Novel long non-coding RNAs in neuronal survival in focal cerebral ischemia

新型长非编码RNA对局灶性脑缺血神经元存活的影响

基本信息

批准号：
9230451
负责人：
Qiming Jane Wang
金额：
$ 23.25万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9230451
关键词：
Affect Alteplase Animal Model Biological Process Brain Brain Injuries Brain Ischemia Cause of Death Cell Culture Techniques Cell Death Cell model Cells Cerebral Ischemia Development Disease Down-Regulation Event Family Genetic Transcription Glucose Goals I Kappa B-Alpha In Vitro Injury Ischemia Ischemic Brain Injury Ischemic Stroke Length Light Maps Mediating Messenger RNA Modeling Molecular Mus Neuraxis Neuronal Injury Neurons Nucleotides Operative Surgical Procedures Oxygen Pathologic Processes Pattern Pharmacology Phosphotransferases Pilot Projects Play Protein Family Protein Kinase Proteins Rattus Recovery Reperfusion Therapy Role Signal Pathway Signal Transduction Site Slice Small Interfering RNA Stroke Testing Therapeutic Intervention Time Transcript Transcriptional Regulation United States Untranslated RNA base calmodulin-dependent protein kinase II deprivation disability effective therapy in vivo insight knock-down mRNA Stability mouse model neuronal survival neuroprotection new therapeutic target novel public health relevance relating to nervous system response stroke intervention targeted treatment therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Focal cerebral ischemia (ischemic stroke) is the most common cause of disability and the fourth leading cause of death in the United State. Despite of the efforts on developing the pharmacological and surgical treatments of the disease, tissue plasminogen activator (tPA) is the only effective therapy at present. A better understanding of the pathological process and the discovery of new targets and therapies will significantly advance the field. There have been little studies on lncRNAs in cerebral ischemia. Through an lncRNA array analysis in a rat model of focal cerebral ischemia/reperfusion (IR), we have identified CAMK2D-associated transcript 1 (C2dat1) and 2 (C2dat2) (C2dat1-2) as two novel IR-induced lncRNAs that specifically regulated the expression of CaMKIIδ in rat and mouse models of focal cerebral IR. In our pilot study, C2dat1-2 mRNAs were upregulated in a time-dependent manner in mouse cortical penumbra after focal ischemic brain injury, which was accompanied by increased expression of CaMKIIδ at transcript and protein levels. The expression patterns of C2dat1-2 and CAMK2D were confirmed in mouse Neuro-2a cells in response to in vitro ischemia (oxygen-glucose deprivation/re-oxygenation, OGD/R). Knockdown of C2dat1 resulted in a significant blockade of CaMKIIδ expression, and potentiated OGD/R-induced cell death. Mechanistically, reduced CaMKIIδ expression upon silencing C2dat1 inhibited OGD/R-induced activation of the NF-κB signaling pathway. Further analysis showed that the downregulation of IKKα and further inhibition of IκBα degradation accounted for the inhibition of the NF-κB signaling activity by depleting C2dat1-2. Thus, C2dat1 appears to promote neuronal survival through regulating the NF-κB signaling pathway. Therefore, lncRNAs may be potential targets for therapeutic intervention of ischemia brain injury. Based on these preliminary findings, we hypothesize that C2dat1-2 are novel IR-induced lncRNAs that regulate the expression of CaMKIIδ to promote neuronal survival through the activation of the NF-κB signaling pathway. The primary goal of this application is to determine the function and signaling mechanisms of C2dat1-2, as well as the associated CaMKIIδ in IR- induced neuronal injury. The long-term goal is to gain more insights into the molecular bases of IR-associated biological processes and to identify novel therapeutic targets that confer neuroprotection during IR. Two specific aims are proposed: Specific Aim 1. Define the role of C2dat1-2 as novel ischemia-induced lncRNAs that promote neuronal survival by modulating CaMKIIδ expression in mouse primary neuronal cultures. Specific Aim 2. Determine if knockdown of C2dat1-2 potentiate IR-induced cell death in mouse model of focal cerebral ischemia and if the effects are mediated through down-regulation of CaMKIIδ.

描述（由适用提供）：局灶性脑缺血（缺血性中风）是美国残疾的最常见原因，也是美国第四主要死亡原因。尽管努力开发疾病的药物和手术治疗，但组织纤溶酶原激活剂（TPA）目前是唯一有效的治疗方法。更好地了解病理过程以及新靶标和疗法的发现将大大推动这一领域。关于脑缺血的LNCRNA的研究很少。 Through an lncRNA array analysis in a rat model of focal cerebral ischemia/reperfusion (IR), we have identified CAMK2D-associated transcript 1 (C2dat1) and 2 (C2dat2) (C2dat1-2) as two novel IR-induced lncRNAs that specifically regulated the expression of CaMKIIδ in rat and mouse models of focal cerebral IR.在我们的初步研究中，局灶性缺血性脑损伤后，在小鼠皮质阴茎中以时间依赖的方式上调C2DAT1-2 mRNA，这是通过在转录和蛋白质水平上增加Camkiiδ的表达来实现的。在小鼠神经2A细胞中证实了C2DAT1-2和CAMK2D的表达模式，以响应体外缺血（氧葡萄糖剥夺/重新氧化，OGD/R）。 C2DAT1的敲低导致CAMKIUδ表达的显着封锁，并可能导致OGD/R诱导的细胞死亡。从机械上讲，沉默C2DAT1时的CaMKIIδ表达降低，抑制了OGD/R诱导的NF-κB信号传导途径的激活。进一步的分析表明，IKKα的下调和IκBα降解的进一步抑制是通过消耗C2DAT1-2来抑制NF-κB信号传导活性的。这是C2DAT1通过调节NF-κB信号通路促进神经元存活。因此，LNCRNA可能是缺血脑损伤热干预的潜在靶标。基于这些初步发现，我们假设C2DAT1-2是新型IR诱导的LNCRNA，可以通过NF-κB信号传导途径激活CaMKIIδ的表达来促进神经元存活。该应用的主要目标是确定C2DAT1-2的功能和信号传导机制，以及IR诱导的神经元损伤中相关的CAMKIIδ。长期的目标是获得对IR相关生物学过程的分子碱基的更多见解，并确定在IR期间赋予神经保护作用的新型治疗靶标。提出了两个具体的目的：具体目标1。定义C2DAT1-2作为新型缺血诱导的LNCRNA的作用，通过调节小鼠原发性神经元培养物中的CAMKIIδ表达来促进神经元存活。具体目的2。确定局灶性脑缺血小鼠模型中C2DAT1-2潜在IR诱导的细胞死亡的敲低以及是否通过CAMKIIΔ下调介导的效果。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Protein Kinase D: A Potential Therapeutic Target in Prostate Cancer.

蛋白激酶 D：前列腺癌的潜在治疗靶点。

DOI：
发表时间：
2017
期刊：
Molecular and cellular pharmacology
影响因子：
0
作者：
Roy,Adhiraj;Wang,QJane
通讯作者：
Wang,QJane

Protein kinase D2 confers neuroprotection by promoting AKT and CREB activation in ischemic stroke.

DOI：
10.1016/j.nbd.2023.106305
发表时间：
2023-10-15
期刊：
NEUROBIOLOGY OF DISEASE
影响因子：
6.1
作者：
Connelly, Jaclyn A.;Zhang, Xuejing;Chen, Yuzhou;Chao, Yapeng;Shi, Yejie;Jacob, Tija C.;Wang, Q. Jane
通讯作者：
Wang, Q. Jane