s42255-019-0073-4.

  5. 5.

    Periyasamy-Thandavan, S., Jiang, M., Wei, Q., Smith, R., Yin, X.-M., and Dong, Z. (2008) Autophagy is cytoprotective during cisplatin injury of renal proximal tubular cells, Kidney Int., 1, 631–640, doi: 10.1038/ki.2008.214.

  6. 6.

    Takahashi, A., Kimura, T., Takabatake, Y., Namba, T., Kaimori, J., Kitamura, H., Matsui, I., Niimura, F., Matsusaka, T., Fujita, N., Yoshimori, T., Isaka, Y., and Rakugi, H. (2012) Autophagy guards against cisplatininduced acute kidney injury, Am. J. Pathol., 1, 517–525, doi: 10.1016/j.ajpath.2011.11.001.

  7. 7.

    Liu, S., Hartleben, B., Kretz, O., Wiech, T., Igarashi, P., Mizushima, N., Walz, G., and Huber, T. B. (2012) Autophagy plays a critical role in kidney tubule maintenance, aging and ischemia-reperfusion injury, Autophagy, 8, 826–837, doi: 10.4161/auto.19419.

  8. 8.

    Li, L., Wang, Z. V., Hill, J. A., and Lin, F. (2014) New autophagy reporter mice reveal dynamics of proximal tubular autophagy, J. Am. Soc. Nephrol., 1, 305–315, doi: 10.1681/ASN.2013040374.

  9. 9.

    Li, T., Liu, Y., Zhao, J., Miao, S., Xu, Y., and Liu, K., Liu, M., Wang, G., and Xiao, X. (2017) Aggravation of acute kidney injury by mPGES-2 down regulation is associated with autophagy inhibition and enhanced apoptosis, Sci. Rep., 7, 10247, doi: 10.1038/s41598-017-10271-8.

  10. 10.

    Mei, S., Livingston, M., Hao, J., Li, L., Mei, C., and Dong, Z. (2016) Autophagy is activated to protect against endotoxic acute kidney injury, Sci. Rep., 6, 22171, doi: 10.1038/srep22171.

  11. 11.

    Karagiannidis, I., Kataki, A., Glustianou, G., Memos, N., Papalois, A., and Alexakis, N., Zografos, G. C., and Konstadoulakis, M. M. (2016) Extended cytoprotective effect of autophagy in the late stages of sepsis and fluctuations in signal transduction pathways in a rat experimental model of kidney injury, Shock, 45,139-147, doi: 10.1097/SHK.0000000000000505.

  12. 12.

    Ko, G. J., Bae, S. Y., Hong, Y.-A., Pyo, H. J., and Kwon, Y. J. (2016) Radiocontrast-induced nephropathy is attenuated by autophagy through regulation of apoptosis and inflammation, Hum. Exp. Toxicol., 1, 724–736, doi: 10.1177/0960327115604198.

  13. 13.

    Kimura, T., Takabatake, Y., Takahashi, A., Kaimori, J., Matsui, I., Namba, T., Kitamura, H., Niimura, F., Matsusaka, T., Soga, T., Rakugi, H., and Isaka, Y. (2011) Autophagy protects the proximal tubule from degeneration and acute ischemic injury, J. Am. Soc. Nephrol., 1, 902–913, doi: 10.1681/ASN.2010070705.

  14. 14.

    Jiang, M., Wei, Q., Dong, G., Komatsu, M., Su, Y., and Dong, Z. (2012) Autophagy in proximal tubules protects against acute kidney injury, Kidney Int., 1, 1271–1283, doi: 10.1038/ki.2012.261.

  15. 15.

    Andrianova, N. V., Jankauskas, S. S., Zorova, L. D., Pevzner, I. B., Popkov, V. A., Silachev, D. N., Plotnikov, E. Y., and Zorov, D. B. (2018) Mechanisms of age-dependent loss of dietary restriction protective effects in acute kidney injury, Cells, 7, 178, doi: 10.3390/cells7100178.

  16. 16.

    Kuo, S.-Y., Castoreno, A. B., Aldrich, L. N., Lassen, K. G., Goel, G., Dancik, V., Kuballa, P., Latorre, I., Conway, K. L., Sarkar, S., Maetzel, D., Jaenisch, R., Clemons, P. A., Schreiber, S. L., Shamji, A. F., and Xavier, R. J. (2015) Smallmolecule enhancers of autophagy modulate cellular disease phenotypes suggested by human genetics, Proc. Natl. Acad. Sci. USA, 112, 4281–4287, doi: 10.1073/pnas.1512289112.

  17. 17.

    Blagosklonny, M. V. (2017) From rapalogs to anti-aging formula, Oncotarget, 8, 35492–35507, doi: 10.18632/oncotarget.18033.

  18. 18.

