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Updated: Jun 27, 2026
An In Vitro 3D Model and Computational Pipeline to Quantify the Vasculogenic Potential of iPSC-Derived Endothelial Progenitors
Published on: May 13, 2019
Endothelial progenitor cell susceptibility to DNA damaging and DDR-modulating compounds determines endothelial
Sina Federmann1, Michelle Westerhoff2, Andreas S Reichert2
1Institute of Toxicology, Medical Faculty and University Hospital, Heinrich-Heine-University Duesseldorf, Moorenstrasse 5, 40225, Duesseldorf, Germany.
Insights
Anticancer drug doxorubicin (Dox) damages cardiac cells, impacting regeneration. Protecting endothelial progenitor cells (EPCs) from Dox damage is crucial for maintaining heart health and preventing cardiotoxicity.
Area of Science:
- Cardiovascular Biology
- Cancer Therapeutics
- Cellular Regeneration
Background:
- Doxorubicin (Dox) is a vital anticancer drug, but its use is limited by cardiotoxicity.
- The role of specific cardiac cell types, particularly endothelial progenitor cells (EPCs), in Dox-induced cardiotoxicity is not fully understood.
- Understanding cellular responses to Dox is critical for mitigating its adverse effects.
Purpose of the Study:
- To investigate the differential responses of murine embryonic stem cells (mESC), endothelial progenitor cells (EC d4), and differentiated endothelial-like cells (EC d6) to Dox and DNA damage response inhibitors.
- To determine if Dox treatment affects the differentiation and function of EPCs and their progeny.
- To explore potential strategies for protecting EPCs and preserving endothelial function during cancer therapy.
Main Methods:
- Comparative analysis of mESC, EC d4, and EC d6 responses to Dox, RAD51 inhibitor (B02), and HDAC inhibitor (entinostat).
- Assessment of DNA damage (SSB, DSB), cell proliferation (EdU incorporation), and replication fork progression.
- Evaluation of functional parameters in differentiated progeny, including mitochondrial homeostasis, endothelial barrier integrity (ZO-1, VE-cadherin), cytokine response, LDL uptake, and senescence.
Main Results:
- Endothelial progenitor cells (EC d4) showed higher sensitivity to Dox compared to mESC and EC d6.
- Differential DNA damage profiles were observed: mESC had more single-strand breaks (SSB), while EC d6 had more double-strand breaks (DSB).
- Dox-treated EC d4 differentiated into EC d6, but these progeny exhibited impaired mitochondrial function, compromised endothelial barrier, altered cytokine response, reduced LDL uptake, and increased senescence.
Conclusions:
- Both overlapping and distinct cellular responses to Dox and DNA repair inhibitors were identified across cell types.
- Dox treatment of EPCs leads to functional deficits in their differentiated endothelial progeny.
- Targeted protection of EPCs from Dox-induced damage is recommended to maintain endothelial health and reduce long-term cardiotoxicity risk.
Abstract:
The clinical use of the anticancer drug doxorubicin (Dox) is limited by irreversible cardiotoxicity. The pathophysiological relevance of different cardiac cell types, including endothelial progenitor cells (EPC), in this process is unclear. Since progenitor cells are particularly relevant for tissue regeneration, we hypothesize that residual damage resulting from Dox-based therapeutic regimen may influence their endothelial differentiation accuracy. Therefore, we comparatively investigated the response of murine embryonic stem cells (mESC), endothelial progenitor cells (EC d4) and terminally differentiated endothelial-like cells (EC d6) following exposure to Dox and selected pharmacological inhibitors of DNA repair/DNA damage response (DDR) (RAD51i B02; HDACi entinostat (EST)). We show that EC d4 exhibit enhanced Dox sensitivity as compared to mESC and EC d6. EdU incorporation and replication fork progression analyses revealed pronounced agent-specific differences between mESC, EC d4 and EC d6. Furthermore, DNA damage formation varied in a drug-dependent manner, with mESC showing enhanced residual levels of DNA single-strand breaks (SSB) as compared to EC d4 and EC d6 while EC d6 revealed highest levels of DNA double-strand breaks (DSB). Dox treatment of EC d4 did not prevent their further differentiation into EC d6. However, it caused several functional impairments in the surviving EC d6 progeny, including defects in mitochondrial homeostasis, endothelial barrier function related to cell-cell adhesion factors (ZO1, VE-cadherin), cytokine response and low-density lipoprotein (LDL) uptake. This is accompanied by increased senescence. Summarizing, we demonstrate both overlapping and agent-specific responses of mESC, EC d4 and EC d6 to Dox and DNA repair/DDR inhibitors. Notably, drug treatment of EPC (EC d4) causes multiple dysfunctions in differentiated EC d6. Hence, pharmacological measures aiming to specifically protect EPC from Dox-induced damage are suggested to foster the maintenance of healthy endothelial functionality during regeneration, thereby lowering the risk of detrimental late cardiotoxicity resulting from Dox-based anticancer regimen.
