SiRNA-mediated knockdown of TOP2B protects hiPSC-derived cardiomyocytes from doxorubicin-induced toxicity

IF 5.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Life sciences Pub Date : 2025-03-28 DOI:10.1016/j.lfs.2025.123595

Neha Saroj , Pankaj Singh Dholaniya , Syed Baseeruddin Alvi , Divya Sridharan , Navisha Soni , Syed Abdullah Ashraf , Ayza Choudhry , Yusuf Ali Ashraf , Sarah Kathleen Mikula , Dinender Kumar Singla , Mahmood Khan

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引用次数: 0

Abstract

Aims

Doxorubicin (Dox) is a potent chemotherapeutic agent, but its use is limited by cardiotoxicity, primarily due to the disruption of Topoisomerase-2 beta (TOP2B) activity. Dexrazoxane (Dex), an FDA-approved cardioprotective drug, alleviates Dox-induced toxicity but lacks heart-specific targeting. This study investigates siRNA-mediated TOP2B knockdown as a more targeted strategy to protect cardiomyocytes from Dox-induced damage.

Materials and methods

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were treated with siRNA to knock down TOP2B and were then exposed to Dox. We compared the cardioprotective effects of siRNA-mediated knockdown to Dex treatment using cell viability, cell toxicity assay and electrophysiological evaluation was performed using a multielectrode array (MEA).

Key findings

Our results demonstrate that TOP2B silencing significantly decreases apoptosis and improved cell viability, as compared to the Dex treatment. Additionally, electrophysiological assays using a Multielectrode Array (MEA) demonstrated enhanced contractility and conductivity in siRNA-treated hiPSC-CMs. Furthermore, transmission electron microscopy (TEM) data revealed that TOP2B knockdown preserves mitochondrial morphology and sarcomere structure, compared to Dox and Dex-treated groups.

Significance

These findings suggest that siRNA-mediated TOP2B inhibition could provide a safer, more specific approach to mitigate Dox-induced cardiotoxicity.

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sirna介导的TOP2B敲低可保护hipsc来源的心肌细胞免受阿霉素诱导的毒性

目的多柔比星（Dox）是一种有效的化疗药物，但其使用受到心脏毒性的限制，主要是由于拓扑异构酶-2 β (TOP2B)活性的破坏。Dexrazoxane （Dex）是一种fda批准的心脏保护药物，可减轻dox诱导的毒性，但缺乏心脏特异性靶向。本研究探讨了sirna介导的TOP2B敲低作为一种更有针对性的策略来保护心肌细胞免受dox诱导的损伤。材料和方法用siRNA处理人诱导多能干细胞来源的心肌细胞（hiPSC-CMs），敲低TOP2B，然后暴露于Dox。我们通过细胞活力、细胞毒性试验和使用多电极阵列（MEA）进行电生理评估，比较了sirna介导的敲低和Dex治疗的心脏保护作用。结果表明，与Dex处理相比，TOP2B沉默可显著减少细胞凋亡，提高细胞活力。此外，使用多电极阵列（MEA）的电生理分析表明，sirna处理的hiPSC-CMs具有增强的收缩性和导电性。此外，透射电镜（TEM）数据显示，与Dox和dex处理组相比，TOP2B敲除保留了线粒体形态和肌节结构。这些发现表明，sirna介导的TOP2B抑制可以提供一种更安全、更特异性的方法来减轻dox诱导的心脏毒性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Life sciences 医学-药学

CiteScore

12.20

自引率

1.60%

发文量

841

审稿时长

6 months

期刊介绍： Life Sciences is an international journal publishing articles that emphasize the molecular, cellular, and functional basis of therapy. The journal emphasizes the understanding of mechanism that is relevant to all aspects of human disease and translation to patients. All articles are rigorously reviewed. The Journal favors publication of full-length papers where modern scientific technologies are used to explain molecular, cellular and physiological mechanisms. Articles that merely report observations are rarely accepted. Recommendations from the Declaration of Helsinki or NIH guidelines for care and use of laboratory animals must be adhered to. Articles should be written at a level accessible to readers who are non-specialists in the topic of the article themselves, but who are interested in the research. The Journal welcomes reviews on topics of wide interest to investigators in the life sciences. We particularly encourage submission of brief, focused reviews containing high-quality artwork and require the use of mechanistic summary diagrams.