P450 oxidoreductase regulates barrier maturation by mediating retinoic acid metabolism in a model of the human BBB

Dor Zlotnik; Tatiana Rabinski; Aviv Halfon; Shira Anzi; Inbar Plaschkes; Hadar Benyamini; Yuval Nevo; Orly Yahalom Gershoni; Benyamin Rosental; Eli Hershkovitz; Ayal Ben-Zvi; Gad D. Vatine

doi:10.1016/j.stemcr.2022.07.010

P450 oxidoreductase regulates barrier maturation by mediating retinoic acid metabolism in a model of the human BBB

Dor Zlotnik, Tatiana Rabinski, Aviv Halfon, Shira Anzi, Inbar Plaschkes, Hadar Benyamini, Yuval Nevo, Orly Yahalom Gershoni, Benyamin Rosental, Eli Hershkovitz, Ayal Ben-Zvi, Gad D. Vatine^*

^*Corresponding author for this work

Department of Developmental Biology and Cancer Research

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

The blood-brain barrier (BBB) selectively regulates the entry of molecules into the central nervous system (CNS). A crosstalk between brain microvascular endothelial cells (BMECs) and resident CNS cells promotes the acquisition of functional tight junctions (TJs). Retinoic acid (RA), a key signaling molecule during embryonic development, is used to enhance in vitro BBB models’ functional barrier properties. However, its physiological relevance and affected pathways are not fully understood. P450 oxidoreductase (POR) regulates the enzymatic activity of microsomal cytochromes. POR-deficient (PORD) patients display impaired steroid homeostasis and cognitive disabilities. Here, we used both patient-specific POR-deficient and CRISPR-Cas9-mediated POR-depleted induced pluripotent stem cell (iPSC)-derived BMECs (iBMECs) to study the role of POR in the acquisition of functional barrier properties. We demonstrate that POR regulates cellular RA homeostasis and that POR deficiency leads to the accumulation of RA within iBMECs, resulting in the impaired acquisition of TJs and, consequently, to dysfunctional development of barrier properties.

Original language	American English
Pages (from-to)	2050-2063
Number of pages	14
Journal	Stem Cell Reports
Volume	17
Issue number	9
DOIs	https://doi.org/10.1016/j.stemcr.2022.07.010
State	Published - 13 Sep 2022

Bibliographical note

Funding Information:
This work was supported by grants from the Israel Science Foundation (ISF), Israel grants 1621/18 and 2327/18 and the Ministry of Science and Technology (MoST), Israel grant 3-15647 to G.D.V. We thank Dr. Abed Nasereddin from the Genomic Applications Laboratory, The Core Research Facility, The Faculty of Medicine, HUJI for support in RNA-seq experiments. The graphic abstract was created with BioRender.com .

Funding Information:
This work was supported by grants from the Israel Science Foundation (ISF), Israel grants 1621/18 and 2327/18 and the Ministry of Science and Technology (MoST), Israel grant 3-15647 to G.D.V. We thank Dr. Abed Nasereddin from the Genomic Applications Laboratory, The Core Research Facility, The Faculty of Medicine, HUJI for support in RNA-seq experiments. The graphic abstract was created with BioRender.com. The authors declare no conflict of interest.

Publisher Copyright:
© 2022 The Authors

Keywords

BBB
P450 oxidoreductase
POR
blood-brain barrier
cytochrome
disease modeling
iPSCs
metabolism
retinoic acid
tight junctions

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10.1016/j.stemcr.2022.07.010

Cite this

Zlotnik, D., Rabinski, T., Halfon, A., Anzi, S., Plaschkes, I., Benyamini, H., Nevo, Y., Gershoni, O. Y., Rosental, B., Hershkovitz, E., Ben-Zvi, A., & Vatine, G. D. (2022). P450 oxidoreductase regulates barrier maturation by mediating retinoic acid metabolism in a model of the human BBB. Stem Cell Reports, 17(9), 2050-2063. https://doi.org/10.1016/j.stemcr.2022.07.010

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abstract = "The blood-brain barrier (BBB) selectively regulates the entry of molecules into the central nervous system (CNS). A crosstalk between brain microvascular endothelial cells (BMECs) and resident CNS cells promotes the acquisition of functional tight junctions (TJs). Retinoic acid (RA), a key signaling molecule during embryonic development, is used to enhance in vitro BBB models{\textquoteright} functional barrier properties. However, its physiological relevance and affected pathways are not fully understood. P450 oxidoreductase (POR) regulates the enzymatic activity of microsomal cytochromes. POR-deficient (PORD) patients display impaired steroid homeostasis and cognitive disabilities. Here, we used both patient-specific POR-deficient and CRISPR-Cas9-mediated POR-depleted induced pluripotent stem cell (iPSC)-derived BMECs (iBMECs) to study the role of POR in the acquisition of functional barrier properties. We demonstrate that POR regulates cellular RA homeostasis and that POR deficiency leads to the accumulation of RA within iBMECs, resulting in the impaired acquisition of TJs and, consequently, to dysfunctional development of barrier properties.",

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note = "Funding Information: This work was supported by grants from the Israel Science Foundation (ISF), Israel grants 1621/18 and 2327/18 and the Ministry of Science and Technology (MoST), Israel grant 3-15647 to G.D.V. We thank Dr. Abed Nasereddin from the Genomic Applications Laboratory, The Core Research Facility, The Faculty of Medicine, HUJI for support in RNA-seq experiments. The graphic abstract was created with BioRender.com . Funding Information: This work was supported by grants from the Israel Science Foundation (ISF), Israel grants 1621/18 and 2327/18 and the Ministry of Science and Technology (MoST), Israel grant 3-15647 to G.D.V. We thank Dr. Abed Nasereddin from the Genomic Applications Laboratory, The Core Research Facility, The Faculty of Medicine, HUJI for support in RNA-seq experiments. The graphic abstract was created with BioRender.com. The authors declare no conflict of interest. Publisher Copyright: {\textcopyright} 2022 The Authors",

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AU - Rabinski, Tatiana

AU - Halfon, Aviv

AU - Anzi, Shira

AU - Plaschkes, Inbar

AU - Benyamini, Hadar

AU - Nevo, Yuval

AU - Gershoni, Orly Yahalom

AU - Rosental, Benyamin

AU - Hershkovitz, Eli

AU - Ben-Zvi, Ayal

AU - Vatine, Gad D.

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N2 - The blood-brain barrier (BBB) selectively regulates the entry of molecules into the central nervous system (CNS). A crosstalk between brain microvascular endothelial cells (BMECs) and resident CNS cells promotes the acquisition of functional tight junctions (TJs). Retinoic acid (RA), a key signaling molecule during embryonic development, is used to enhance in vitro BBB models’ functional barrier properties. However, its physiological relevance and affected pathways are not fully understood. P450 oxidoreductase (POR) regulates the enzymatic activity of microsomal cytochromes. POR-deficient (PORD) patients display impaired steroid homeostasis and cognitive disabilities. Here, we used both patient-specific POR-deficient and CRISPR-Cas9-mediated POR-depleted induced pluripotent stem cell (iPSC)-derived BMECs (iBMECs) to study the role of POR in the acquisition of functional barrier properties. We demonstrate that POR regulates cellular RA homeostasis and that POR deficiency leads to the accumulation of RA within iBMECs, resulting in the impaired acquisition of TJs and, consequently, to dysfunctional development of barrier properties.

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JF - Stem Cell Reports

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P450 oxidoreductase regulates barrier maturation by mediating retinoic acid metabolism in a model of the human BBB

Abstract

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Keywords

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