Regulations of organellar Zn2+ homeostasis and dynamics by TRPML1 in neurons

TRPML1 对神经元细胞器 Zn2 稳态和动力学的调节

基本信息

批准号：
10620676
负责人：
Yan Qin
金额：
$ 32.12万
依托单位：
UNIVERSITY OF DENVER (COLORADO SEMINARY)
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10620676
关键词：
Address Affect Aging Alzheimer&apos s Disease Axonal Transport Biological Biosensor Brain Brain region Cations Cell model Cells Chelating Agents Childhood Cytosol DNA Sequence Alteration Data Detection Disease Dominant-Negative Mutation Engineering Etiology Exhibits Fibroblasts Ganglioside Sialidase Deficiency Disease Genes Goals Health Hela Cells Hippocampus Homeostasis Human Impairment Ions Kinetics Knowledge Lead Lysosomal Storage Diseases Lysosomes Maps Measures Mediating Mental Depression Metals Mitochondria Mitochondrial Matrix Molecular Monitor Mutation Nerve Degeneration Neurites Neurodegenerative Disorders Neurons Organelles Parkinson Disease Pathogenesis Pathologic Patients Permeability Physiological Process Proteins RNA Interference Rattus Regulation Research Role Series Severities Signal Transduction Specificity Testing Time Vesicle Work improved inhibitor injured late endosome nanomolar nervous system disorder neurite growth neurodegenerative phenotype neuronal cell body new therapeutic target next generation novel postsynaptic presynaptic sensor small molecule tool vesicular release

项目摘要

The importance of labile, unbound Zn2+ in normal brain function has been evidenced by the association of abnormal cellular Zn2+ distributions with a series of neurological diseases such as depression, Alzheimer's disease (AD), aging, Mucolipidosis type IV disease (MLIV), and Parkinson’s diseases. MLIV is a lysosomal storage disease with neurodegenerative phenotype developed in childhood. The genetic mutations causing MLIV have been identified in the gene TRPML1, which encodes a lysosomal channel permeable to cations such as Ca2+and Zn2+. Loss of TRPML1 function resulted in elevated lysosomal Zn2+ in the fibroblasts derived from MLIV patients. However, neither the molecular mechanisms nor the biological impacts of such Zn2+ dysregulations are understood. Our preliminary studies devised a novel sensor GZnP3 with unprecedented sensitivity in the nanomolar range and provided the first profound evidence that TRPML1 can mobilize Zn2+ from lysosomes and late endosomes to the cytosol in neurons. Such endolysosomal Zn2+ release is unique in neurons and yields much greater Zn2+ signals in neurites than in the soma. Furthermore, we revealed that nanomolar Zn2+ can reduce the axonal transport in neurons. Based on the preliminary evidence, we hypothesize that impaired Zn2+ release from the vesicular organelles can cause neurodegeneration and contribute to the pathogenesis of MLIV. We will utilize the MLIV cell models along with our unique genetically targeted probes to test our hypothesis and address three specific aims: (1) We will quantify and compare the Zn2+ concentrations among various subcellular compartments in normal and MLIV cell models that are established in human fibroblasts and rat hippocampal neurons; (2) We will utilize our sensitive probes to investigate the correlation between impaired endolysosomal Zn2+ release with MLIV disease and determine the transport mechanisms that concentrate high pools of Zn2+ into endolysosomal vesicles in neurons; (3) We will examine our hypothesis that TRPML1 regulates neuronal health and function by mediating Zn2+ release from lysosomes to the cytosol: the released Zn2+ signals can regulate axonal transport and reduced lysosomal Zn2+ can recover the autophagic function of lysosomes. Collectively, the proposed research will provide ultra sensitive tools for monitoring cellular Zn2+ dynamics, develop a better understanding about the changes in organellar Zn2+ distributions and dynamics in MLIV, characterize the correlation between endolysosomal Zn2+ release and MLIV, as well as reveal the impacts of TRPML1-mediated Zn2+ dynamics on neuronal function. All of the above will significantly expand our knowledge about the pathological mechanisms of MLIV disease.

