MicroRNA lipid-nanoparticle based therapy targets neuroinflammation and ApoE dysregulation in Alzheimer’s disease

项目标题：基于微RNA脂质纳米颗粒的治疗针对神经炎症和载脂蛋白E失调 在阿尔茨海默病中 摘要 MicroRNAs(MiRNA)是一种小的非编码调控RNA，在健康和疾病中有很大的影响。 虽然先前的研究表明miRNA与阿尔茨海默病和相关痴呆(ADRD)有关，但他们 尚未用于成功的治疗策略。我们发现microRNA-223(miR-223)，a 位于X染色体上的miRNA富含在小胶质细胞/巨噬细胞中，调节着性别 通过靶向CCAAT增强子结合蛋白β(CEBPβ)的二态载脂蛋白E途径。CEBPβ是一个重要的 调节神经炎性反应，并作为转录因子介导载脂蛋白E的表达。 我们最近的数据显示，miR-223缺乏导致显著的、女性偏见的高血压。 CEBPβ、ApoE与巨噬细胞和衰老小鼠脑内炎症状态的升高 脑部受伤。因此，miR-223似乎调节性二态小胶质细胞/巨噬细胞依赖 随着年龄、损伤和/或脑细胞的激活，炎症和载脂蛋白E增加 阿尔茨海默病相关病理学。我们进一步证明了miR-223可以通过一种新的 MiRNA-脂质体给药系统。基于这些令人振奋的初步结果，这项提案的主要目标是 正在测试miR-223-CEBPβ通路在调节炎症触发的载脂蛋白E异常调节中的作用。 年龄、性别和阿尔茨海默病相关脑损伤事件的生物学背景；并测试 新型脂质体-miRNA递送方法作为一种潜在的治疗策略。 为了完成这些目标，我们将1)确认miR-223直接靶向CEBPβ-apoE途径 2)检测脂质体-miR-223-CEBPβ- 使用脑损伤小鼠模型的APOE通路和使用脂质体miRNA递送的靶向通路 系统在一种新的miR-223基因敲除小鼠中；3)在人细胞中评估miR-223-CEBPβ-apoE通路 与阿尔茨海默病的神经病理改变平行。这些研究将包括对 脑组织来自世界一流的肯塔基大学阿尔茨海默病中心生物库。 资助研究的完成将扩大我们对监管机制的理解 炎症/载脂蛋白E通路在性别、衰老和脑损伤的背景下，最终推进了对 阿尔茨海默氏症。填补这些知识空白可能会对公共卫生产生持久的影响。而且，我们的 以miR-223为靶点的基于miRNA的创新治疗策略为一种新药的有效性提供了证据 目标人群，特别是女性，她们容易患上与神经炎和载脂蛋白E相关的疾病 调节失调，包括阿尔茨海默病和其他脑损伤。

Project Title: MicroRNA lipid-nanoparticle based therapy targets neuroinflammation and ApoE dysregulation in Alzheimer’s disease Abstract MicroRNAs (miRNA) are small non-coding regulatory RNA that have large impacts in health and disease. While prior studies have implicated miRNA in Alzheimer’s disease and associated dementias (ADRD), they have not been used yet in a successful therapeutic strategy. We discovered that microRNA-223 (miR-223), a miRNA residing on the X-chromosome which is enriched in microglia/macrophage cells, regulates a sexually dimorphic ApoE pathway via targeting CCAAT-enhancer-binding protein beta (CEBPβ). CEBPβ is an important regulator of neuroinflammatory response and acts as a transcription factor mediating ApoE expression. Our recent data showed that deficiency of miR-223 resulted in a substantial, female-biased elevation of CEBPβ, ApoE, and a heightened inflammatory state in macrophages and in aged mouse brain following a brain injury. Thus, miR-223 appears to regulate a sexually dimorphic microglia/macrophages-dependent increase in inflammation and ApoE as brain cells become activated in parallel with age, injury, and/or Alzheimer’s disease-related pathology. We further demonstrated that miR-223 can be targeted using a novel miRNA-liposome delivery system. Based on these exciting preliminary results, the main goals of this proposal are to test the miR-223-CEBPβ pathway in regulating inflammation-triggered ApoE dysregulation in the biological context of age, sex, and Alzheimer’s disease-relevant brain injury event; and, to test the efficacy of a novel liposome-miRNA delivery method as a potential therapeutic strategy. To complete these objectives, we will 1) confirm that miR-223 directly targets the CEBPβ-ApoE pathway in myeloid cells, and test whether liposome-miR-223 delivery affects this pathway; 2) test the miR-223-CEBPβ- ApoE pathway using a brain injury mouse model and target the pathway using the liposome miRNA delivery system in a novel miR-223 knockout mouse; 3) evaluate the miR-223-CEBPβ-ApoE pathway in human cells in parallel with neuropathological changes of Alzheimer’s disease. These studies will include assessments of brain tissue from the world-class University of Kentucky Alzheimer’s Disease Center biobank. Completion of the funded studies will extend our understanding of mechanisms regulating inflammation/ApoE pathways in the context of sex, aging, and brain injury, ultimately advancing treatments for Alzheimer’s disease. Filling these knowledge gaps could have a lasting impact on public health. Moreover, our innovative miRNA-based therapeutic strategy targeting miR-223 provides a proof of efficacy for a new drug target, particularly in females, who are vulnerable for diseases associated with neuroinflammation and ApoE dysregulation, including Alzheimer’s disease and other brain injury.