title: Altered long non-coding RNA transcriptomic profiles in brain microvascular endothelium after cerebral ischemia.
abstract: The brain endothelium is an important therapeutic target for the inhibition of cerebrovascular dysfunction in ischemic stroke. Previously, we documented the important regulatory roles of microRNAs in the cerebral vasculature, in particular the cerebral vascular endothelium. However, the functional significance and molecular mechanisms of other classes of non-coding RNAs in the regulation of cerebrovascular endothelial pathophysiology after stroke are completely unknown. Using RNA sequencing (RNA-seq) technology, we profiled long non-coding RNA (lncRNA) expressional signatures in primary brain microvascular endothelial cells (BMECs) after oxygen-glucose deprivation (OGD), an in vitro mimic of ischemic stroke conditions. After 16h of OGD exposure, the expression levels for 362 of the 10,677 lncRNAs analyzed changed significantly, including a total of 147 lncRNAs increased and 70 lncRNAs decreased by more than 2-fold. Among them, the most highly upregulated lncRNAs include Snhg12, Malat1, and lnc-OGD 1006, whereas the most highly downregulated lncRNAs include 281008D09Rik, Peg13, and lnc-OGD 3916. Alteration of the most highly upregulated/downregulated ODG-responsive lncRNAs was further confirmed in cultured BMECs after OGD as well as isolated cerebral microvessels in mice following transient middle cerebral artery occlusion (MCAO) and 24h reperfusion by the quantitative real-time PCR approach. Moreover, promoter analysis of altered ODG-responsive endothelial lncRNA genes by bioinformatics showed substantial transcription factor binding sites on lncRNAs, implying potential transcriptional regulation of those lncRNAs. These findings are the first to identify OGD-responsive brain endothelial lncRNAs, which suggest potential pathological roles for these lncRNAs in mediating endothelial responses to ischemic stimuli. Endothelial-selective lncRNAs may function as a class of novel master regulators in cerebrovascular endothelial pathologies after ischemic stroke.
authors.affiliation: Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, MI 48109, USA.
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, MI 48109, USA.
authors.forename: L
authors.initials: L
authors.lastname: Yuan
authors.affiliation: Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
authors.forename: X
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology, Tulane University School of Medicine, 1430 Tulane Avenue SL83, New Orleans, LA 70112, USA.
authors.forename: M H
authors.initials: MH
authors.lastname: Hamblin
authors.affiliation: Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, MI 48109, USA.
authors.forename: T
authors.initials: T
authors.lastname: Zhu
authors.affiliation: Department of Psychiatry, Molecular and Behavioral Neuroscience Institute, University of Michigan, Ann Arbor, MI 48109, USA.
authors.forename: F
authors.initials: F
authors.lastname: Meng
authors.affiliation: Key Lab of Systems Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Li
authors.affiliation: Cardiovascular Center, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, MI 48109, USA; Department of Cardiac Surgery, University of Michigan Medical Center, Ann Arbor, MI 48109, USA. Electronic address: echenum@umich.edu.
authors.forename: Y E
authors.initials: YE
authors.lastname: Chen
authors.affiliation: Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. Electronic address: yink2@upmc.edu.
authors.forename: K J
authors.initials: KJ
authors.lastname: Yin
mesh_terms: D000818:Animals; D001921:Brain / Q000473:pathology*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D062507:Claudin-5 / Q000378:metabolism; D004195:Disease Models, Animal; D004727:Endothelium / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005947:Glucose / Q000172:deficiency; D000860:Hypoxia / Q000378:metabolism; D020244:Infarction, Middle Cerebral Artery / Q000473:pathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D055806:Microvessels / Q000473:pathology*; D062793:Occludin / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D062085:RNA, Long Noncoding / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: C107466:CLDN5 protein, human; D062507:Claudin-5; D005904:Glial Fibrillary Acidic Protein; D062793:Occludin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D062085:RNA, Long Noncoding; D014157:Transcription Factors; D005947:Glucose
keywords: Brain microvascular endothelial cells; Ischemic stroke; Long non-coding RNAs; Oxygen–glucose deprivation; RNA sequencing; Transcriptome