The structural and functional abnormalities in the cerebral endothelium during cerebral ischemia are well- known to affect the extent of ischemic brain injury and neurological outcomes. Therefore, the brain endothelium becomes an important therapeutic target for the inhibition of cerebrovascular dysfunction in ischemic stroke. However, the regulatory mechanisms of cerebral endothelial dysfunction after cerebral ischemia remain largely unexplored. Previously we have 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 ischemic stroke are completely unknown. Using RNA-sequencing technology, we profiled long non-coding RNAs (lncRNAs) expressional signatures in primary brain microvascular endothelial cells (BMECs) after oxygen-glucose deprivation (OGD). 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. Interestingly, metastasis associated lung adenocarcinoma transcript 1 (Malat1) is one of the most highly upregulated OGD-responsive endothelial lncRNAs, which has been initially discovered as a tumor-associated lncRNA involved in epigenetic control of gene expression. Furthermore, we found that loss-of-Malat1 function significantly increased OGD- induced loss of endothelial marker and endothelial inflammation in BMECs. In this exploratory proposal, we will test our central hypothesis that Malat1 plays a critical role in protecting the cerebral microvasculature from cerebral ischemic insults by inhibiting endothelial degeneration and inflammation. We propose to test this hypothesis with the following two aims:
Aim 1 is to define the pathological profiles of Malat1 in cerebrovascular endothelium after ischemic stroke.
Aim 2 is to identify the functional role of Malat1 in regulating ischemic cerebrovascular and brain damage. The broad, long-term goal is to understand the essential role of lncRNAs in cerebrovascular pathologies after ischemic stroke and to develop potential lncRNA-based therapies/ intervention for ischemic stroke.

Public Health Relevance

Stroke is the fourth most common cause of death and the leading cause of adult disability in the United States. Currently, acute therapeutic interventions are limited to thrombolytic therapy within a narrow time window and development of effective therapies is urgently required. The object of this application is to test our hypothesis that long non-coding RNAs (lncRNAs) play a critical role in protecting the cerebral microvasculature from cerebral ischemic insults by inhibition of endothelial degeneration and inflammation. The successful implementation of this exploratory proposal will lead us to discover novel cerebral endothelial regulators and may reveal new lncRNA-based approaches for therapeutic intervention in cerebrovascular diseases such as stroke.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21NS094930-03
Application #
9121632
Study Section
Neural Oxidative Metabolism and Death Study Section (NOMD)
Program Officer
Koenig, James I
Project Start
2015-11-11
Project End
2017-07-31
Budget Start
2016-08-01
Budget End
2017-07-31
Support Year
3
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Pittsburgh
Department
Neurology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Sun, Ping; Liu, Da Zhi; Jickling, Glen C et al. (2018) MicroRNA-based therapeutics in central nervous system injuries. J Cereb Blood Flow Metab 38:1125-1148
Zhang, Xuejing; Hamblin, Milton H; Yin, Ke-Jie (2018) Noncoding RNAs and Stroke. Neuroscientist :1073858418769556
Tan, Zhou; Li, Jingya; Zhang, Xuejing et al. (2018) P53 Promotes Retinoid Acid-induced Smooth Muscle Cell Differentiation by Targeting Myocardin. Stem Cells Dev 27:534-544
Tang, Xuelian; Liu, Kai; Hamblin, Milton H et al. (2018) Genetic Deletion of Krüppel-Like Factor 11 Aggravates Ischemic Brain Injury. Mol Neurobiol 55:2911-2921
Zhang, Xuejing; Tang, Xuelian; Liu, Kai et al. (2017) Long Noncoding RNA Malat1 Regulates Cerebrovascular Pathologies in Ischemic Stroke. J Neurosci 37:1797-1806
Zhang, Xuejing; Hamblin, Milton H; Yin, Ke-Jie (2017) The long noncoding RNA Malat1: Its physiological and pathophysiological functions. RNA Biol 14:1705-1714
Yang, Xinxin; Tang, Xuelian; Sun, Ping et al. (2017) MicroRNA-15a/16-1 Antagomir Ameliorates Ischemic Brain Injury in Experimental Stroke. Stroke 48:1941-1947
Zhang, J; Yuan, L; Zhang, X et al. (2016) Altered long non-coding RNA transcriptomic profiles in brain microvascular endothelium after cerebral ischemia. Exp Neurol 277:162-170