An Induced Pluripotent Stem Cell-Derived Human Blood–Brain Barrier (BBB) Model to Test the Crossing by Adeno-Associated Virus (AAV) Vectors and Antisense Oligonucleotides

Selvakumaran, Jamuna; Ursu, Simona; Bowerman, Melissa; Lu-Nguyen, Ngoc; Wood, Matthew J; Malerba, Alberto; Yáñez-Muñoz, Rafael J

Abstract

The blood-brain barrier (BBB) is the specialised microvasculature system that shields the central nervous system (CNS) from potentially toxic agents. Attempts to develop therapeutic agents targeting the CNS have been hindered by the lack of predictive models of BBB crossing. In vitro models mimicking the human BBB are of great interest, and advances in induced pluripotent stem cell (iPSC) technologies and availability of reproducible differentiation protocols have facilitated progress. In this study we present the efficient differentiation of three different wild-type iPSC lines into brain microvascular endothelial cells (BMECs). Once differentiated, cells displayed several features of BMECs and exhibited significant barrier tightness as measured by trans-endothelial electrical resistance (TEER), ranging from 1500 to >6000 Ωcm 2. To assess the functionality of our BBB models, we analysed the crossing efficiency of adeno-associated virus (AAV) vectors and peptide-conjugated antisense oligonucleotides, both currently used in genetic approaches for the treatment of rare diseases. We demonstrated superior barrier crossing by AAV serotype 9 compared to serotype 8, and no crossing by a cell penetrating peptide-conjugated antisense oligonucleotide. In conclusion, our study shows that iPSC-based models of the human BBB display robust phenotypes and could be used to screen drugs for CNS penetration in culture.

Citation

Selvakumaran, J., Ursu, S., Bowerman, M., Lu-Nguyen, N., Wood, M. J., Malerba, A., & Yáñez-Muñoz, R. J. (2023). An Induced Pluripotent Stem Cell-Derived Human Blood–Brain Barrier (BBB) Model to Test the Crossing by Adeno-Associated Virus (AAV) Vectors and Antisense Oligonucleotides. Biomedicines, 11(10), Article 2700. https://doi.org/10.3390/xxxxx

• Journal Article Type: Article
• Acceptance Date: Sep 28, 2023
• Online Publication Date: Oct 4, 2023
• Publication Date: Oct 4, 2023
• Deposit Date: Sep 28, 2023
• Publicly Available Date: Oct 24, 2023
• Journal: Biomedicines
• Electronic ISSN: 2227-9059
• Publisher: MDPI
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Issue: 10
• Article Number: 2700
• DOI: https://doi.org/10.3390/xxxxx
• Keywords: human blood-brain barrier; brain microvascular endothelial cells; induced pluripotent stem cells; trans-endothelial electrical resistance; gene therapy; spinal muscular atrophy; antisense oligonucleotides; phosphorodiamidate morpholino oligomers; adeno-as