High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning

Mukul Tewary; Dominika Dziedzicka; Joel Ostblom; Laura Prochazka; Nika Shakiba; Tiam Heydari; Daniel Aguilar-Hidalgo; Curtis Woodford; Elia Piccinini; David Becerra-Alonso; Alice Vickers; Blaise Louis; Nafees Rahman; Davide Danovi; Mieke Geens; Fiona M. Watt; Peter W. Zandstra

doi:10.1371/journal.pbio.3000081

High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning

Mukul Tewary, Dominika Dziedzicka, Joel Ostblom, Laura Prochazka, Nika Shakiba, Tiam Heydari, Daniel Aguilar-Hidalgo, Curtis Woodford, Elia Piccinini, David Becerra-Alonso, Alice Vickers, Blaise Louis, Nafees Rahman, Davide Danovi, Mieke Geens, Fiona M. Watt, Peter W. Zandstra

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Abstract

In vitro models of postimplantation human development are valuable to the fields of regenerative medicine and developmental biology. Here, we report characterization of a robust in vitro platform that enabled high-content screening of multiple human pluripotent stem cell (hPSC) lines for their ability to undergo peri-gastrulation-like fate patterning upon bone morphogenetic protein 4 (BMP4) treatment of geometrically confined colonies and observed significant heterogeneity in their differentiation propensities along a gastrulation associable and neuralization associable axis. This cell line-associated heterogeneity was found to be attributable to endogenous Nodal expression, with up-regulation of Nodal correlated with expression of a gastrulation-associated gene profile, and Nodal down-regulation correlated with a preneurulation-associated gene profile expression. We harness this knowledge to establish a platform of preneurulation-like fate patterning in geometrically confined hPSC colonies in which fates arise because of a BMPs signalling gradient conveying positional information. Our work identifies a Nodal signalling-dependent switch in peri-gastrulation versus preneurulation-associated fate patterning in hPSC cells, provides a technology to robustly assay hPSC differentiation outcomes, and suggests conserved mechanisms of organized fate specification in differentiating epiblast and ectodermal tissues.

Original language	English
Article number	e3000081
Journal	PLoS Biology
Volume	17
Issue number	10
DOIs	https://doi.org/10.1371/journal.pbio.3000081
Publication status	Published - Oct 2019

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Tewary, M., Dziedzicka, D., Ostblom, J., Prochazka, L., Shakiba, N., Heydari, T., Aguilar-Hidalgo, D., Woodford, C., Piccinini, E., Becerra-Alonso, D., Vickers, A., Louis, B., Rahman, N., Danovi, D., Geens, M., Watt, F. M., & Zandstra, P. W. (2019). High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning. PLoS Biology, 17(10), Article e3000081. https://doi.org/10.1371/journal.pbio.3000081

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title = "High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning",

abstract = "In vitro models of postimplantation human development are valuable to the fields of regenerative medicine and developmental biology. Here, we report characterization of a robust in vitro platform that enabled high-content screening of multiple human pluripotent stem cell (hPSC) lines for their ability to undergo peri-gastrulation-like fate patterning upon bone morphogenetic protein 4 (BMP4) treatment of geometrically confined colonies and observed significant heterogeneity in their differentiation propensities along a gastrulation associable and neuralization associable axis. This cell line-associated heterogeneity was found to be attributable to endogenous Nodal expression, with up-regulation of Nodal correlated with expression of a gastrulation-associated gene profile, and Nodal down-regulation correlated with a preneurulation-associated gene profile expression. We harness this knowledge to establish a platform of preneurulation-like fate patterning in geometrically confined hPSC colonies in which fates arise because of a BMPs signalling gradient conveying positional information. Our work identifies a Nodal signalling-dependent switch in peri-gastrulation versus preneurulation-associated fate patterning in hPSC cells, provides a technology to robustly assay hPSC differentiation outcomes, and suggests conserved mechanisms of organized fate specification in differentiating epiblast and ectodermal tissues.",

