Evaluation of Senescence and Its Prevention in Doxorubicin-Induced Cardiotoxicity Using Dynamic Engineered Heart Tissues

Annet N Linders; Itamar B Dias; Ekaterina S Ovchinnikova; Mathilde C S C Vermeer; Martijn F Hoes; George Markousis Mavrogenis; Frederik E Deiman; Karla F Arevalo Gomez; Jacqueline M Bliley; Jamil Nehme; Aryan Vink; Jourik Gietema; Rudolf A de Boer; Daan Westenbrink; Herman H W Sillje; Denise Hilfiker-Kleiner; Linda W van Laake; Adam W Feinberg; Marco Demaria; Nils Bomer; Peter van der Meer

doi:10.1016/j.jaccao.2023.03.012

Evaluation of Senescence and Its Prevention in Doxorubicin-Induced Cardiotoxicity Using Dynamic Engineered Heart Tissues

JACC CardioOncol. 2023 May 16;5(3):298-315. doi: 10.1016/j.jaccao.2023.03.012. eCollection 2023 Jun.

Authors

Annet N Linders¹, Itamar B Dias¹, Ekaterina S Ovchinnikova¹, Mathilde C S C Vermeer¹, Martijn F Hoes^{2

3

4}, George Markousis Mavrogenis¹, Frederik E Deiman¹, Karla F Arevalo Gomez¹, Jacqueline M Bliley⁵, Jamil Nehme⁶, Aryan Vink⁷, Jourik Gietema⁸, Rudolf A de Boer¹, Daan Westenbrink¹, Herman H W Sillje¹, Denise Hilfiker-Kleiner⁹, Linda W van Laake¹⁰, Adam W Feinberg⁵, Marco Demaria⁶, Nils Bomer¹, Peter van der Meer¹

Affiliations

¹ Department of Cardiology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
² Department of Clinical Genetics, Maastricht University Medical Center+, Maastricht, the Netherlands.
³ CARIM School for Cardiovascular Diseases, Maastricht, the Netherlands.
⁴ Department of Cardiology, Faculty of Health, Medicine and Life Sciences, Maastricht University, Maastricht, the Netherlands.
⁵ Regenerative Biomaterials and Therapeutics Group, Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania.
⁶ European Research Institute for the Biology of Ageing, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
⁷ Department of Pathology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands.
⁸ Department of Medical Oncology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
⁹ Institute of Cardiovascular Complications in Pregnancy and in Oncologic Therapies, Philipps-Universität Marburg, Marburg, Germany.
¹⁰ Division of Heart and Lungs and Regenerative Medicine Center, University Medical Center Utrecht, Utrecht, the Netherlands.

Abstract

Background: Doxorubicin is an essential cancer treatment, but its usefulness is hampered by the occurrence of cardiotoxicity. Nevertheless, the pathophysiology underlying doxorubicin-induced cardiotoxicity and the respective molecular mechanisms are poorly understood. Recent studies have suggested involvement of cellular senescence.

Objectives: The aims of this study were to establish whether senescence is present in patients with doxorubicin-induced cardiotoxicity and to investigate if this could be used as a potential treatment target.

Methods: Biopsies from the left ventricles of patients with severe doxorubicin-induced cardiotoxicity were compared with control samples. Additionally, senescence-associated mechanisms were characterized in 3-dimensional dynamic engineered heart tissues (dyn-EHTs) and human pluripotent stem cell-derived cardiomyocytes. These were exposed to multiple, clinically relevant doses of doxorubicin to recapitulate patient treatment regimens. To prevent senescence, dyn-EHTs were cotreated with the senomorphic drugs 5-aminoimidazole-4-carboxamide ribonucleotide and resveratrol.

Results: Senescence-related markers were significantly up-regulated in the left ventricles of patients with doxorubicin-induced cardiotoxicity. Treatment of dyn-EHTs resulted in up-regulation of similar senescence markers as seen in the patients, accompanied by tissue dilatation, decreased force generation, and increased troponin release. Treatment with senomorphic drugs led to decreased expression of senescence-associated markers, but this was not accompanied by improved function.

Conclusions: Senescence was observed in the hearts of patients with severe doxorubicin-induced cardiotoxicity, and this phenotype can be modeled in vitro by exposing dyn-EHTs to repeated clinically relevant doses of doxorubicin. The senomorphic drugs 5-aminoimidazole-4-carboxamide ribonucleotide and resveratrol prevent senescence but do not result in functional improvements. These findings suggest that preventing senescence by using a senomorphic during doxorubicin administration might not prevent cardiotoxicity.

Keywords: cardiotoxicity; doxorubicin; hPSC-derived cardiomyocytes; senescence; senomorphic drugs.