TELOMERE LENGTH REGULATION IN WISTAR RATS UNDER SUBCHRONIC LOW-DOSE DOXORUBICIN EXPOSURE

Highlights

Doxorubicin is an anthracycline chemotherapeutic drug that stops proliferation, activates apoptosis, and has cumulative and dose–dependent cardiotoxicity. The cardiotoxicity of anthracyclines is based on damage to myocardial cells, accompanied by left ventricular dysfunction. Telomere dysfunction plays an important role in cellular aging processes that are associated with myocardial damage.

Aim. To evaluate the expression of genes encoding proteins involved in the regulation of the length of telomeric DNA regions of Wistar rat cardiomyocytes under subchronic low-dose doxorubicin exposure.

Methods. The study included two groups of male Wistar rats: an experimental group (10 rats, weekly injection of doxorubicin into the tail vein at a dosage of 2 mg/kg for 4 weeks) and a control group (10 rats, weekly injection of 0.9% NaCl into the tail vein for 4 weeks). The expression level of TERT and shelterin genes Trf1, Trf2, Rap1, Tin2, Tpp1 and Pot1 was determined using PCR.

Results. The experimental group was characterized by increased expression of the shelterin genes Trf1, Trf2, Tpp1 and Tin2 compared to the controls. The mRNA level of Pot1 and Rap1 was lower in the experimental group compared to the controls. The TERT gene was expressed at an extremely low level.

Conclusion. The study findings revealed that the lengthening of telomeric regions of DNA in Wistar rat cardiomyocytes in response to subchronic low-dose exposure to doxorubicin is due to changes in the expression profile of genes encoding proteins responsible for maintaining telomere homeostasis, and probably occurs due to the triggering of the mechanism of alternative lengthening of telomeres.

1. Li Y, Yan J, Yang P. The mechanism and therapeutic strategies in doxorubicin-induced cardiotoxicity: Role of programmed cell death. Cell Stress and Chaperones. 2024;29(5):666-680. doi: 10.1016/j.cstres.2024.09.001

2. Teplyakov AT, Shilov SN, Popova AA, et al. The role of polymorphic variants of NO-synthase, receptor endothelin-1 and NADPH-oxidase in the development of heart failure resulted from chemotherapy-induced cardiotoxicity. Complex Issues of Cardiovascular Diseases. 2018;7(3):33-43. doi: 10.17802/2306-1278-2018-7-3-33-43. (In Russian)

3. Zhang J., Jiang H., Zhang J., Bao G., Zhang G., Wang H., Wang X. Effectiveness and safety of pegylated liposomal doxorubicin versus epirubicin as neoadjuvant or adjuvant chemotherapy for breast cancer: a real-world study. BMC Cancer. 2021;21(1):1301. doi: 10.1186/s12885-021-09050-6

4. Sheibani M., Azizi Y., Shayan M., Nezamoleslami S., Eslami F., Farjoo M.H., Dehpour A.R. Doxorubicin-Induced Cardiotoxicity: An Overview on Pre-clinical Therapeutic Approaches. Cardiovascular Toxicology. 2022;22(4):292-310. doi: 10.1007/s12012-022-09721-1

5. Ivanová M., Dovinová I., Okruhlicová L., Tribulová N., Simončíková P., Barteková M., et al. Chronic cardiotoxicity of doxorubicin involves activation of myocardial and circulating matrix metalloproteinases in rats. Acta Pharmacol Sin. 2012; 33(4):459-69. doi:10.1038/aps.2011.194

6. Pod"yacheva E.YU., SHmakova T.V., Andreeva D.D., Toropov R.I., CHeburkin YU.V., Danil'chuk M.S. i dr. Profil' molekulyarnyh markerov fibroza u krys pri vozdejstvii razlichnyh doz doksorubicina. ZHurnal evolyucionnoj biohimii i fiziologii. 2023;59(2):121-130. doi: 10.31857/S0044452923020043 (In Russian)

7. Rossiello F., Jurk D., Passos J.F., d'Adda di Fagagna F. Telomere dysfunction in ageing and age-related diseases. Nature Cell Biology. 2022;24(2):135-147. doi: 10.1038/s41556-022-00842-x

