Proteomic Core D

蛋白质组核心D

• 批准号: 10496284
• 负责人: Ying Zhu
• 金额: $ 12.71万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10496284
• 关键词: Action Potentials; Address; Affect; Aging; Auditory; Auditory area; Biochemical; Biochemistry; Biological Markers; Biological Process; Cell physiology; Cells; Cellular Structures; Clinical; Cochlear nucleus; Code; Complex; Cytometry; Demyelinations; Detection; Disease; FDA approved; Functional disorder; Genetic; Immunohistochemistry; Individual; Inhibitory Synapse; Joints; Liquid substance; Mass Spectrum Analysis; Measurement; Measures; Mediating; Modeling; Molecular; Nature; Nerve Degeneration; Neurons; Outcome; Persons; Pharmaceutical Preparations; Phase; Pitch Perception; Post-Translational Protein Processing; Predisposition; Presbycusis; Process; Program Research Project Grants; Property; Proteins; Proteome; Proteomics; Public Health; RNA; Regulation; Resolution; Resources; Sensorineural Hearing Loss; Speech Intelligibility; Speed; Stimulus; Synapses; Synaptic plasticity; Techniques; Testing; Therapeutic; age related; aged; aging auditory system; aging population; auditory pathway; blind; design; excitotoxicity; functional plasticity; improved; innovation; mouse model; neural; neural network; neuron loss; neurosensory; neurotransmission; protein biomarkers; response; single cell proteins; single-cell RNA sequencing; sound; spiral ganglion; tool

Abstract Complex biological processes like auditory neural aging are composed of dynamic changes in individual neurons' biochemistry, with various aging process susceptibilities. Direct measures of a single auditory neuron (spiral ganglion neuron (SGN) and cochlear nuclei (CN) neuron subtypes) and their age-related alterations are needed to characterize cellular transitions, regulatory mechanisms, and the microenvironment's contribution to age- related hearing loss (ARHL). A critical unmet need is determining the biochemical makeup of healthy and aged SGNs with single-cell resolution and a depth of coverage extending to thousands of biomolecules. Proteins mediate the bulk of cellular function, and the correlation between RNA and protein abundance is often lacking, and the proteome largely determines cellular structure and function. We hypothesize that the aging SGN undergoes a profound proteomic dynamic that is neuron subtype-specific. The aims of Core D are: 1. To refine liquid chromatograph-mass spectrometry (LC-MS)-based proteomics platform for sensitive and high-throughput SGN proteomics. 2. To apply the improved platform to study aging-related proteome regulations for SGN and CN subtypes and at single-cell resolution. The proposed effort will result in a processing and analysis platform capable of direct and unbiased profiling of SGNs and CN neurons with a depth of coverage of >3000 proteins/cell, thus providing a capability for direct protein measurement analogous to single-cell RNA-seq. Core D serves as a resource to identify ARHL biomarkers for Projects 1-3 to test and perform functional analyses. Outcomes will produce unbiased and in- depth single-cell protein profiles of SGNs and CN to provide a complete picture and identify protein biomarkers associated with ARHL.

摘要 复杂的生物学过程，如听觉神经老化，是由单个神经元结构的动态变化组成的。 生物化学，具有各种老化过程的敏感性。直接测量单个听觉神经元（螺旋 神经节神经元（SGN）和耳蜗核（CN）神经元亚型）及其年龄相关的变化是必要的 表征细胞的转变，调节机制，以及微环境对年龄的贡献- 相关性听力损失（ARHL）一个关键的未满足的需求是确定健康和老年人的生化组成， SGN具有单细胞分辨率和覆盖深度，可覆盖数千个生物分子。蛋白 介导大部分细胞功能，并且RNA和蛋白质丰度之间的相关性通常缺乏， 蛋白质组在很大程度上决定了细胞的结构和功能。我们假设老化的SGN 经历了一个深刻的蛋白质组动态，是神经元亚型特异性。 核心D的目标是： 1.完善基于液相色谱-质谱（LC-MS）的蛋白质组学平台， 高通量SGN蛋白质组学 2.应用改进的平台研究SGN和CN亚型衰老相关蛋白质组调控， 单细胞分辨率。 拟议的努力将产生一个处理和分析平台，能够直接和公正地分析 SGN和CN神经元的覆盖深度>3000蛋白/细胞，从而提供了直接检测的能力。 类似于单细胞RNA-seq的蛋白质测量。核心D作为识别ARHL的资源 项目1-3的生物标志物进行测试和功能分析。结果将产生无偏见和- SGN和CN的深度单细胞蛋白质谱，以提供完整的图像并识别蛋白质生物标志物 与ARHL有关。