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, and decreased PLVAP. 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.

Therapeutic modulation of the blood-brain barrier and ischemic stroke by a bioengineered FZD4-selective WNT surrogate

“…3a-c). Plasmalemma vesicle-associated protein (PLVAP) is an EC fenestration component that regulates BBB/BRB permeability and is upregulated in pathological conditions associated with compromised barrier function and is decreased by WNT/β-catenin signaling 48,53,54 . Accordingly, Plvap was an inferred MR that was indeed overexpressed in Ndp KO versus WT mice (Fig.…”

Three-dimensional microenvironment regulates gene expression, function, and tight junction dynamics of iPSC-derived blood-brain barrier microvessels

“…S5B ). To characterize the magnitude of gene expression changes induced by 3D microenvironment, we compared Spearman correlation coefficients across recent studies of shear stress, chemical induction, and TF reprogramming [23, 25, 26, 28] ( Fig. S5C-D ).…”

“…S5C-D ). The magnitude of gene expression changes induced by the 3D microenvironment (ρ = 0.984 between 3D and 2D iBMECs) was similar to other perturbations explored to induce brain endothelial identity, including shear stress (ρ = 0.980 between 2.4 dyne cm -2 and static iBMEC monolayers) [28], agonism of Wnt/β-catenin signaling in endothelial progenitor cells (ρ = 0.977 between EPCs exposed to CHIR99021 or DMSO) [26], and inhibition of TGFβ receptor in iECs (ρ = 0.983 between iECs exposed to RepSox or DMSO) [25] ( Fig. S5D ).…”

“…While recent evidence suggests that iBMECs possess a component of epithelial identity [23], these cells remain a critical source for BBB modeling as they exhibit functional barrier characteristics including high TEER, low permeability, efflux activity, and nutrient transport, while enabling highly scalable and patient-specific experimentation [1,17]. Recent approaches to drive brain endothelial identity by transcription factor (TF) reprogramming of iPSC-derived cells [23,24] or by chemical exposure to Wnt agonists/ligands or TGF-beta inhibitors [25,26] hold promise, but have been unable to achieve physiological barrier function with TEER values typically < 200 Ω cm 2 .…”

Establishment of a novel amyotrophic lateral sclerosis patient-derived blood-brain barrier model: Investigating barrier dysfunction and immune cell interaction

“…Second, they lack several cell surface adhesion molecules such as selectins, ICAM-2, and VCAM-1, which is essential to studying immune cell interaction with the BBB (Nishihara et al, 2020). Our hiPSC-derived BBB model does not show epithelial genes and possesses both proper tight junction integrity and robust expression of adhesion molecules essential for immune cell interaction with the BBB, making this model unique and suitable for studying the link between the peripheral immune system and the CNS via the BBB (Nishihara et al, 2020; Gastfriend et al, 2021).…”