Exosomal miRNA in neuron to astroglial communication in the CNS

中枢神经系统神经元与星形胶质细胞通讯中的外泌体 miRNA

• 批准号: 10435455
• 负责人: Yongjie Yang
• 金额: $ 42.87万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10435455
• 关键词: Address; Adult; Affect; Astrocytes; Biochemical; Biological; Blood Vessels; Brain; Caliber; Cell Communication; Cells; Cellular biology; Central Nervous System Diseases; Coculture Techniques; Communication; Complement 1q; Cre driver; Data; Development; Endosomes; Exercise; Genetic; Genetic Transcription; Glioblastoma; Glutamate Transporter; Human; Image; Immune; In Situ; In Vitro; Injections; Knowledge; Label; Ligands; Malignant Neoplasms; Mediating; MicroRNAs; Modeling; Molecular; Molecular Biology; Monitor; Morphogenesis; Morphology; Motor Neurons; Mus; Nervous System Trauma; Neurodegenerative Disorders; Neuroglia; Neurons; Outcome; PHluorin; Pathologic; Pathway interactions; Physiology; Process; Publishing; Regulation; Role; Sampling; Signal Transduction; Spinal; Surface; Synapses; Synaptic Transmission; Testing; Vesicle; Virus; base; cell type; designer receptors exclusively activated by designer drugs; exosome; extracellular vesicles; fluorescence imaging; genome-wide; in vivo; insight; intercellular communication; mouse genetics; myelination; nervous system disorder; neurotransmitter release; notch protein; novel; receptor; synaptic function; transcriptome

Abstract Neuron to (astro)glia communication is essential for functional synaptic transmission and physiology in the CNS. Despite the important modulatory roles of astroglia in synapse function, molecular pathways that regulate the neuron-astroglia functional unit are largely undefined. Exosomes (50-150 nm in diameter), a major type of secreted extracellular vesicles (EVs), are derived from intraluminal vesicles (ILVs) in the early endosomal compartment and are released from cells during endosome maturation. EVs and exosomes secreted from various CNS cell types have emerged as a novel and important intercellular communication pathway in the CNS. In particular, miRNAs (miRs) are often found in exosomes to shuttle between cells for intercellular signaling. Intercellular transfers of miRs have been observed in CNS cells to regulate glutamate transporter function, promote myelination, and maintain brain vascular integrity. Exosomal signaling has also been implicated in pathological conditions of the CNS, including neurological injury, neurodegenerative diseases, and glioblastoma. Despite the strong rigor in prior studies to suggest the importance of the exosomal pathway in CNS cell communication, these studies are largely based on culture models or human CSF samples, exosome signaling in situ in the CNS remains essentially unexplored. In addition, fundamentally important cell biology aspects of this pathway, such as neuronal activity's influence, exosome internalization mechanisms, and downstream regulation in recipient CNS cells also remain unknown. This is particularly important to address as CNS cell types are highly distinct from cancer/immune cells where most of exosome knowledge is currently gained and exosome signaling mechanisms can be very cell-type heterogeneous. Based on our published study and additional preliminary results, we propose the following aims in this project: Aim 1: Determine the effect of neuronal activity on the subcellular localization of ILVs and neuronal exosome secretion; Aim 2: Dissect recognition pathways and entry mechanisms involved in astroglial internalization of neuronal exosomes; Aim 3: Investigate genetic regulation of neuronal exosomal miR-124 in astroglia; We have generated a large amount of preliminary data to support our rationales and to demonstrate feasibility for proposed aims. We will employ mouse genetics, molecular biology, virus injections, various imaging, and biochemical approaches to complete these aims. Outcomes from this project will present in vivo evidence to support a previously unrecognized mode of communication from neurons to glia in the CNS. It will also provide much-needed cell biological knowledge and insights for understanding exosome signaling in neuron to glia communication, especially about miR-124-3p's non-cell autonomous genetic regulation in astroglia following its internalization. As altered neuron to (astro)glia communication is clearly implicated in many neurological diseases, this knowledge and insights can significantly help understand how this pathway is involved in CNS diseases.

抽象的 神经元与（星形）神经胶质细胞的通讯对于功能性突触传递和生理学至关重要 中枢神经系统。尽管星形胶质细胞在突触功能中具有重要的调节作用，但其分子途径 神经元-星形胶质细胞功能单位的调节很大程度上是不确定的。外泌体（直径50-150 nm）是一种主要的 分泌性细胞外囊泡（EV）的类型，源自早期内体中的管腔内囊泡（ILV） 室并在内体成熟期间从细胞中释放。 EV 和外泌体 各种中枢神经系统细胞类型已成为中枢神经系统中一种新颖且重要的细胞间通讯途径。 特别是，miRNA (miR) 经常存在于外泌体中，在细胞之间穿梭以进行细胞间信号传导。 在中枢神经系统细胞中观察到 miR 的细胞间转移可调节谷氨酸转运蛋白功能， 促进髓鞘形成，维持脑血管完整性。外泌体信号传导也与 中枢神经系统的病理状况，包括神经损伤、神经退行性疾病和胶质母细胞瘤。 尽管先前的研究非常严格地表明外泌体途径在中枢神经系统细胞中的重要性 沟通，这些研究主要基于培养模型或人类脑脊液样本、外泌体信号传导 中枢神经系统中的原位基本上尚未被探索。此外，基本重要的细胞生物学方面 该通路，如神经元活动的影响、外泌体内化机制以及下游 受体中枢神经系统细胞的调节也仍然未知。作为 CNS 细胞，这一点尤为重要 类型与癌症/免疫细胞高度不同，目前大多数外泌体知识都是在癌症/免疫细胞中获得的，并且 外泌体信号传导机制可能存在很大的细胞类型异质性。 根据我们发表的研究和其他初步结果，我们提出以下目标： 项目：目标 1：确定神经元活动对 ILV 和神经元亚细胞定位的影响 外泌体分泌；目标 2：剖析星形胶质细胞的识别途径和进入机制 神经元外泌体的内化；目标 3：研究神经元外泌体 miR-124 的遗传调控 星形胶质细胞;我们已经生成了大量的初步数据来支持我们的理由并证明 拟议目标的可行性。我们将采用小鼠遗传学、分子生物学、病毒注射、各种成像、 和生化方法来完成这些目标。该项目的成果将提供体内证据 支持中枢神经系统中从神经元到神经胶质细胞的一种以前未被识别的通信模式。它还将提供 了解神经元到神经胶质细胞的外泌体信号传导急需的细胞生物学知识和见解 通讯，特别是关于 miR-124-3p 在星形胶质细胞中的非细胞自主遗传调控 内化。由于神经元与（星形）神经胶质细胞通讯的改变显然与许多神经系统疾病有关 疾病，这些知识和见解可以显着帮助理解该途径如何参与中枢神经系统 疾病。