Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

Sridhar Selvaraj; Ricardo Mondragon-Gonzalez; Bin Xu; Alessandro Magli; Hyunkee Kim; Jeanne Lainé; James Kiley; Holly McKee; Fabrizio Rinaldi; Joy Aho; Nacira Tabti; Wei Shen; Rita Cr Perlingeiro

doi:10.7554/eLife.47970

Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

Sridhar Selvaraj, Ricardo Mondragon-Gonzalez, Bin Xu, Alessandro Magli, Hyunkee Kim, Jeanne Lainé, James Kiley, Holly McKee, Fabrizio Rinaldi, Joy Aho, Nacira Tabti, Wei Shen, Rita Cr Perlingeiro

Research output: Contribution to journal › Article › peer-review

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Abstract

Targeted differentiation of pluripotent stem (PS) cells into myotubes enables in vitro disease modeling of skeletal muscle diseases. Although various protocols achieve myogenic differentiation in vitro, resulting myotubes typically display an embryonic identity. This is a major hurdle for accurately recapitulating disease phenotypes in vitro, as disease commonly manifests at later stages of development. To address this problem, we identified four factors from a small molecule screen whose combinatorial treatment resulted in myotubes with enhanced maturation, as shown by the expression profile of myosin heavy chain isoforms, as well as the upregulation of genes related with muscle contractile function. These molecular changes were confirmed by global chromatin accessibility and transcriptome studies. Importantly, we also observed this maturation in three-dimensional muscle constructs, which displayed improved in vitro contractile force generation in response to electrical stimulus. Thus, we established a model for in vitro muscle maturation from PS cells.

Original language	English (US)
Article number	e47970
Journal	eLife
Volume	8
DOIs	https://doi.org/10.7554/eLife.47970
State	Published - Nov 2019

Bibliographical note

Funding Information:
This project was supported by funds from the NIH, grants R01 AR055299 and AR071439 (RCRP) and the NSF CAREER DMR-1151529 (WS). RM-G was funded by CONACyT-Mexico (#394378). We also thank the generous support from ADVault, Inc and MyDirectives.com (RCRP). The cytogenetic analyses were performed in the Cytogenomics Shared Resource at the University of Minnesota with support from the comprehensive Masonic Cancer Center NIH Grant #P30 CA077598-09. The monoclonal antibodies to MyHC, Titin, MyHC-neo and MYH1/2 were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa. We thank Prof. Robert T Tranquillo, Dr. Jeremy Schaefer, and Dr. Lauren Black for the use of the contractile force measurement apparatus and for facilitating data analysis.

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© 2019, eLife Sciences Publications Ltd. All rights reserved.

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title = "Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes",

abstract = "Targeted differentiation of pluripotent stem (PS) cells into myotubes enables in vitro disease modeling of skeletal muscle diseases. Although various protocols achieve myogenic differentiation in vitro, resulting myotubes typically display an embryonic identity. This is a major hurdle for accurately recapitulating disease phenotypes in vitro, as disease commonly manifests at later stages of development. To address this problem, we identified four factors from a small molecule screen whose combinatorial treatment resulted in myotubes with enhanced maturation, as shown by the expression profile of myosin heavy chain isoforms, as well as the upregulation of genes related with muscle contractile function. These molecular changes were confirmed by global chromatin accessibility and transcriptome studies. Importantly, we also observed this maturation in three-dimensional muscle constructs, which displayed improved in vitro contractile force generation in response to electrical stimulus. Thus, we established a model for in vitro muscle maturation from PS cells.",

author = "Sridhar Selvaraj and Ricardo Mondragon-Gonzalez and Bin Xu and Alessandro Magli and Hyunkee Kim and Jeanne Lain{\'e} and James Kiley and Holly McKee and Fabrizio Rinaldi and Joy Aho and Nacira Tabti and Wei Shen and Perlingeiro, {Rita Cr}",

note = "Funding Information: This project was supported by funds from the NIH, grants R01 AR055299 and AR071439 (RCRP) and the NSF CAREER DMR-1151529 (WS). RM-G was funded by CONACyT-Mexico (#394378). We also thank the generous support from ADVault, Inc and MyDirectives.com (RCRP). The cytogenetic analyses were performed in the Cytogenomics Shared Resource at the University of Minnesota with support from the comprehensive Masonic Cancer Center NIH Grant #P30 CA077598-09. The monoclonal antibodies to MyHC, Titin, MyHC-neo and MYH1/2 were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa. We thank Prof. Robert T Tranquillo, Dr. Jeremy Schaefer, and Dr. Lauren Black for the use of the contractile force measurement apparatus and for facilitating data analysis. Publisher Copyright: {\textcopyright} 2019, eLife Sciences Publications Ltd. All rights reserved.",

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AU - Mondragon-Gonzalez, Ricardo

AU - Xu, Bin

AU - Magli, Alessandro

AU - Kim, Hyunkee

AU - Lainé, Jeanne

AU - Kiley, James

AU - McKee, Holly

AU - Rinaldi, Fabrizio

AU - Aho, Joy

AU - Tabti, Nacira

AU - Shen, Wei

AU - Perlingeiro, Rita Cr

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N2 - Targeted differentiation of pluripotent stem (PS) cells into myotubes enables in vitro disease modeling of skeletal muscle diseases. Although various protocols achieve myogenic differentiation in vitro, resulting myotubes typically display an embryonic identity. This is a major hurdle for accurately recapitulating disease phenotypes in vitro, as disease commonly manifests at later stages of development. To address this problem, we identified four factors from a small molecule screen whose combinatorial treatment resulted in myotubes with enhanced maturation, as shown by the expression profile of myosin heavy chain isoforms, as well as the upregulation of genes related with muscle contractile function. These molecular changes were confirmed by global chromatin accessibility and transcriptome studies. Importantly, we also observed this maturation in three-dimensional muscle constructs, which displayed improved in vitro contractile force generation in response to electrical stimulus. Thus, we established a model for in vitro muscle maturation from PS cells.

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Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

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