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

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, ho...

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Autores principales:	Valerie Sapp, Aitor Aguirre, Gayatri Mainkar, Jeffrey Ding, Eric Adler, Ronglih Liao, Sonia Sharma, Mohit Jain
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3e7b6bbe20104376830c9ac969a7c0df
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spelling	oai:doaj.org-article:3e7b6bbe20104376830c9ac969a7c0df2021-12-02T15:39:59ZGenome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity10.1038/s41598-021-92988-12045-2322https://doaj.org/article/3e7b6bbe20104376830c9ac969a7c0df2021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92988-1https://doaj.org/toc/2045-2322Abstract 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.Valerie SappAitor AguirreGayatri MainkarJeffrey DingEric AdlerRonglih LiaoSonia SharmaMohit JainNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Valerie Sapp Aitor Aguirre Gayatri Mainkar Jeffrey Ding Eric Adler Ronglih Liao Sonia Sharma Mohit Jain Genome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity
description	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.
format	article
author	Valerie Sapp Aitor Aguirre Gayatri Mainkar Jeffrey Ding Eric Adler Ronglih Liao Sonia Sharma Mohit Jain
author_facet	Valerie Sapp Aitor Aguirre Gayatri Mainkar Jeffrey Ding Eric Adler Ronglih Liao Sonia Sharma Mohit Jain
author_sort	Valerie Sapp
title	Genome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity
title_short	Genome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity
title_full	Genome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity
title_fullStr	Genome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity
title_full_unstemmed	Genome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity
title_sort	genome-wide crispr/cas9 screening in human ips derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/3e7b6bbe20104376830c9ac969a7c0df
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Genome-wide CRISPR/Cas9 screening in human iPS derived cardiomyocytes uncovers novel mediators of doxorubicin cardiotoxicity

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