Related Experiment Video
Updated: Feb 20, 2026
A Doxorubicin-induced Cardiomyopathy Model in Adult Zebrafish
Published on: June 7, 2018
CRISPR/Cas9 Screens Implicate RARA and SPNS1 in Doxorubicin Cardiotoxicity
Chris McDermott-Roe1, Wenjian Lv1, Yeng Shao1
1Cardiovascular Institute, Department of Medicine, and Department of Genetics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Background:
Doxorubicin (DOX) causes cardiotoxicity and heart failure in a significant fraction of patients, but the molecular etiology is poorly understood.
Objectives:
We adopted a functional genomics-based approach to probe the genetic basis for DOX-induced cardiotoxicity in an exhaustive and agnostic manner.
Methods:
Genome-wide and targeted CRISPR/Cas9 screens were performed in immortalized cardiomyocytes and induced pluripotent stem cell-derived cardiomyocytes to identify genetic modifiers of DOX-induced cardiotoxicity.
Results:
Our first screen revealed that loss of the Retinoic Acid Receptor Alpha gene (RARA) increased DOX-induced cell death. Conversely, pharmacological activation of RARA protein with tamibarotene reduced DOX-induced toxicity. RNA-Seq analysis showed that whereas DOX broadly suppressed expression of metabolic and mitochondrial genes, tamibarotene mitigated this effect. In a second screen, we interrogated processes involved in DOX uptake, transport, and efflux. Loss of lysosome homeostasis, exemplified by SPNS lysolipid transporter 1 (SPNS1) deficiency, led to DOX hyperaccumulation, suppression of autophagy, increased DNA damage, and increased cell death. In contrast, ribosome loss-of-function and nutrient deprivation significantly reduced DOX accumulation and toxicity.
Conclusions:
Our study identified numerous drug-gene interactions that illuminate mechanisms underlying DOX-induced cardiotoxicity and provide a technical framework for future functional genomics screens to nominate therapeutic targets and genetic biomarkers.
Insights
Doxorubicin (DOX) causes heart damage, but this study used CRISPR screens to find genetic factors. Loss of RARA increased DOX toxicity, while tamibarotene reduced it, offering potential therapeutic targets.
Area of Science:
- Genomics
- Cardiology
- Drug Discovery
Background:
- Doxorubicin (DOX) is a chemotherapy agent known to induce cardiotoxicity and heart failure.
- The precise molecular mechanisms underlying DOX-induced cardiotoxicity remain incompletely understood.
Purpose of the Study:
- To employ a functional genomics approach to comprehensively identify genetic modifiers of DOX-induced cardiotoxicity.
- To uncover novel molecular pathways and potential therapeutic targets for mitigating DOX-induced heart damage.
Main Methods:
- Utilized genome-wide and targeted CRISPR/Cas9 screening in cardiomyocyte models.
- Performed RNA-Sequencing to analyze gene expression changes.
- Investigated genetic factors influencing DOX uptake, transport, and efflux.
Main Results:
- Loss of Retinoic Acid Receptor Alpha (RARA) exacerbated DOX-induced cell death; RARA activation with tamibarotene conferred protection.
- Tamibarotene treatment counteracted DOX-induced suppression of metabolic and mitochondrial gene expression.
- Disruption of lysosome homeostasis (e.g., SPNS1 deficiency) led to DOX hyperaccumulation and increased toxicity, while ribosome dysfunction and nutrient deprivation reduced DOX toxicity.
Conclusions:
- Identified key drug-gene interactions elucidating DOX cardiotoxicity mechanisms.
- Highlighted RARA and lysosome homeostasis as critical players in DOX response.
- Established a functional genomics framework for discovering therapeutic targets and biomarkers for DOX cardiotoxicity.
Related Concept Videos
01:28CRISPR/Cas9 Genome Editing
02:31Homologous Recombination
01:59CRISPR