8. Tsatsakis A., Oikonomopoulou T., Nikolouzakis T.K., Vakonaki E., Tzatzarakis M., Flamourakis M., Renieri E., Fragkiadaki P., et al. Role of telomere length in human carcinogenesis (Review). International Journal of Oncology. 2023;63(1):78. doi: 10.3892/ijo.2023.5526

9. Rubtsova M.P., Nikishin D.A., Vyssokikh M.Y., Koriagina M.S., Vasiliev A.V., Dontsova O.A. Telomere reprogramming and cellular metabolism: is there a link? International Journal of Molecular Sciences. 2024;25(19):10500. doi: 10.3390/ijms251910500

10. Asanov M.A., Poddubnyak A.O., Sinitskaya A.V., Khutornaya M.V., Khryachkova O.V. , Sinitsky M. Yu. Telomere length of Wistar rat cardiomyocytes exposed to doxorubicin: in vivo experimental study. Bulletin of Experimental Biology and Medicine. 2024;178(11):554-557. doi: 10.47056/0365-9615-2024-178-11-554-557. (In Russian)

11. Nair A., Morsy M.A., Jacob S. Dose translation between laboratory animals and human in preclinical and clinical phases of drug development. Drug Development Research. 2018;79(8):373-382. doi: 10.1002/ddr.21461

12. Asanov MA, Poddubnyak AO, Muhamadiyarov RA, Sinitskaya A.V., Khutornaya M.V., Sinitsky M.Yu. Comprehensive assessment of subchronic low-dose exposure to doxorubicin in the Wistar rat model. Siberian Journal of Clinical and Experimental Medicine. 2024;39(4):171-179. doi: 10.29001/2073-8552-2024-39-4-171-179. (In Russian).

13. Al Khafaji A.T., Barakat A.M., Shayyal A.J., Taan A.A., Aboqader Al-Aouadi R.F. Managing Doxorubicin Cardiotoxicity: Insights Into Molecular Mechanisms and Protective Strategies. Journal of Biochemical and Molecular Toxicology. 202539(2):e70155. doi: 10.1002/jbt.70155

14. Lyon A.R., López-Fernández T., Couch L.S., Asteggiano R., Aznar M.C., Bergler-Klein J., Boriani G., Cardinale D., et al. 2022 ESC Guidelines on cardio-oncology developed in collaboration with the European Hematology Association (EHA), the European Society for Therapeutic Radiology and Oncology (ESTRO) and the International Cardio-Oncology Society (IC-OS). European Heart Journal. 2022;43(41):4229-4361. doi: 10.1093/eurheartj/ehac244

15. Dempke W.C.M., Zielinski R., Winkler C., Silberman S., Reuther S., Priebe W. Anthracycline-induced cardiotoxicity - are we about to clear this hurdle? European Journal of Cancer. 2023;185:94-104. doi: 10.1016/j.ejca.2023.02.019

16. Gao F., Xu T., Zang F., Luo Y., Pan D. Cardiotoxicity of Anticancer Drugs: Molecular Mechanisms, Clinical Management and Innovative Treatment. Drug Design, Development and Therapy. 2024;18:4089-4116. doi: 10.2147/DDDT.S469331

17. Hansen M., Rubinsztein D.C., Walker D.W. Autophagy as a promoter of longevity: insights from model organisms. Nature Reviews Molecular Cell Biology. 2018; 19:579–593. doi: 10.1038/s41580-018-0033-y

18. Dixon S.J., Lemberg K.M., Lamprecht M.R., Skouta R., Zaitsev E.M., Gleason C.E., et al. Ferroptosis. An iron-dependent form of nonapoptotic cell death. Cell. 2012; 149(5):1060-1072. doi: 10.1016/j.cell.2012.03.042

19. Meng L., Lin H., Zhang J., Lin N., Sun Z., Gao F., et al. Doxorubicin induces cardiomyocyte pyroptosis via the TINCR-mediated posttranscriptional stabilization of NLR family pyrin domain containing 3. J Mol Cell Cardiol. 2019; 136:15-26. doi: 10.1016/j.yjmcc.2019.08.009

