Novel mechanism of neural and muscular degeneration

神经和肌肉退化的新机制

基本信息

批准号：
10624824
负责人：
Xin Jie Chen
金额：
$ 42.51万
依托单位：
UPSTATE MEDICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10624824
关键词：
Address Adenine Nucleotide Translocase Affect Aging Amyotrophic Lateral Sclerosis Animals Area Autophagocytosis Behavioral Bioenergetics Cause of Death Cell Death Cell Death Induction Cells Central Nervous System Charge Child Chronic progressive external ophthalmoplegia Clinical Cytoprotection Cytosol Data Defect Degenerative Disorder Dementia Disease Equilibrium Experimental Models FRAP1 gene Frontotemporal Dementia Functional disorder Future Gene Expression Profile Genes Genetic Transcription Goals Heart Histologic Human Impairment Inner mitochondrial membrane Intervention Knock-in Mouse Late-Onset Disorder Lateral Maintenance Membrane Missense Mutation Mitochondria Mitochondrial DNA Mitochondrial Diseases Mitochondrial Proteins Modeling Molecular Mus Muscle Muscle Weakness Mutation Myopathy Names Neuromuscular Diseases Neurons Nuclear Nucleotides Oxidative Phosphorylation Oxidative Phosphorylation Deficiency Oxidative Stress Parkinson Disease Pathogenicity Pathology Pathway interactions Peripheral Nervous System Diseases Phenotype Play Protein Import Protein Isoforms Proteins Repression Research Respiration Role SLC25A4 gene Seizures Skeletal Muscle Spastic Paraplegia Spinocerebellar Ataxias Stress Suppressor-Effector T-Lymphocytes Surface Syndrome System Testing Ubiquitination Validation Variant Yeasts aged cerebral atrophy dominant genetic mutation hearing impairment insight mitochondrial dysfunction mouse model muscle form mutant neural neuromechanism neuromuscular novel posttranscriptional preference psychiatric symptom receptor success tool translocase ubiquitin-protein ligase young adult

项目摘要

Ant1 is the muscle/heart/central nervous system (CNS) isoform of adenine nucleotide translocase that is primarily involved in ATP/ADP exchange across the inner mitochondrial membrane (IMM). An increasing number of missense mutations in Ant1 are found to cause dominant diseases that affect skeletal muscle and the central nervous system. These diseases are commonly manifested by fractional mtDNA deletions and mild bioenergetic defects. The mechanism of neuromuscular damage in the diseases is poorly understood. Interestingly, our recent studies in yeast and cultured human cells suggested that the mutant Ant1 is misfolded. This leads to cell death by a novel mechanism that we named mitochondrial Precursor Overaccumulation Stress (mPOS). mPOS is characterized by the toxic accumulation and aggregation of un-imported mitochondrial preproteins in the cytosol. These findings led to the central hypothesis that the mutant Ant1 primarily affects mitochondrial protein import. This results in mPOS in the cytosol, which plays an important role in inducing neural and muscular degeneration. Fractional mtDNA deletions occur independent of nucleotide transport activity, likely as a secondary damage collateral to reduced mitochondrial protein import. In this application, we propose to directly test this hypothesis in mouse models. We successfully generated knock-in (KI) mouse lines expressing misfolded variants of Ant1. Preliminary studies indicated that these mice develop phenotypes consistent with neural and muscular degeneration. In Specific Aim 1, we will use these unique experimental models to test the hypothesis that misfolded Ant1 induces neural and muscular degeneration and mtDNA instability independent of nucleotide transport. In Specific Aim 2, we will use various experimental tools that we developed in yeast, cultured human cells and the Ant1-KI mice to test the hypothesis that the misfolded Ant1 (or Aac2 in yeast) causes structural and functional damage to the mitochondrial protein import machinery and induces mPOS in the cytosol. In Specific Aim 3, we will determine the mechanisms that protect cells against Ant1-induced protein import stress and mPOS. Success of the project will establish a mouse model of protein import stress associated with mPOS. Particularly, validation of the mPOS model would help reconciling the mitochondrial and proteostatic pathways in many neural and muscular degenerative diseases. Finally, the results could have important implications for the understanding and therapy of Ant1-induced diseases, as well as many other clinical conditions that directly or indirectly affect mitochondrial protein import.

ANT1是腺嘌呤核苷酸易位酶的肌肉/心脏/中枢神经系统（CNS）同工型主要参与跨线粒体膜（IMM）的ATP/ADP交换。越来越多发现ANT1中的错义突变数量会导致影响骨骼肌肉和的主要疾病中枢神经系统。这些疾病通常由分数mtDNA缺失和轻度表现生物能缺陷。疾病中神经肌肉损伤的机制知之甚少。有趣的是，我们最近在酵母和培养的人类细胞的研究表明，突变体ANT1被错误折叠。这通过一种新的机制导致细胞死亡，我们将其命名为线粒体前体过度累积压力（MPO）。 MPO的特征在于未提示的有毒积累和聚集细胞质中的线粒体前蛋白。这些发现导致了突变体ANT1的中心假设主要影响线粒体蛋白的进口。这会导致细胞质中的MPO，这起着重要的作用在诱导神经和肌肉变性中的作用。分数mtDNA缺失独立于核苷酸转运活性，可能是降低线粒体蛋白进口的继发损伤。在此应用程序中，我们建议在小鼠模型中直接检验该假设。我们成功生成表达ANT1的错误折叠变体的敲入（Ki）小鼠系。初步研究表明这些小鼠发展与神经和肌肉变性一致的表型。在特定目标1中，我们将使用这些独特的实验模型，以测试错误折叠的ANT1的假设可诱导神经和肌肉变性和mtDNA不稳定性独立于核苷酸转运。在特定目标2中，我们将使用各种我们在酵母，培养的人类细胞和ANT1-KI小鼠中开发的实验工具以测试假设错误折叠的ANT1（或酵母中的AAC2）对结构和功能损害造成线粒体蛋白进口机制并在细胞质中诱导MPO。在特定目标3中，我们将确定保护细胞免受ANT1诱导的蛋白进口应力和MPO的机制。成功的成功项目将建立与MPO相关的蛋白质进口应力的小鼠模型。特别是验证 MPO模型将有助于调解许多神经和肌肉退化性疾病。最后，结果可能对理解具有重要意义和ANT1诱导疾病的治疗以及许多直接或间接影响的其他临床状况线粒体蛋白导入。