Cargando…

Defining Reprogramming Checkpoints from Single-Cell Analyses of Induced Pluripotency

Elucidating the mechanism of reprogramming is confounded by heterogeneity due to the low efficiency and differential kinetics of obtaining induced pluripotent stem cells (iPSCs) from somatic cells. Therefore, we increased the efficiency with a combination of epigenomic modifiers and signaling molecu...

Descripción completa

Detalles Bibliográficos
Autores principales: Tran, Khoa A., Pietrzak, Stefan J., Zaidan, Nur Zafirah, Siahpirani, Alireza Fotuhi, McCalla, Sunnie Grace, Zhou, Amber S., Iyer, Gopal, Roy, Sushmita, Sridharan, Rupa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6555151/
https://www.ncbi.nlm.nih.gov/pubmed/31067459
http://dx.doi.org/10.1016/j.celrep.2019.04.056
_version_ 1783425100540805120
author Tran, Khoa A.
Pietrzak, Stefan J.
Zaidan, Nur Zafirah
Siahpirani, Alireza Fotuhi
McCalla, Sunnie Grace
Zhou, Amber S.
Iyer, Gopal
Roy, Sushmita
Sridharan, Rupa
author_facet Tran, Khoa A.
Pietrzak, Stefan J.
Zaidan, Nur Zafirah
Siahpirani, Alireza Fotuhi
McCalla, Sunnie Grace
Zhou, Amber S.
Iyer, Gopal
Roy, Sushmita
Sridharan, Rupa
author_sort Tran, Khoa A.
collection PubMed
description Elucidating the mechanism of reprogramming is confounded by heterogeneity due to the low efficiency and differential kinetics of obtaining induced pluripotent stem cells (iPSCs) from somatic cells. Therefore, we increased the efficiency with a combination of epigenomic modifiers and signaling molecules and profiled the transcriptomes of individual reprogramming cells. Contrary to the established temporal order, somatic gene inactivation and upregulation of cell cycle, epithelial, and early pluripotency genes can be triggered independently such that any combination of these events can occur in single cells. Sustained co-expression of Epcam, Nanog, and Sox2 with other genes is required to progress toward iPSCs. Ehf, Phlda2, and translation initiation factor Eif4a1 play functional roles in robust iPSC generation. Using regulatory network analysis, we identify a critical role for signaling inhibition by 2i in repressing somatic expression and synergy between the epigenomic modifiers ascorbic acid and a Dot1L inhibitor for pluripotency gene activation.
format Online
Article
Text
id pubmed-6555151
institution National Center for Biotechnology Information
language English
publishDate 2019
record_format MEDLINE/PubMed
spelling pubmed-65551512019-06-07 Defining Reprogramming Checkpoints from Single-Cell Analyses of Induced Pluripotency Tran, Khoa A. Pietrzak, Stefan J. Zaidan, Nur Zafirah Siahpirani, Alireza Fotuhi McCalla, Sunnie Grace Zhou, Amber S. Iyer, Gopal Roy, Sushmita Sridharan, Rupa Cell Rep Article Elucidating the mechanism of reprogramming is confounded by heterogeneity due to the low efficiency and differential kinetics of obtaining induced pluripotent stem cells (iPSCs) from somatic cells. Therefore, we increased the efficiency with a combination of epigenomic modifiers and signaling molecules and profiled the transcriptomes of individual reprogramming cells. Contrary to the established temporal order, somatic gene inactivation and upregulation of cell cycle, epithelial, and early pluripotency genes can be triggered independently such that any combination of these events can occur in single cells. Sustained co-expression of Epcam, Nanog, and Sox2 with other genes is required to progress toward iPSCs. Ehf, Phlda2, and translation initiation factor Eif4a1 play functional roles in robust iPSC generation. Using regulatory network analysis, we identify a critical role for signaling inhibition by 2i in repressing somatic expression and synergy between the epigenomic modifiers ascorbic acid and a Dot1L inhibitor for pluripotency gene activation. 2019-05-07 /pmc/articles/PMC6555151/ /pubmed/31067459 http://dx.doi.org/10.1016/j.celrep.2019.04.056 Text en This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Tran, Khoa A.
Pietrzak, Stefan J.
Zaidan, Nur Zafirah
Siahpirani, Alireza Fotuhi
McCalla, Sunnie Grace
Zhou, Amber S.
Iyer, Gopal
Roy, Sushmita
Sridharan, Rupa
Defining Reprogramming Checkpoints from Single-Cell Analyses of Induced Pluripotency
title Defining Reprogramming Checkpoints from Single-Cell Analyses of Induced Pluripotency
title_full Defining Reprogramming Checkpoints from Single-Cell Analyses of Induced Pluripotency
title_fullStr Defining Reprogramming Checkpoints from Single-Cell Analyses of Induced Pluripotency
title_full_unstemmed Defining Reprogramming Checkpoints from Single-Cell Analyses of Induced Pluripotency
title_short Defining Reprogramming Checkpoints from Single-Cell Analyses of Induced Pluripotency
title_sort defining reprogramming checkpoints from single-cell analyses of induced pluripotency
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6555151/
https://www.ncbi.nlm.nih.gov/pubmed/31067459
http://dx.doi.org/10.1016/j.celrep.2019.04.056
work_keys_str_mv AT trankhoaa definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency
AT pietrzakstefanj definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency
AT zaidannurzafirah definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency
AT siahpiranialirezafotuhi definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency
AT mccallasunniegrace definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency
AT zhouambers definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency
AT iyergopal definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency
AT roysushmita definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency
AT sridharanrupa definingreprogrammingcheckpointsfromsinglecellanalysesofinducedpluripotency