Wnt signaling mediates acquisition of blood-brain barrier properties in naïve endothelium derived from human pluripotent stem cells
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.
-
Transcriptional and Epigenomic Landscapes of CNS and non-CNS Vascular Endothelial CellsGSM3040844 GSM3040845 GSM3040852 GSM3040853 GSM3040858 GSM3040859 GSM3040864 GSM3040865.
-
Gene expression profiles of liver and lung endothelial cells during normal and upregulation of Wnt/beta-catenin signalingGSM2498580 GSM2498581 GSM2498582 GSM2498583 GSM2498584 GSM2498585 GSM2498586 GSM2498587.
-
The role of beta-catenin signaling in regulating barrier vs. non-barrier gene expression programs in circumventricular organ and ocular vasculaturesGSM3455653 GSM3455654 GSM3455657 GSM3455658 GSM3455661 GSM3455662 GSM3455665 GSM3455666.
-
A genome-wide view of the de-differentiation of central nervous system endothelial cells in cultureGSM4160534 GSM4160535 GSM4160536 GSM4160537 GSM4160538 GSM4160539 GSM4160540 GSM4160541 GSM4160542 GSM4160543.
-
Construction of A Human Cell Landscape by Single-cell mRNA-seqGSM3980129 GSM4008656 GSM4008657 GSM4008658.
Article and author information
Author details
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.
Reviewing Editor
- Stefan Liebner, University Hospital, Goethe University Frankfurt, Germany
Version history
- Preprint posted: May 14, 2021 (view preprint)
- Received: June 4, 2021
- Accepted: November 9, 2021
- Accepted Manuscript published: November 10, 2021 (version 1)
- Accepted Manuscript updated: November 12, 2021 (version 2)
- Version of Record published: December 10, 2021 (version 3)
Copyright
© 2021, Gastfriend et al.
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.
Metrics
-
- 3,195
- Page views
-
- 424
- Downloads
-
- 22
- Citations
Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.
Download links
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Further reading
-
- Developmental Biology
- Neuroscience
Human fetal development has been associated with brain health at later stages. It is unknown whether growth in utero, as indexed by birth weight (BW), relates consistently to lifespan brain characteristics and changes, and to what extent these influences are of a genetic or environmental nature. Here we show remarkably stable and lifelong positive associations between BW and cortical surface area and volume across and within developmental, aging and lifespan longitudinal samples (N = 5794, 4–82 y of age, w/386 monozygotic twins, followed for up to 8.3 y w/12,088 brain MRIs). In contrast, no consistent effect of BW on brain changes was observed. Partly environmental effects were indicated by analysis of twin BW discordance. In conclusion, the influence of prenatal growth on cortical topography is stable and reliable through the lifespan. This early-life factor appears to influence the brain by association of brain reserve, rather than brain maintenance. Thus, fetal influences appear omnipresent in the spacetime of the human brain throughout the human lifespan. Optimizing fetal growth may increase brain reserve for life, also in aging.
-
- Developmental Biology
- Immunology and Inflammation
During embryogenesis, the fetal liver becomes the main hematopoietic organ, where stem and progenitor cells as well as immature and mature immune cells form an intricate cellular network. Hematopoietic stem cells (HSCs) reside in a specialized niche, which is essential for their proliferation and differentiation. However, the cellular and molecular determinants contributing to this fetal HSC niche remain largely unknown. Macrophages are the first differentiated hematopoietic cells found in the developing liver, where they are important for fetal erythropoiesis by promoting erythrocyte maturation and phagocytosing expelled nuclei. Yet, whether macrophages play a role in fetal hematopoiesis beyond serving as a niche for maturing erythroblasts remains elusive. Here, we investigate the heterogeneity of macrophage populations in the murine fetal liver to define their specific roles during hematopoiesis. Using a single-cell omics approach combined with spatial proteomics and genetic fate-mapping models, we found that fetal liver macrophages cluster into distinct yolk sac-derived subpopulations and that long-term HSCs are interacting preferentially with one of the macrophage subpopulations. Fetal livers lacking macrophages show a delay in erythropoiesis and have an increased number of granulocytes, which can be attributed to transcriptional reprogramming and altered differentiation potential of long-term HSCs. Together, our data provide a detailed map of fetal liver macrophage subpopulations and implicate macrophages as part of the fetal HSC niche.