20. Wang T.H., Ma Y., Gao S., Zhang W.W., Han D., Cao F. Recent Advances in the Mechanisms of Cell Death and Dysfunction in Doxorubicin Cardiotoxicity. Reviews in Cardiovascular Medicine. 2023;24(11):336. doi: 10.31083/j.rcm2411336

21. Lin J., Epel E. Stress and telomere shortening: Insights from cellular mechanisms. Ageing Research Reviews. 2022;73:101507. doi: 10.1016/j.arr.2021.101507

22. Vaiserman A., Krasnienkov D. Telomere Length as a Marker of Biological Age: State-of-the-Art, Open Issues, and Future Perspectives. Frontiers in Genetics. 202121;11:630186. doi: 10.3389/fgene.2020.630186

23. Liu M., Zhang Y., Jian Y., et al. The regulations of telomerase reverse transcriptase (TERT) in cancer. Cell Death & Disease. 2024;15(1):90. doi: 10.1038/s41419-024-06454-7

24. Chatterjee S., Leach-Mehrwald M., Huang C.K., Xiao K., Fuchs M., Otto M., Lu D., Dang V., et al. Telomerase is essential for cardiac differentiation and sustained metabolism of human cardiomyocytes. Cellular and Molecular Life Sciences. 2024;81(1):196. doi: 10.1007/s00018-024-05239-7

25. Wu Y., Zhou L., Liu H., Duan R., Zhou H., Zhang F., He X., Lu D., et al. LRP6 downregulation promotes cardiomyocyte proliferation and heart regeneration. Cell Research. 2021;31(4):450-462. doi: 10.1038/s41422-020-00411-7

26. Lim C.J., Cech T.R. Shaping human telomeres: From shelterin and CST complexes to telomeric chromatin organization. Nature Reviews Molecular Cell Biology. 2021;22(4):283-298. doi: 10.1038/s41580-021-00328-y

27. Mir S.M., Samavarchi Tehrani S., Goodarzi G., Jamalpoor Z., Asadi J., Khelghati N., Qujeq D,. Maniati M. Shelterin Complex at Telomeres: Implications in Ageing. Clinical Interventions in Aging. 2020;15:827-839. doi: 10.2147/CIA.S256425

28. Kallingal A., Krzemieniecki R., Maciejewska N., Brankiewicz-Kopcińska W., Baginski M. TRF1 and TRF2: pioneering targets in telomere-based cancer therapy. Journal of Cancer Research and Clinical Oncology. 2024;150(7):353. doi: 10.1007/s00432-024-05867-3

29. Deregowska A., Lewinska A., Warzybok A., Stoklosa T., Wnuk M. Telomere loss is accompanied by decreased pool of shelterin proteins TRF2 and RAP1, elevated levels of TERRA and enhanced glycolysis in imatinib-resistant CML cells. Toxicology in Vitro. 2023;90:105608. doi: 10.1016/j.tiv.2023.105608

30. Deregowska A., Wnuk M. RAP1/TERF2IP-A Multifunctional Player in Cancer Development. Cancers. 2021;13(23):5970. doi: 10.3390/cancers13235970

31. Ueno M. Exploring Genetic Interactions with Telomere Protection Gene POT1 in Fission Yeast. Biomolecules. 2023;13(2):370. doi: 10.3390/biom13020370

32. Sohn E.J., Goralsky J.A., Shay J.W., Min J. The Molecular Mechanisms and Therapeutic Prospects of Alternative Lengthening of Telomeres (ALT). Cancers (Basel). 2023;15(7):1945. doi: 10.3390/cancers15071945

33. Cesare A.J., Kaul Z., Cohen S.B., Napier C.E., Pickett H.A., Neumann A.A., Reddel R.R. Spontaneous occurrence of telomeric DNA damage response in the absence of chromosome fusions. Nature Structural & Molecular Biology. 2009;16(12):1244-1251. doi: 10.1038/nsmb.1725