Mechanisms of PhIP-induced dopaminergic neurotoxicity

PhIP 诱导多巴胺能神经毒性的机制

基本信息

批准号：
10595271
负责人：
Jason R Cannon
金额：
$ 156.34万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-15 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10595271
关键词：
2-Amino-1-Methyl-6-Phenylimidazo[4,5-b]pyridine Address Animal Model Autophagocytosis Biochemical Biological Models Brain Cell model Cells DNA Adduction DNA Adducts DNA Damage Data Data Set Disease Disease model Domestic Fowls Dopamine Environment Exhibits Fishes Genes Genomic DNA Grant Histological Techniques Human Impairment Link Literature Meat Mediating Mitochondria Mitochondrial DNA Mitochondrial Proteins Modeling Molecular Monophenol Monooxygenase Motor Nematoda Nerve Degeneration Neurons Nuclear Outcome Parkinson Disease Pathogenesis Pathologic Pathology Pathway interactions Prevalence Prevention Process Proteins Publications Rattus Research Rodent Model Role Testing Therapeutic Toxic effect Toxicology Translational Research Viral Viral Vector adduct brain metabolism cooking dietary heterocyclic aromatic amines in vivo insight metabolomics mitochondrial dysfunction nervous system disorder neuromelanin neuron loss neurotoxic neurotoxicity novel oxidative damage protein aggregation pyridine toxicant

项目摘要

Dopamine (DA)-ergic neurodegeneration is a pathological hallmark of Parkinson’s disease (PD) that produces the cardinal motor features. Major gaps in the literature remain on if and how common dietary exposures may contribute to pathogenesis. This proposal aims to address these gaps through highly mechanistic studies of neurotoxicity from dietary toxicants known as heterocyclic aromatic amines (HAAs). In the first cycle of R01ES025750, we made major advances demonstrating that HAAs produce selective DAergic neurotoxicity in cellular, nematode, and rodent model systems. We also identified HAA-induced oxidative damage, protein aggregation, autophagy disruption, and DNA adduct formation as key biochemical and molecular outcomes that are of critical importance to PD. Within this dataset, we have made overarching mechanistic advances that set the stage for a mechanism-of-action-focused renewal. First, neuromelanin (NM) is critical to HAA intracellular accumulation and neurotoxicity. This finding points to selectivity because NM is formed in catecholaminergic neurons in humans, and a critical translational need for NM cell and animal models in the study of HAAs (NM is lacking in most PD models). Second, HAAs selectively target mitochondria, again pointing to possible selectivity because DAergic neurons are especially sensitive to mitochondrial toxicity. Based on these data and the literature, we will test the following mechanistic hypothesis: HAA-induced DAergic neurotoxicity is mediated through biochemical interactions between NM and mitochondrial dysfunction that produce a neurotoxic cascade. We will test this hypothesis through three aims. In Aim 1, we will determine if NM-forming rats exhibit heightened HAA-induced DAergic neurotoxicity. In novel, NM-forming rats, we will assess HAA accumulation, HAA brain metabolism, and neurotoxicity to establish PD relevance. In Aim 2, we will identify mitochondrial targets that mediate HAA-induced neurotoxicity. We will discover the role of mitochondrial DNA adducts of HAAs in mediating neurotoxicity by quantifying adducts formed in mitochondrial versus genomic DNA. Further, we will identify HAA bioactivation pathways that lead to mitochondrial and genomic DNA adduct formation. Finally, we will identify specific mitochondrial gene and protein impairments resulting from DNA damage. In Aim 3, we will demonstrate connections between NM, mitochondrial dysfunction, and protein aggregation. Using cell-free, cellular and animal model systems, we will determine the effects of NM on HAA-mediated perturbations of mitochondrial function, autophagy (especially mitophagy), and the propagation of PD-relevant protein aggregation using biochemical and histological techniques. Overall, elucidation of interactions between NM, mitophagy/autophagy, and protein aggregation as critical to HAA neurotoxic mechanism of action is expected to significantly advance understanding of HAA- induced neurotoxicity and, more broadly, environmentally induced DAergic neurotoxicity. These studies are expected to significantly advance understanding of PD etiopathogenesis.

多巴胺（DA） - 能神经变性是帕金森氏病（PD）的病理标志基本电机功能。文献中的主要差距仍然存在于IF以及饮食中的常见暴露方式上对发病机理的贡献。该建议旨在通过高度机械的研究来解决这些差距饮食中有毒物质的神经毒性称为杂环芳香胺（HAAS）。在第一个周期 R01ES025750，我们取得了重大进步，证明HAA在中产生了选择性的DAEROTOIC神经毒性细胞，线虫和啮齿动物模型系统。我们还确定了HAA诱导的氧化损伤，蛋白质聚集，自噬破坏和DNA加合物形成，作为关键的生化和分子结果对于PD至关重要。在此数据集中，我们已经取得了总体机理的进步为以行动机理为重点的续订奠定了基础。首先，Neuromelanin（NM）对于HAA至关重要细胞内积累和神经毒性。该发现指向选择性，因为NM在人类的儿茶酚胺能神经元，以及对NM细胞和动物模型的关键翻译需求 HAAS的研究（在大多数PD模型中缺乏NM）。其次，哈斯有选择地靶向线粒体，再次指出可能的选择性，因为Daergic神经元对线粒体毒性特别敏感。基于在这些数据和文献上，我们将测试以下机械假设：HAA诱导的DAERGIC 神经毒性是通过NM和线粒体功能障碍之间的生化相互作用来介导的。产生神经毒性级联反应。我们将通过三个目标检验这一假设。在AIM 1中，我们将确定是否 NM形成的大鼠暴露了HAA诱导的DAERROTOCICIS。在小说，形成NM的大鼠中，我们将评估HAA积累，HAA脑代谢和神经毒性，以建立PD相关性。在AIM 2中，我们将确定介导HAA诱导神经毒性的线粒体靶标。我们将发现 HAA的线粒体DNA在介导神经毒性中的线粒体DNA通过量化在中介导的神经毒性中。线粒体与基因组DNA。此外，我们将确定导致的HAA生物活化途径线粒体和基因组DNA加合物的形成。最后，我们将确定特定的线粒体基因和 DNA损伤引起的蛋白质障碍。在AIM 3中，我们将展示NM之间的联系线粒体功能障碍和蛋白质聚集。使用无细胞，细胞和动物模型系统，我们将确定NM对HAA介导的线粒体功能的扰动的影响（尤其是线粒体），以及使用生化和组织学的PD相关蛋白聚集的传播技术。总体而言，阐明NM，线粒体/自噬和蛋白质聚集之间的相互作用对HAA神经毒性作用机制至关重要，预计将显着提高对HAA-的理解诱导的神经毒性，更广泛地诱导了环境诱导的DAERTER毒性。这些研究是预计将显着提高人们对PD疗法发生的理解。