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

• 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
• 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

 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（由申请人提供）： 局灶性脑缺血（缺血性中风）是美国最常见的致残原因，也是第四大死亡原因，尽管在开发该疾病的药物和手术治疗方法方面做出了努力，但组织纤溶酶原仍然存在。激活剂（tPA）是目前唯一有效的疗法，更好地了解病理过程以及新靶点和疗法的发现将显着推进该领域的发展。通过对局灶性脑缺血/再灌注 (IR) 大鼠模型的 lncRNA 阵列分析，我们发现 CAMK2D 相关转录本 1 (C2dat1) 和 2 (C2dat2) (C2dat1-2) 是两种新型 IR 诱导的 lncRNA，它们特异性调节在我们的初步研究中，C2dat1-2 mRNA 在局灶性脑 IR 的大鼠和小鼠模型中表达上调。局灶性缺血性脑损伤后小鼠皮质半暗带中的 C2dat1-2 和 CAMK2D 表达模式在体外得到证实，并伴随着 CaMKIIδ 转录物和蛋白水平的表达增加。缺血（缺氧/复氧，OGD/R） C2dat1 的敲低导致 CaMKIIδ 表达的显着阻断，以及从机制上讲，沉默 C2dat1 后 CaMKIIδ 表达减少会抑制 OGD/R 诱导的 NF-κB 信号通路激活，进一步分析表明 IKKα 的下调和 IκBα 降解的进一步抑制是这种抑制的原因。通过消耗 C2dat1-2 来抑制 NF-κB 信号传导活性 因此，C2dat1 似乎可以通过调节 NF-κB 来促进神经元存活。因此，lncRNA可能是缺血性脑损伤治疗干预的潜在靶标，基于这些初步发现，我们发现C2dat1-2是一种新型IR诱导的lncRNA，可通过激活调节CaMKIIδ的表达来促进神经元存活。本应用的主要目标是确定 C2dat1-2 以及 IR 诱导神经元中相关 CaMKIIδ 的功能和信号传导机制。长期目标是更深入地了解 IR 相关生物过程的分子基础，并确定在 IR 期间提供神经保护的新治疗靶点： 具体目标 1. 定义 C2dat1- 的作用。 2 作为新型缺血诱导的 lncRNA，通过调节小鼠原代神经元培养物中的 CaMKIIδ 表达来促进神经元存活。 具体目标 2. 确定 C2dat1-2 的敲低是否会增强。局灶性脑缺血小鼠模型中 IR 诱导的细胞死亡，以及该效应是否通过 CaMKIIδ 下调介导。

项目成果

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。

• 发表时间: 2017
• 期刊: Molecular and cellular pharmacology
• 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