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Cited by 40CRISPR and Genetic EngineeringChemotherapy-induced cardiotoxicity and mitigationPluripotent Stem Cells Research

Genome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity

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Sapp, Valerie, Aguirre, Aitor, Mainkar, Gayatri, Ding, Jeffrey, Adler, Eric, Liao, Ronglih, Sharma, Sonia, Jain, Mohit·2021·Scientific Reports
DOI 10.1038/s41598-021-92988-1PMC PMC8260754
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Abstract

Human induced pluripotent stem (iPS) cell technologies coupled with genetic engineering now facilitate the study of the molecular underpinnings of disease in relevant human cell types. Application of CRISPR/Cas9-based approaches for genome-scale functional screening in iPS-derived cells, however, has been limited by technical constraints, including inefficient transduction in pooled format, loss of library representation, and poor cellular differentiation. Herein, we present optimized approaches for whole-genome CRISPR/Cas9 based screening in human iPS derived cardiomyocytes with near genome-wide representation at both the iPS and differentiated cell stages. As proof-of-concept, we perform a screen to investigate mechanisms underlying doxorubicin mediated cell death in iPS derived cardiomyocytes. We identified two poorly characterized, human-specific transporters (SLCO1A2, SLCO1B3) whose loss of function protects against doxorubicin-cardiotoxicity, but does not affect cell death in cancer cells. This study provides a technical framework for genome-wide functional screening in iPS derived cells and identifies new targets to mitigate doxorubicin-cardiotoxicity in humans.

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Cited by

Sorted by citations

Authors
Sapp, V., Aguirre, A., Mainkar, G., Ding, J., Adler, E., Liao, R., Sharma, S., & Jain, M.
Affiliations
  • University of California San Diego (US)
  • Stanford University (US)
  • La Jolla Institute for Immunology (US)
Corresponding author
Jain, Mohit
ORCID
Venue
Scientific Reports
Publisher
Springer Science and Business Media LLC
Published
2021-07-06
Volume
11
Issue
1
Pages
13866-13866
ISSN
2045-2322 (ISSN-L)
DOI
10.1038/s41598-021-92988-1
PMC
PMC8260754
OpenAlex
W3179677353
Cited by
40
References
6
Citations by year
2026: 9 · 2025: 6 · 2024: 11 · 2023: 6 · 2022: 6 · 2021: 2
FWCI
2.90
Type
journal article
Language
en
Open access
Licence
Locations
  • Scientific Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Institutes of Health (T32GM007752, K01HL135464, K23HL107755, R01HL128135, R01CA199376, R03HL133720)
  • University of California, San Diego (Research Excellence Scholarships)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Chemotherapy-induced cardiotoxicity and mitigation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
Keywords
doxorubicin cardiotoxicity, SLCO1A2, SLCO1B3, drug transporters, cardiomyocyte death, iPSC differentiation
MeSH terms
RNA, Guide, CRISPR-Cas Systems, Doxorubicin, Humans, Phenotype, Genome, Human, Lentivirus, Myocytes, Cardiac, Cell Line, Tumor, Induced Pluripotent Stem Cells, CRISPR-Cas Systems, Cardiotoxicity

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