author = "Mukul Tewary and Dominika Dziedzicka and Joel Ostblom and Laura Prochazka and Nika Shakiba and Tiam Heydari and Daniel Aguilar-Hidalgo and Curtis Woodford and Elia Piccinini and David Becerra-Alonso and Alice Vickers and Blaise Louis and Nafees Rahman and Davide Danovi and Mieke Geens and Watt, {Fiona M.} and Zandstra, {Peter W.}",

year = "2019",

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Tewary, M, Dziedzicka, D, Ostblom, J, Prochazka, L, Shakiba, N, Heydari, T, Aguilar-Hidalgo, D, Woodford, C, Piccinini, E, Becerra-Alonso, D, Vickers, A , Louis, B, Rahman, N, Danovi, D, Geens, M, Watt, FM & Zandstra, PW 2019, 'High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning', PLoS Biology, vol. 17, no. 10, e3000081. https://doi.org/10.1371/journal.pbio.3000081

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T1 - High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning

AU - Tewary, Mukul

AU - Dziedzicka, Dominika

AU - Ostblom, Joel

AU - Prochazka, Laura

AU - Shakiba, Nika

AU - Heydari, Tiam

AU - Aguilar-Hidalgo, Daniel

AU - Woodford, Curtis

AU - Piccinini, Elia

AU - Becerra-Alonso, David

AU - Vickers, Alice

AU - Louis, Blaise

AU - Rahman, Nafees

AU - Danovi, Davide

AU - Geens, Mieke

AU - Watt, Fiona M.

AU - Zandstra, Peter W.

PY - 2019/10

Y1 - 2019/10

N2 - In vitro models of postimplantation human development are valuable to the fields of regenerative medicine and developmental biology. Here, we report characterization of a robust in vitro platform that enabled high-content screening of multiple human pluripotent stem cell (hPSC) lines for their ability to undergo peri-gastrulation-like fate patterning upon bone morphogenetic protein 4 (BMP4) treatment of geometrically confined colonies and observed significant heterogeneity in their differentiation propensities along a gastrulation associable and neuralization associable axis. This cell line-associated heterogeneity was found to be attributable to endogenous Nodal expression, with up-regulation of Nodal correlated with expression of a gastrulation-associated gene profile, and Nodal down-regulation correlated with a preneurulation-associated gene profile expression. We harness this knowledge to establish a platform of preneurulation-like fate patterning in geometrically confined hPSC colonies in which fates arise because of a BMPs signalling gradient conveying positional information. Our work identifies a Nodal signalling-dependent switch in peri-gastrulation versus preneurulation-associated fate patterning in hPSC cells, provides a technology to robustly assay hPSC differentiation outcomes, and suggests conserved mechanisms of organized fate specification in differentiating epiblast and ectodermal tissues.

AB - In vitro models of postimplantation human development are valuable to the fields of regenerative medicine and developmental biology. Here, we report characterization of a robust in vitro platform that enabled high-content screening of multiple human pluripotent stem cell (hPSC) lines for their ability to undergo peri-gastrulation-like fate patterning upon bone morphogenetic protein 4 (BMP4) treatment of geometrically confined colonies and observed significant heterogeneity in their differentiation propensities along a gastrulation associable and neuralization associable axis. This cell line-associated heterogeneity was found to be attributable to endogenous Nodal expression, with up-regulation of Nodal correlated with expression of a gastrulation-associated gene profile, and Nodal down-regulation correlated with a preneurulation-associated gene profile expression. We harness this knowledge to establish a platform of preneurulation-like fate patterning in geometrically confined hPSC colonies in which fates arise because of a BMPs signalling gradient conveying positional information. Our work identifies a Nodal signalling-dependent switch in peri-gastrulation versus preneurulation-associated fate patterning in hPSC cells, provides a technology to robustly assay hPSC differentiation outcomes, and suggests conserved mechanisms of organized fate specification in differentiating epiblast and ectodermal tissues.

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U2 - 10.1371/journal.pbio.3000081

DO - 10.1371/journal.pbio.3000081

M3 - Article

C2 - 31634368

AN - SCOPUS:85074378026

SN - 1544-9173

VL - 17

JO - PLoS Biology

JF - PLoS Biology

IS - 10

M1 - e3000081

ER -

High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning

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