Anti-Oxidant Promoting Cytokine LIF Enhances Neural Cell Survival During Ischemia

抗氧化促进细胞因子 LIF 增强缺血期间神经细胞的存活

• 批准号: 9268246
• 负责人: Keith R Pennypacker
• 金额: $ 32.7万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9268246
• 关键词: Address; Age; Anti-Inflammatory Agents; Anti-inflammatory; Antioxidants; Behavioral; Binding; Cell Survival; Cells; Clinic; Clinical; Data; Dose; Elderly; Electrophoretic Mobility Shift Assay; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; FDA approved; Female; Gene Expression; Health; Human; Immunohistochemistry; Infarction; Injury; Ischemia; Knockout Mice; LIF gene; Laboratories; Measures; Middle Cerebral Artery Occlusion; Modeling; Motor Skills; Neurologic; Neurons; Occlusion injury; Oligodendroglia; Performance; Protein Isoforms; Protein Kinase; Proteins; Public Health; Rattus; Records; Recovery; Recovery of Function; Regimen; Reperfusion Therapy; Rodent; Safety; Signal Transduction; Small Interfering RNA; Stroke; Superoxide Dismutase; Techniques; Therapeutic; Tissues; Translating; Treatment Protocols; activating transcription factor; base; combat; cytokine; functional outcomes; gray matter; improved; individualized medicine; interest; knock-down; male; pre-clinical; promoter; research study; response; sex; transcription factor; treatment strategy; white matter

 DESCRIPTION (provided by applicant): Leukemia inhibitory factor (LIF) is an anti-inflammatory cytokine that activates the expression of endogenous antioxidants in neural cells. Our laboratory has been examining the efficacy of LIF in the rodent stroke model of middle cerebral artery occlusion (MCAO), which reflects the most commonly occluded vessel in humans. We have found that LIF administered at 6 h after MCAO improves anatomical and functional outcomes in rats. Of interest, this treatment reflects a 33% extension of the therapeutic window when extrapolated to the current clinical stroke window for therapy. LIF increased the expression of antioxidant proteins via Akt in cultured neurons and oligodendrocytes. Aim 1: Is the efficacy of LIF dependent on SOD? We have found total superoxide dismutase (SOD) levels are significantly elevated and protein levels of one isoform, SOD3, are increased several-fold at 3 days following MCAO. In this Aim, we will determine if specific SOD subtype(s) are responsible for the LIF-induced effects and determine cell-specific expression of these SOD isoforms. MCAO studies will first use knockout mice to examine the SOD subtypes and these results will then be confirmed in rats. Aim 2: Does this LIF dosing regimen translate into long-term behavioral recovery? This Aim will determine whether this treatment regimen has lasting effects. These data will be used to devise a treatment regimen to extend the short-term efficacy to long-term histological and functional recovery at 90 days post-MCAO in rats. Aim 3: Which signals are transduced following LIF activation? The efficacy of LIF is dependent on the protein kinase Akt. From analysis of antioxidant promoters, the transcription factor MZF-1 is the putative transcription factor that activates their gene expression via Akt. We will use immunohistochemistry and gel shift assays to quantify their expression and promoter binding, respectively, in the infarct regions of rats treated with LIF after MCAO. siRNA techniques with neuronal cultures will be employed to knock down expression of this factor to validate previous findings. Aim 4: Will LIF administration be effective in elderly rats after MCAO? In this Aim, we will treat elderly male and female rats in short-term studies. These studies will b used to determine if there are any age- or sex- dependent differences in LIF efficacy after MCAO. Both histological and functional recovery will be examined. This proposal is aimed to further elucidate the mechanism(s) of LIF treatment and optimize the treatment regimen for long-term efficacy. While treatment with exogenous antioxidants has failed, we believe that the systemic activation of endogenous antioxidants is a better approach and could have substantial implications for treating clinical stroke.

 描述（由申请人提供）：白血病抑制因子（LIF）是一种抗炎细胞因子，可激活神经细胞中内源性抗氧化剂的表达。我们的实验室一直在研究LIF在啮齿动物大脑中动脉闭塞（MCAO）中风模型中的疗效，这反映了人类最常见的闭塞血管。我们已经发现，在MCAO后6小时给予LIF改善了大鼠的解剖和功能结果。值得注意的是，当外推至当前临床卒中治疗窗口时，该治疗反映了治疗窗口的33%扩展。LIF通过Akt增加神经元和少突胶质细胞抗氧化蛋白的表达。目的1：LIF的疗效是否依赖于SOD？我们已经发现，总超氧化物歧化酶（SOD）水平显着升高和蛋白质水平的一个亚型，SOD 3，增加了几倍，在3天后MCAO。在这个目标中，我们将确定是否特定的SOD亚型负责LIF诱导的效应，并确定这些SOD亚型的细胞特异性表达。MCAO研究将首先使用基因敲除小鼠来检查SOD亚型，然后在大鼠中证实这些结果。目的2：这种LIF给药方案是否能转化为长期的行为恢复？这一目标将决定这种治疗方案是否具有持久的效果。这些数据将用于设计治疗方案，以将短期疗效扩展至大鼠MCAO后90天的长期组织学和功能恢复。目的3：LIF激活后转导哪些信号？LIF的疗效依赖于蛋白激酶Akt。从抗氧化剂启动子的分析，转录因子MZF-1是通过Akt激活其基因表达的假定转录因子。我们将使用免疫组织化学和凝胶迁移分析，以量化其表达和启动子结合，分别在梗死区的LIF治疗后MCAO的大鼠。将采用神经元培养物的siRNA技术来敲低该因子的表达以验证先前的发现。目的4：LIF给药对MCAO后的老年大鼠有效吗？ 为此，我们将在短期研究中对老年雄性和雌性大鼠进行治疗。这些研究将B确定MCAO后LIF疗效是否存在任何年龄或性别依赖性差异。将检查组织学和功能恢复。该提案旨在进一步阐明LIF治疗的机制并优化治疗方案以获得长期疗效。虽然外源性抗氧化剂治疗失败，我们认为，内源性抗氧化剂的全身激活是一个更好的方法，并可能有实质性的影响，治疗临床中风。