Synergistic induction of blood-brain barrier properties

• 成果类型: Article
• 署名作者: Porkolab, Gergo; Meszaros, Maria; Szecsko, Aniko; Vigh, Judit P.; Walter, Fruzsina R.; Figueiredo, Ricardo; Kalomista, Ildiko; Hoyk, Zsofia; Vizsnyiczai, Gaszton; Grof, Ilona; Jan, Jeng-Shiung; Gosselet, Fabien; Pirity, Melinda K.; Vastag, Monika; Hudson, Natalie; Campbell, Matthew; Veszelka, Szilvia; Deli, Maria A.
• 署名单位: HUN-REN; HUN-REN Biological Research Center; Institute of Biophysics - HAS; Szeged University; Gedeon Richter Chemistry Works; National Cheng Kung University; Universite d'Artois; HUN-REN; HUN-REN Biological Research Center; Institute of Genetics - HAS; Trinity College Dublin
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14429
• DOI: 10.1073/pnas.2316006121
• 发表日期: 2024-05-21
• 关键词: endothelial-cells; pericytes; culture; MODEL; cns; establishment; transporters; astrocytes; disease

摘要

Blood-brain barrier (BBB) models derived from human stem cells are powerful tools to improve our understanding of cerebrovascular diseases and to facilitate drug development for the human brain. Yet providing stem cell-derived endothelial cells with the right signaling cues to acquire BBB characteristics while also retaining their vascular identity remains challenging. Here, we show that the simultaneous activation of cyclic AMP and Wnt/beta-catenin signaling and inhibition of the TGF-beta pathway in endothelial cells robustly induce BBB properties in vitro. To target this interaction, we present a small-molecule cocktail named cARLA, which synergistically enhances barrier tightness in a range of BBB models across species. Mechanistically, we reveal that the three pathways converge on Wnt/beta-catenin signaling to mediate the effect of cARLA via the tight junction protein claudin-5. We demonstrate that cARLA shifts the gene expressional profile of human stem cell-derived endothelial cells toward the in vivo brain endothelial signature, with a higher glycocalyx density and efflux pump activity, lower rates of endocytosis, and a characteristic endothelial response to proinflammatory cytokines. Finally, we illustrate how cARLA can improve the predictive value of human BBB models regarding the brain penetration of drugs and targeted nanoparticles. Due to its synergistic effect, high reproducibility, and ease of use, cARLA has the potential to advance drug development for the human brain by improving BBB models across laboratories.