Research Paper|Volume 15, Issue 1|pp 164—178

Doxorubicin induced ROS-dependent HIF1α activation mediates blockage of IGF1R survival signaling by IGFBP3 promotes cardiac apoptosis

Su-Ying Wen^1,^2,³, Ayaz Ali⁴, I-Chieh Huang⁴, Jian-Sheng Liu^4,⁵, Po-Yuan Chen⁴, Vijaya Padma Viswanadha⁶, Chih-Yang Huang^7,^8,^9,^10,¹¹, Wei-Wen Kuo^4,¹²

¹Department of Dermatology, Taipei City Hospital, Renai Branch, Taipei 11260, Taiwan
²Department of Cosmetic Applications and Management, Mackay Junior College of Medicine, Nursing and Management, Taipei 112, Taiwan
³Department of Health Care Management, National Taipei University of Nursing and Health Sciences, Taipei, Taiwan
⁴Department of Biological Science and Technology, China Medical University, Taichung 404, Taiwan
⁵China Medical University Beigang Hospital Thoracic Department, Yunlin 651, Taiwan
⁶Department of Biotechnology, Bharathiar University, Coimbatore 641046, India
⁷Center of General Education, Buddhist Tzu Chi Medical Foundation, Tzu Chi University of Science and Technology, Hualien 970, Taiwan
⁸Department of Medical Research, China Medical University Hospital, China Medical University, Taichung 404, Taiwan
⁹Department of Medical Laboratory Science and Biotechnology, Asia University, Taichung 413, Taiwan
¹⁰Graduate Institute of Biomedical Sciences, China Medical University, Taichung 404, Taiwan
¹¹Cardiovascular and Mitochondrial Related Disease Research Center, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Hualien 970, Taiwan
¹²Ph.D. Program for Biotechnology Industry, China Medical University, Taichung 406, Taiwan

* Equal contribution

Received: November 11, 2022Accepted: December 5, 2022Published: January 3, 2023

https://doi.org/10.18632/aging.204466

Copyright: © 2023 Wen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Doxorubicin (Dox) causes the generation of intracellular reactive oxygen species (ROS) and inactivates insulin-like growth factor 1 (IGF1) signaling, leading to cardiomyocyte apoptosis. IGF-binding protein 3 (IGFBP3) is the most abundant circulating IGF1 carrier protein with high affinity, which has been reported to mediate ROS-induced apoptosis. Hypoxia-inducible factor 1α (HIF1A), an upstream protein of IGFBP3 is regulated by prolyl hydroxylase domain (PHD) through hydroxylation. In this study, we investigated the role of IGFBP3, HIF1A, and PHD in Dox-induced cardiac apoptosis.Cells challenged with 1 μM Dox for 24 h increased ROS generation, augmented intracellular and secreted IGFBP3 levels, and reduced IGF1 signaling. Further, we showed that Dox enhanced the extracellular association of IGF1 with IGFBP3. Moreover, echocardiography parameters, especially ejection fraction (EF) and fractional shortening (FS) were significantly reduced in ventricle tissue of Dox challenged rats. Notably, siRNA approach against IGFBP3 or an anti-IGFBP3 antibody rescued Dox-induced cardiac apoptosis, mitochondrial ROS, and the decrease in the IGF1 signaling activity. Furthermore, silencing HIF1A either using siRNA or inhibitor downregulated intracellular IGFBP3, rescued apoptosis, mitochondrial generation, and reduction in IGF1 signaling. Finally, western blot data revealed that ROS scavenger reversed Dox-induced cardiac apoptosis, increased levels of HIF1A and secreted IGFBP3, and decreased IGF1 survival signaling and PHD expression.

These findings suggest that Dox-induced ROS generation suppressed PHD, which might stabilize nuclear HIF1A protein, leading to increased IGFBP3 expression and secretion. This in turn results in enhanced extracellular association of the latter with IGF1 and blocks IGF1 pro-survival signaling and may result in inducing cardiac apoptosis.