The regulatory architecture of the primed pluripotent cell state

Abstract Despite extensive research, the gene regulatory architecture governing mammalian cell states remains poorly understood. Here we present an integrative systems biology approach to elucidate the network architecture of primed state pluripotency. Using an unbiased methodology, we identified an...

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Main Authors: Bo I. Li, Mariano J. Alvarez, Hui Zhao, Napon Chirathivat, Andrea Califano, Michael M. Shen
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-57894-4
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author Bo I. Li
Mariano J. Alvarez
Hui Zhao
Napon Chirathivat
Andrea Califano
Michael M. Shen
author_facet Bo I. Li
Mariano J. Alvarez
Hui Zhao
Napon Chirathivat
Andrea Califano
Michael M. Shen
author_sort Bo I. Li
collection DOAJ
description Abstract Despite extensive research, the gene regulatory architecture governing mammalian cell states remains poorly understood. Here we present an integrative systems biology approach to elucidate the network architecture of primed state pluripotency. Using an unbiased methodology, we identified and experimentally confirmed 132 transcription factors as master regulators (MRs) of mouse epiblast stem cell (EpiSC) pluripotency, many of which were further validated by CRISPR-mediated functional assays. To assemble a comprehensive regulatory network, we silenced each of the 132 MRs to assess their effects on the other MRs and their transcriptional targets, yielding a network of 1273 MR → MR interactions. Network architecture analyses revealed four functionally distinct MR modules (communities), and identified key Speaker and Mediator MRs based on their hierarchical rank and centrality. Our findings elucidate the de-centralized logic of a “communal interaction” model in which the balanced activities of four MR communities maintain primed state pluripotency.
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spelling doaj-art-a2eaf6f64f41436cb99126d62e4e18cc2025-08-20T02:11:47ZengNature PortfolioNature Communications2041-17232025-04-0116111910.1038/s41467-025-57894-4The regulatory architecture of the primed pluripotent cell stateBo I. Li0Mariano J. Alvarez1Hui Zhao2Napon Chirathivat3Andrea Califano4Michael M. Shen5Department of MedicineSystems BiologyDepartment of MedicineDepartment of MedicineDepartment of MedicineDepartment of MedicineAbstract Despite extensive research, the gene regulatory architecture governing mammalian cell states remains poorly understood. Here we present an integrative systems biology approach to elucidate the network architecture of primed state pluripotency. Using an unbiased methodology, we identified and experimentally confirmed 132 transcription factors as master regulators (MRs) of mouse epiblast stem cell (EpiSC) pluripotency, many of which were further validated by CRISPR-mediated functional assays. To assemble a comprehensive regulatory network, we silenced each of the 132 MRs to assess their effects on the other MRs and their transcriptional targets, yielding a network of 1273 MR → MR interactions. Network architecture analyses revealed four functionally distinct MR modules (communities), and identified key Speaker and Mediator MRs based on their hierarchical rank and centrality. Our findings elucidate the de-centralized logic of a “communal interaction” model in which the balanced activities of four MR communities maintain primed state pluripotency.https://doi.org/10.1038/s41467-025-57894-4
spellingShingle Bo I. Li
Mariano J. Alvarez
Hui Zhao
Napon Chirathivat
Andrea Califano
Michael M. Shen
The regulatory architecture of the primed pluripotent cell state
Nature Communications
title The regulatory architecture of the primed pluripotent cell state
title_full The regulatory architecture of the primed pluripotent cell state
title_fullStr The regulatory architecture of the primed pluripotent cell state
title_full_unstemmed The regulatory architecture of the primed pluripotent cell state
title_short The regulatory architecture of the primed pluripotent cell state
title_sort regulatory architecture of the primed pluripotent cell state
url https://doi.org/10.1038/s41467-025-57894-4
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