Blocking lncRNA H19-miR-19a-Id2 axis attenuates hypoxia/ischemia induced neuronal injury.

Zhipeng Xiao,Jing Cui,Yongming Qiu,Liemei Guo,Rogelio Medina,Zezhi Li,Yingying Lin,Jared S Rosenblum,Xiaohua Zhang,Sophie Zhuang

doi:10.18632/aging.101999

Abstract

Elevated expression of lncRNA H19 (H19) in the setting of hypoxia has been implicated as a promising therapeutic target for various cancers. However, little is known about the impact and underlying mechanism of H19 in ischemic brain stroke. This study found that H19 levels were elevated in the serum of stroke patients, as well as in the ischemic penumbra of rats with middle cerebral artery occlusion/reperfusion (MCAO/R) injury and neuronal cells with oxygen glucose deprivation (OGD). Further, knockdown of H19 with siRNA alleviated cell apoptosis in OGD neuronal cells, and inhibition of H19 in MCAO/R rats significantly decreased neurological deficit, brain infarct volume and neuronal apoptosis. Lastly, with gain and loss of function studies, dual luciferase reported assay, RNA immunoprecipitation (RIP) and pull-down experiments, we demonstrated the dual competitive interaction of miR-19a with H19 and the 3’-UTR of Id2 mRNA, resulting in the identification of the H19-miR-19a-Id2 axis. With biological studies, we also revealed that H19-miR-19a-Id2 axis modulated hypoxia induced neuronal apoptosis. This study demonstrates that the identified H19-miR-19a-Id2 axis plays a critical role in hypoxia induced neuronal apoptosis, and blocking this axis may serve as a novel therapeutic strategy for ischemic brain injury.

Highlights

Cerebral ischemic stroke continues to incur high morbidity and mortality rates [1]
Quantitative real-time polymerase chain reaction revealed that H19 was expressed at low level in the plasma of normal control patients, but it was significantly upregulated in ischemic stroke patients (Figure 1A; p
We first demonstrated that H19 level was significantly elevated in ischemic stroke patients, in-vivo middle cerebral artery occlusion/reperfusion (MCAO/R) animal model and in-vitro oxygen glucose deprivation (OGD) neuronal cell model

Summary

Introduction

Cerebral ischemic stroke continues to incur high morbidity and mortality rates [1]. Therapeutic protocols have continued to evolve as our understanding of the umbra and salvageable penumbra increases. These advances have been broadly aimed at either 1) resolving the infarct in a timely fashion to limit the continued damage incurred by hypoxic stress and resultant neuronal injury or 2) stimulating neural replacement [2]. The application of molecular understanding of hypoxic neuronal injury has been limited [3, 4]. Absence of an Id2 targeted inhibitor, has limited its application as a potential therapeutic option to protect brain from ischemic injury. Seeking the upstream regulating molecules of Id2 is of great significance

Methods

Results

Conclusion