MiR-193a-5p as a Regulator of Intersectin1-short and Clathrin-Mediated Endocytosis in Alzheimer's Disease.

MiR-193a-5p 作为阿尔茨海默病中 Intersectin1-short 和网格蛋白介导的内吞作用的调节剂。

• 批准号: 10480814
• 负责人: Sierra Smith
• 金额: $ 5.01万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-05 至 2023-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10480814
• 关键词: 3' Untranslated Regions; 3xTg-AD mouse; Abeta clearance; Address; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer’s disease biomarker; Amyloid beta-Protein; Astrocytes; Autopsy; Binding; Bioinformatics; Biological; Biological Assay; Biological Markers; Brain; Brain Pathology; Cause of Death; Cells; Cerebrospinal Fluid; Clathrin; Data; Dementia; Development; Diagnosis; Disease; Endocytosis; Functional disorder; Future; Gene Expression; Genetic Transcription; Genome; Hippocampus (Brain); Hormones; Human; Immunoblot Analysis; In Situ Hybridization; In Vitro; Injections; Knowledge; Lentivirus Vector; Link; Living Donors; Luciferases; Measures; Mediating; Messenger RNA; MicroRNAs; Microglia; Mus; Nerve Degeneration; Neurofibrillary Tangles; Neurologic; Neurons; Pathogenesis; Pathology; Process; Protein Isoforms; Proteins; Publishing; Regulation; Reporter; Reporting; Role; Sampling; Scaffolding Protein; Senile Plaques; Small RNA; Testing; Transferrin; Translations; United States; Wild Type Mouse; bioinformatics pipeline; brain cell; brain tissue; cell type; circulating microRNA; coated pit; cohort; differential expression; fluorescence imaging; frontal lobe; in vivo; inhibitor; interest; microRNA biomarkers; mind control; mouse model; novel therapeutic intervention; overexpression; protein expression; tool; uptake

PROJECT SUMMARY Alzheimer’s disease (AD) is the most common form of dementia and is characterized by the development of amyloid beta (Aβ) plaques and neurofibrillary tangles in the brain. Currently, the pathogenesis underlying AD is unclear and there are no reliable tools to detect early changes in the brain. Studies in the Saugstad lab revealed a specific set of microRNA (miRNA) biomarkers that are decreased in cerebrospinal fluid (CSF), and which can classify AD patients from neurologically normal controls. MiRNAs regulate post-transcriptional gene expression via complementary binding to sequences in mRNA that in turn regulate translation and expression of the resulting protein. However, there is a knowledge gap regarding which mRNAs are targeted and regulated by the AD CSF miRNAs and how these target proteins may contribute to AD pathophysiology. To address this gap, I developed a bioinformatics pipeline to predict mRNA targets of the AD CSF miRNAs, and identified Intersectin1-short (ITSN1-s) as a protein targeted by the AD CSF miRNA, miR-193a-5p. Immunoblot analysis of human post mortem hippocampus showed significant increases in ITSN1-s protein in AD relative to control. However, as ITSN1-s is expressed in multiple brain cell types, further studies that determine the specific cell type(s) with increased ITSN1-s expression in AD are needed to examine potential mechanisms underlying AD. Published studies report that ITSN-s is expressed in astrocytes and microglia, with low expression in neurons, and that ITSN1-s functions to reduce clathrin-mediated endocytosis (CME). Microglia have been shown to uptake fibrillar Aβ through a clathrin-mediated mechanism, and CME disruption in AD has been linked to reduced Aβ clearance in neurons. However, it is not known whether ITSN1-s in microglia contributes to this pathophysiology in AD. This proposal will assess the regulatory role of miR-193a-5p on ITSN1-s and its functional significance in CME and AD. In order to accomplish this, I propose the following aims: 1. establish the cellular expression of miR-193-a-5p and ITSN1-s in AD microglia and neurons; 2. investigate the interaction between miR-193a-5p and ITSN1-s mRNA and the effects on ITSN1-protein expression in vitro and in vivo; and 3. determine the role of miR-193a-5p levels on CME and Aβ uptake in AD microglia. Ultimately, these studies will provide a deeper understanding of ITSN1-s regulation in AD and how this may contribute to AD pathology. In addition, this framework can be used in future studies to evaluate the biological consequences of circulating miRNA biomarkers in AD.

项目概要 阿尔茨海默病 (AD) 是最常见的痴呆症，其特征是出现 目前，AD 的发病机制是大脑中的淀粉样蛋白 (Aβ) 斑块和神经原纤维缠结。 索格斯塔实验室的研究尚不清楚，也没有可靠的工具来检测大脑的早期变化。 揭示了一组特定的 microRNA (miRNA) 生物标志物，这些标志物在脑脊液 (CSF) 中减少，并且 它可以将 AD 患者与神经正常对照进行分类。miRNA 调节转录后基因。 通过与 mRNA 序列互补结合来表达，进而调节翻译和表达 然而，对于哪些 mRNA 是靶向的以及哪些 mRNA 存在知识差距。 AD CSF miRNA 的调节以及这些靶蛋白如何促进 AD 病理生理学。 为了解决这个差距，我开发了一个生物信息学管道来预测 AD CSF miRNA 的 mRNA 目标，并且 鉴定出 Intersectin1-short (ITSN1-s) 是 AD CSF miRNA miR-193a-5p 的靶向蛋白。 对人类死后海马体的分析显示，与 AD 相比，ITSN1-s 蛋白显着增加 然而，由于 ITSN1-s 在多种脑细胞类型中表达，需要进一步的研究来确定。 需要在 AD 中 ITSN1-s 表达增加的特定细胞类型来检查潜在机制 已发表的研究报告指出，ITSN-s 在星形胶质细胞和小胶质细胞中表达，且表达量较低。 ITSN1-s 可以减少网格蛋白介导的内吞作用 (CME)。 已被证明可以通过网格蛋白介导的机制摄取纤维状 Aβ，并且 AD 中的 CME 破坏已 与神经元中 Aβ 清除率降低有关，但目前尚不清楚小胶质细胞中是否存在 ITSN1-s。 该提案将评估 miR-193a-5p 对 AD 的病理生理学作用。 ITSN1-s 及其在 CME 和 AD 中的功能意义 为了实现这一目标，我提出以下建议。 目的： 1. 建立 AD 小胶质细胞和神经元中 miR-193-a-5p 和 ITSN1-s 的细胞表达； 研究 miR-193a-5p 和 ITSN1-s mRNA 之间的相互作用以及对 ITSN1-蛋白的影响 体外和体内表达；以及 3. 确定 miR-193a-5p 水平对 AD 中 CME 和 Aβ 摄取的作用 最终，这些研究将提供对 AD 中 ITSN1-s 调节及其如何调节的更深入的了解。 这可能有助于 AD 病理学。此外，该框架可用于未来的研究来评估 AD 病理学。 AD 中循环 miRNA 生物标志物的生物学后果。