    Plotnikov, E. Y., Kazachenko, A. V., Vyssokikh, M. Y., Vasileva, A. K., Tcvirkun, D. V., Isaev, N. K., Kirpatovsky, V. I., and Zorov, D. B. (2007) The role of mitochondria in oxidative and nitrosative stress during ischemia/reperfusion in the rat kidney, Kidney Int., 1, 1493–502, doi: 10.1038/sj.ki.5002568.

  19. 19.

    Schneider, C. A., Rasband, W. S., and Eliceiri, K. W. (2012) NIH image to imageJ: 25 years of image analysis, Nat. Methods, 9, 671–675.

  20. 20.

    Giaever, I., and Keese, C. R. (1984) Monitoring fibroblast behavior in tissue culture with an applied electric field, Proc. Natl. Acad. Sci. USA, 81, 3761–3764.

  21. 21.

    Popkov, V. A., Andrianova, N. V., Manskikh, V. N., Silachev, D. N., Pevzner, I. B., and Zorova, L. D., Sukhikh, G. T., Plotnikov, E. Y., and Zorov, D. B. (2018) Pregnancy protects the kidney from acute ischemic injury, Sci. Rep., 8, 14534, doi: 10.1038/s41598-018-32801-8.

  22. 22.

    Tavares, M. R., Pavan, C. B., Amaral, C. L., Meneguello, L., Luchessi, A. D., and Simabuco, F. M. (2015) The S6K protein family in health and disease, Life Sci., 1, 1–10, doi: 10.1016/j.lfs.2015.03.001.

  23. 23.

    Stankov, M., Panayotova-Dimitrova, D., Leverkus, M., Klusmann, J.-H., and Behrens, G. (2014) Cytometric analysis of autophagic activity with cyto-ID staining in primary cells, Bio-Protocol, 4, doi: 10.21769/BioProtoc.1090.

  24. 24.

    Todorovic, Z., Medic, B., Basta-Jovanovic, G., Radojevic Skodric, S., Stojanovic, R., Rovcanin, B., and Prostran, M. (2014) Acute pretreatment with chloroquine attenuates renal I/R injury in rats, PLoS One, 9, 92673, doi: 10.1371/journal.pone.0092673.

  25. 25.

    Zhang, Y.-L., Zhang, J., Cui, L.-Y., and Yang, S. (2015) Autophagy activation attenuates renal ischemia-reperfusion injury in rats, Exp. Biol. Med., 1, 1590–1598, doi: 10.1177/1535370215581306.

  26. 26.

    Guan, X., Qian, Y., Shen, Y., Zhang, L., Du, Y., Dai, H., Qian, J., and Yan, Y. (2015) Autophagy protects renal tubular cells against ischemia/reperfusion injury in a timedependent manner, Cell Physiol. Biochem., 1, 285–298, doi: 10.1159/000374071.

  27. 27.

    Ling, H., Chen, H., Wei, M., Meng, X., Yu, Y., and Xie, K. (2016) The effect of autophagy on inflammation cytokines in renal ischemia/reperfusion injury, Inflammation, 39, 347–356, doi: 10.1007/s10753-015-0255-5.

  28. 28.

    Zhu, J., Lu, T., Yue, S., Shen, X., Gao, F., Busuttil, R. W., Kupiec-Weglinski, J. W., Xia, Q., and Zhai, Y. (2015) Rapamycin protection of livers from ischemia and reperfusion injury is dependent on both autophagy induction and mammalian target of rapamycin complex 2-Akt activation, Transplantation, 99, 48–55, doi: 10.1097/TP.0000000000000476.

  29. 29.

    Parra, C., Salas, P., and Dominguez, J. (2010) Effects of immunosuppressive drugs on rat renal ischemia reperfusion injury, Transplant. Proc., 1, 245–247, doi: 10.1016/j.transproceed.2009.11.018.

  30. 30.

    Pereira, B. J., Castro, I., Burdmann, E. A., Malheiros, D. M. A., and Yu, L. (2010) Effects of sirolimus alone or in combination with cyclosporine A on renal ischemia/reperfusion injury, Braz. J. Med. Biol. Res., 43, 737–744.

  31. 31.

    Goncalves, G. M., Cenedeze, M. A., Feitoza, C. Q., de Paula, C. B., Marques, G. D., Pinheiro, H. S., de Paula Antunes Teixeira, V., Antonia dos Reis, M., Pacheco-Silva, A., and Camara, N. O. (2007) The role of immunosuppressive drugs in aggravating renal ischemia and reperfusion injury, Transplant. Proc., 1, 417–420, doi: 10.1016/j.transproceed.2007.01.027.

  32. 32.

    Lui, S. L., Chan, K. W., Tsang, R., Yung, S., Lai, K. N., and Chan, T. M. (2006) Effect of rapamycin on renal ischemia-reperfusion injury in mice, Transpl. Int., 1, 834–839, doi: 10.1111/j.1432-2277.2006.00361.x.

