Hypoxia-Induced MicroRNA-212/132 Alter Blood-Brain Barrier Integrity Through Inhibition of Tight Junction-Associated Proteins in Human and Mouse Brain Microvascular Endothelial Cells

Overview

Journal Transl Stroke Res

Publisher Springer

Date 2019 Jan 9

PMID 30617994

Citations 72

Authors

Malgorzata Burek

Anna Konig

Mareike Lang

Jan Fiedler

Sabrina Oerter

Norbert Roewer

Michael Bohnert

Serge C Thal

Kinga G Blecharz-Lang

Johannes Woitzik

Thomas Thum

Carola Y Forster

Affiliations

Soon will be listed here.

Abstract

Blood-brain barrier (BBB) integrity is one of the important elements of central nervous system (CNS) homeostasis. MicroRNAs (miRs) have been demonstrated to play a role in many CNS disorders such as stroke and traumatic brain injury. MiR-212/132 are highly expressed in the CNS but their role at the BBB has not been characterized yet. Thus, we analyzed the expression of miR-212/132 in hypoxic mouse and human brain microvascular endothelial cells (BMEC) as well as in posttraumatic mouse and human brain tissue and serum exosomes. MiR-212/132 expression was detected in brain capillaries by in situ hybridization and was increased up to ten times in hypoxic BMEC. Over-expression of pre-miR-212/132 in BMEC decreased barrier properties and reduced migration of BMEC in the wound healing assay. We identified and validated tight junction proteins claudin-1 (Cldn1), junctional adhesion molecule 3 (Jam3), and tight junction-associated protein 1 (Tjap1) as potential miR-212/132 targets. Over-expression of miRs led to a decrease in mRNA and protein expression of Cldn1, Jam3, and Tjap1, which could be rescued by a respective anti-miR. In conclusion, our study identifies miR-212/132 as critical players at the hypoxic BBB. In addition, we propose three new direct miR-212/132 targets to be involved in miR-212/132-mediated effects on BBB properties.

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References

Orlova V, Economopoulou M, Lupu F, Santoso S, Chavakis T . Junctional adhesion molecule-C regulates vascular endothelial permeability by modulating VE-cadherin-mediated cell-cell contacts. J Exp Med. 2006; 203(12):2703-14. PMC: 2118160. DOI: 10.1084/jem.20051730. View

Fletcher N, Wilson G, Murray J, Hu K, Lewis A, Reynolds G . Hepatitis C virus infects the endothelial cells of the blood-brain barrier. Gastroenterology. 2011; 142(3):634-643.e6. PMC: 3801216. DOI: 10.1053/j.gastro.2011.11.028. View

Ucar A, Vafaizadeh V, Jarry H, Fiedler J, Klemmt P, Thum T . miR-212 and miR-132 are required for epithelial stromal interactions necessary for mouse mammary gland development. Nat Genet. 2010; 42(12):1101-8. DOI: 10.1038/ng.709. View

Fiedler J, Jazbutyte V, Kirchmaier B, Gupta S, Lorenzen J, Hartmann D . MicroRNA-24 regulates vascularity after myocardial infarction. Circulation. 2011; 124(6):720-30. DOI: 10.1161/CIRCULATIONAHA.111.039008. View

Tognini P, Pizzorusso T . MicroRNA212/132 family: molecular transducer of neuronal function and plasticity. Int J Biochem Cell Biol. 2011; 44(1):6-10. DOI: 10.1016/j.biocel.2011.10.015. View

Yanev P, Seevinck P, Rudrapatna U, Bouts M, van der Toorn A, Gertz K . Magnetic resonance imaging of local and remote vascular remodelling after experimental stroke. J Cereb Blood Flow Metab. 2016; 37(8):2768-2779. PMC: 5536787. DOI: 10.1177/0271678X16674737. View

Stankowski J, Gupta R . Therapeutic targets for neuroprotection in acute ischemic stroke: lost in translation?. Antioxid Redox Signal. 2010; 14(10):1841-51. PMC: 3120088. DOI: 10.1089/ars.2010.3292. View

Mathers C, Loncar D . Projections of global mortality and burden of disease from 2002 to 2030. PLoS Med. 2006; 3(11):e442. PMC: 1664601. DOI: 10.1371/journal.pmed.0030442. View

Timaru-Kast R, Herbig E, Luh C, Engelhard K, Thal S . Influence of Age on Cerebral Housekeeping Gene Expression for Normalization of Quantitative Polymerase Chain Reaction after Acute Brain Injury in Mice. J Neurotrauma. 2015; 32(22):1777-88. DOI: 10.1089/neu.2014.3784. View

10.