不稳定的、未结合的 Zn2+ 在正常大脑功能中的重要性已由以下因素的关联证明：细胞内Zn2+分布异常与抑郁症、阿尔茨海默病等一系列神经系统疾病疾病（AD）、衰老、粘脂沉积症 IV 型疾病（MLIV）和帕金森病是一种溶酶体疾病。儿童时期出现的具有神经退行性表型的储存病是由基因突变引起的。 MLIV 已在 TRPML1 基因中被发现，该基因编码可渗透阳离子的溶酶体通道如 Ca2+ 和 Zn2+。TRPML1 功能的丧失导致源自成纤维细胞的溶酶体 Zn2+ 升高。然而，对于 MLIV 患者，此类 Zn2+ 的分子机制和生物学影响都没有。我们的初步研究设计了一种具有前所未有的新型传感器 GZnP3。纳摩尔范围内的灵敏度，并提供了第一个深刻的证据，证明 TRPML1 可以从这种内溶酶体 Zn2+ 释放在神经元中是独特的。并且在神经突中产生比在体细胞中更大的 Zn2+ 信号。此外，我们还发现了纳摩尔。根据初步证据，我们发现 Zn2+ 可以减少神经元的轴突运输。囊泡细胞器释放的 Zn2+ 受损可导致神经变性并导致我们将利用 MLIV 细胞模型以及我们独特的基因靶向探针来研究 MLIV 的发病机制。检验我们的假设并实现三个具体目标：(1) 我们将量化并比较 Zn2+ 浓度在人类建立的正常和 MLIV 细胞模型中的各种亚细胞区室中 (2) 我们将利用我们的敏感探针来研究相关性内溶酶体 Zn2+ 释放受损与 MLIV 疾病之间的关系，并确定转运机制将大量 Zn2+ 浓缩到神经元的内溶酶体囊泡中；(3) 我们将检验我们的假设： TRPML1 通过介导 Zn2+ 从溶酶体释放到细胞质来调节神经元健康和功能：释放的 Zn2+ 信号可以调节轴突运输，溶酶体 Zn2+ 的减少可以恢复自噬总的来说，拟议的研究将为监测细胞提供超灵敏的工具。 Zn2+ 动力学，更好地了解细胞器 Zn2+ 分布和动力学的变化在 MLIV 中，表征了内溶酶体 Zn2+ 释放与 MLIV 之间的相关性，并揭示了 TRPML1 介导的 Zn2+ 动力学对神经功能的影响将显着扩展我们的研究。了解 MLIV 疾病的病理机制。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Neuronal signalling of zinc: from detection and modulation to function.

锌的神经元信号传导：从检测和调节到功能。

DOI：
发表时间：
2022-09
期刊：
影响因子：
5.8
作者：
Zhang, Chen;Dischler, Anna;Glover, Kaitlyn;Qin, Yan
通讯作者：
Qin, Yan

Spontaneous, synchronous zinc spikes oscillate with neural excitability and calcium spikes in primary hippocampal neuron culture.

在原代海马神经元培养物中，自发、同步的锌尖峰随神经兴奋性和钙尖峰振荡。

DOI：
发表时间：
2021
期刊：
影响因子：
4.7
作者：
Zhang, Chen;Maslar, Drew;Minckley, Taylor F;LeJeune, Kate D;Qin, Yan
通讯作者：
Qin, Yan

Foldamers reveal and validate therapeutic targets associated with toxic α-synuclein self-assembly.

Foldamers 揭示并验证与有毒 α-突触核蛋白自组装相关的治疗靶点。

DOI：
发表时间：
2022-04-27
期刊：
影响因子：
16.6
作者：
Ahmed, Jemil;Fitch, Tessa C;Donnelly, Courtney M;Joseph, Johnson A;Ball, Tyler D;Bassil, Mikaela M;Son, Ahyun;Zhang, Chen;Ledreux, Aurélie;Horowitz, Scott;Qin, Yan;Paredes, Daniel;Kumar, Sunil
通讯作者：
Kumar, Sunil

Development and Characterization of a Red Fluorescent Protein-Based Sensor RZnP1 for the Detection of Cytosolic Zn2.

用于检测胞质 Zn2 的红色荧光蛋白传感器 RZnP1 的开发和表征。