Wnt signaling mediates acquisition of blood-brain barrier properties in naïve endothelium derived from human pluripotent stem cells

Abstract
Data availability
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Abstract

Endothelial cells (ECs) in the central nervous system (CNS) acquire their specialized blood-brain barrier (BBB) properties in response to extrinsic signals, with Wnt/β-catenin signaling coordinating multiple aspects of this process. Our knowledge of CNS EC development has been advanced largely by animal models, and human pluripotent stem cells (hPSCs) offer the opportunity to examine BBB development in an in vitro human system. Here we show that activation of Wnt signaling in hPSC-derived naïve endothelial progenitors, but not in matured ECs, leads to robust acquisition of canonical BBB phenotypes including expression of GLUT-1, increased claudin-5, decreased PLVAP and decreased permeability. RNA-seq revealed a transcriptome profile resembling ECs with CNS-like characteristics, including Wnt-upregulated expression of LEF1, APCDD1, and ZIC3. Together, our work defines effects of Wnt activation in naïve ECs and establishes an improved hPSC-based model for interrogation of CNS barriergenesis.

Data availability

RNA-seq data have been deposited in GEO under accession number GSE173206.

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Article and author information

Author details

Benjamin D Gastfriend

Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, United States

Competing interests
Benjamin D Gastfriend, Inventor on a provisional US patent application (63/185815) related to this work..

"This ORCID iD identifies the author of this article:" 0000-0002-4677-1455
Hideaki Nishihara

University of Bern, Bern, Switzerland

Competing interests
Hideaki Nishihara, Inventor on a provisional US patent application (63/185815) related to this work..
Scott G Canfield

Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, United States

Competing interests
No competing interests declared.
Koji L Foreman

Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, United States

Competing interests
No competing interests declared.
Britta Englehardt

University of Bern, Bern, Switzerland

Competing interests
Britta Englehardt, Inventor on a provisional US patent application (63/185815) related to this work..
Sean P Palecek

Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, United States

For correspondence
sppalecek@wisc.edu

Competing interests
Sean P Palecek, Inventor on a provisional US patent application (63/185815) related to this work..
Eric V Shusta

Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, United States

For correspondence
eshusta@wisc.edu

Competing interests
Eric V Shusta, Inventor on a provisional US patent application (63/185815) related to this work..

"This ORCID iD identifies the author of this article:" 0000-0002-4297-0158

Funding

National Institutes of Health (R01 NS103844)

Sean P Palecek
Eric V Shusta

National Institutes of Health (R01 NS107461)

Sean P Palecek
Eric V Shusta

National Institutes of Health (T32 GM008349)

Benjamin D Gastfriend

National Science Foundation (1747503)

Benjamin D Gastfriend

Swiss National Science Foundation (310030_189080)

Britta Englehardt

Bern Center for Precision Medicine

Britta Englehardt

Japan Society for the Promotion of Science (Overseas Research Fellowship)

Hideaki Nishihara

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.