Blecharz-Lang K, Prinz V, Burek M, Frey D, Schenkel T, Krug S . Gelatinolytic activity of autocrine matrix metalloproteinase-9 leads to endothelial de-arrangement in Moyamoya disease. J Cereb Blood Flow Metab. 2018; 38(11):1940-1953. PMC: 6259317. DOI: 10.1177/0271678X18768443. View

11.

Kleinschnitz C, Blecharz K, Kahles T, Schwarz T, Kraft P, Gobel K . Glucocorticoid insensitivity at the hypoxic blood-brain barrier can be reversed by inhibition of the proteasome. Stroke. 2011; 42(4):1081-9. DOI: 10.1161/STROKEAHA.110.592238. View

12.

Burek M, Arias-Loza P, Roewer N, Forster C . Claudin-5 as a novel estrogen target in vascular endothelium. Arterioscler Thromb Vasc Biol. 2009; 30(2):298-304. DOI: 10.1161/ATVBAHA.109.197582. View

13.

Wilhelm I, Fazakas C, Molnar K, Vegh A, Hasko J, Krizbai I . Foe or friend? Janus-faces of the neurovascular unit in the formation of brain metastases. J Cereb Blood Flow Metab. 2017; 38(4):563-587. PMC: 5888855. DOI: 10.1177/0271678X17732025. View

14.

Li X, Li D, Wikstrom J, Pivarcsi A, Sonkoly E, Stahle M . MicroRNA-132 promotes fibroblast migration via regulating RAS p21 protein activator 1 in skin wound healing. Sci Rep. 2017; 7(1):7797. PMC: 5552762. DOI: 10.1038/s41598-017-07513-0. View

15.

Fujibe M, Chiba H, Kojima T, Soma T, Wada T, Yamashita T . Thr203 of claudin-1, a putative phosphorylation site for MAP kinase, is required to promote the barrier function of tight junctions. Exp Cell Res. 2004; 295(1):36-47. DOI: 10.1016/j.yexcr.2003.12.014. View

16.

Dilling C, Roewer N, Forster C, Burek M . Multiple protocadherins are expressed in brain microvascular endothelial cells and might play a role in tight junction protein regulation. J Cereb Blood Flow Metab. 2017; 37(10):3391-3400. PMC: 5624389. DOI: 10.1177/0271678X16688706. View

17.

Weksler B, Subileau E, Perriere N, Charneau P, Holloway K, Leveque M . Blood-brain barrier-specific properties of a human adult brain endothelial cell line. FASEB J. 2005; 19(13):1872-4. DOI: 10.1096/fj.04-3458fje. View

18.

Yao C, Shi X, Zhang Z, Zhou S, Qian T, Wang Y . Hypoxia-Induced Upregulation of miR-132 Promotes Schwann Cell Migration After Sciatic Nerve Injury by Targeting PRKAG3. Mol Neurobiol. 2015; 53(8):5129-39. DOI: 10.1007/s12035-015-9449-y. View

19.

Wyss L, Schafer J, Liebner S, Mittelbronn M, Deutsch U, Enzmann G . Junctional adhesion molecule (JAM)-C deficient C57BL/6 mice develop a severe hydrocephalus. PLoS One. 2012; 7(9):e45619. PMC: 3445510. DOI: 10.1371/journal.pone.0045619. View

20.

Burek M, Forster C . Cloning and characterization of the murine claudin-5 promoter. Mol Cell Endocrinol. 2008; 298(1-2):19-24. DOI: 10.1016/j.mce.2008.09.041. View