  33. 33.

    Lieberthal, W., Fuhro, R., Andry, C. C., Rennke, H., Abernathy, V. E., Koh, J. S., Valeri, R., and Levine, J. S. (2001) Rapamycin impairs recovery from acute renal failure: role of cell-cycle arrest and apoptosis of tubular cells, Am. J. Physiol. Renal Physiol., 1, 693–706, doi: 10.1152/ajprenal.2001.281.4.F693.

  34. 34.

    Martel, R. R., Klicius, J., and Galet, S. (1977) Inhibition of the immune response by rapamycin, a new antifungal antibiotic, Can. J. Physiol. Pharmacol., 1, 48–51.

  35. 35.

    Kim, B. S., Lim, S. W., Li, C., Kim, J. S., Sun, B. K., Ahn, K. O., Han, S. W., Kim, J., and Yang, C. W. (2005) Ischemia-reperfusion injury activates innate immunity in rat kidneys, Transplantation, 79, 1370–1377.

  36. 36.

    Dumont, F. J., and Su, Q. (1996) Mechanism of action of the immunosuppressant rapamycin, Life Sci., 1, 373–395.

  37. 37.

    Law, B. K. (2005) Rapamycin: an anti-cancer immunosuppressant? Crit. Rev. Oncol. Hematol., 56, 47–60, doi: 10.1016/j.critrevonc.2004.09.009.

  38. 38.

    Kaushal, G. P. (2012) Autophagy protects proximal tubular cells from injury and apoptosis, Kidney Int., 1, 1250–1253, doi: 10.1038/ki.2012.337.

  39. 39.

    Nakagawa, S., Nishihara, K., Inui, K., and Masuda, S. (2012) Involvement of autophagy in the pharmacological effects of the mTOR inhibitor everolimus in acute kidney injury, Eur. J. Pharmacol., 1, 43–54, doi: 10.1016/j.ejphar.2012.09.010.

  40. 40.

    Mitchell, J. R., Verweij, M., Brand, K., van de Ven, M., Goemaere, N., van den Engel, S., Chu, T., Forrer, F., Muller, C., de Jong, M., van IJcken, W., IJzermans, J. N., Hoeijmakers, J. H., and de Bruin, R. W. (2010) Short-term dietary restriction and fasting precondition against ischemia reperfusion injury in mice, Aging Cell, 9, 40–53, doi: 10.1111/j.1474-9726.2009.00532.x.

  41. 41.

    Ning, Y.-C., Cai, G.-Y., Zhuo, L., Gao, J. J., Dong, D., Cui, S.-Y., Shi, S. Z., Feng, Z., Zhang, L., Sun, X. F., and Chen, X. M. (2013) Beneficial effects of short-term calorie restriction against cisplatin-induced acute renal injury in aged rats, Nephron. Exp. Nephrol., 1, 9–27, doi: 10.1159/000357380.

  42. 42.

    Tokunaga, C., Yoshino, K., and Yoneezawa, K. (2004) mTOR integrates amino acidand energy-sensing pathways, Biochem. Biophys. Res. Commun., 1, 443–446.

  43. 43.

    Li, J., Kim, S. G., and Blenis, J. (2014) Rapamycin: one drug, many effects, Cell Metab., 1, 373–379, doi: 10.1016/j.cmet.2014.01.001.

  44. 44.

    Lee, S.-H., and Min, K.-J. (2013) Caloric restriction and its mimetics, BMB Rep., 1, 181–187.

  45. 45.

    Lamming, D. W. (2016) Inhibition of the mechanistic target of rapamycin (mTOR) - rapamycin and beyond, Cold Spring Harb. Perspect. Med.,6, a025924, doi: 10.1101/cshperspect.a025924.

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Correspondence to E. Y. Plotnikov.

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Conflict of interest. The authors declare no conflict of interest.

Compliance with ethical norms. The rats were treated according to the protocols evaluated and approved by the Animal Ethics Committee of Belozersky Institute of Physico-Chemical Biology. All procedures were in accordance with the guidelines of the Federation of Laboratory Animal Science Associations (FELASA).

Published in Russian in Biokhimiya, 2019, Vol. 84, No. 12, pp. 1854–1866.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM19–194, September 23, 2019.

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Andrianova, N.V., Zorova, L.D., Babenko, V.A. et al. Rapamycin Is Not Protective against Ischemic and Cisplatin-Induced Kidney Injury. Biochemistry Moscow 84, 1502–1512 (2019) doi:10.1134/S0006297919120095

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Keywords

  • rapamycin
  • acute kidney injury
  • ischemia
  • cisplatin
  • renal tubular cells
  • autophagy
  • nephroprotection