The role of alpha-synuclein in nucleolar DNA double-strand break repair and cellular senescence

α-突触核蛋白在核仁 DNA 双链断裂修复和细胞衰老中的作用

基本信息

批准号：
10677497
负责人：
Moriah Rebecca Arnold
金额：
$ 5.27万
依托单位：
OREGON HEALTH & SCIENCE 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：
2023-07-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10677497
关键词：
Affect Age Aging Alzheimer&apos s Disease Animals Antibodies Binding Biogenesis Biological Assay Biotinylation Brain Breeding Cell Aging Cell Line Cell Nucleolus Cell Nucleus Cell Proliferation Cell Survival Cells Central Nervous System Clinical Complex Cutaneous Melanoma Cytoplasmic Inclusion DNA DNA Double Strand Break DNA Repair Data Development Diagnosis Disease Double Strand Break Repair Electron Microscopy Environmental Risk Factor Epidemiology Eukaryotic Cell Genes Genetic Transcription Genome Stability Genomic Instability Goals Growth Histologic Human Human Cell Line Image Immunofluorescence Microscopy Impairment In Vitro Incidence Individual Invaded Knock-out Knockout Mice Label Lewy Bodies Lewy Body Dementia Lewy Body Disease Link Malignant Neoplasms Measures Melanogenesis Melanoma Cell Metastatic Melanoma Metastatic Neoplasm to Lymph Nodes Methodology Methods Modeling Molecular Movement Mus Mutate Mutation Neoplasm Metastasis Nerve Degeneration Neurodegenerative Disorders Neurons Nuclear Parkinson Disease Pathologic Pathway interactions Patients Persons Phenotype Play Presynaptic Terminals Primary Neoplasm Proteomics Publishing Regulation Ribosomal DNA Ribosomes Risk Role SNCAIP gene Skin Cancer Substantia nigra structure Techniques Testing Tissues Up-Regulation Variant Work alpha synuclein cell growth disorder risk endonuclease epidemiology study gain of function genetic risk factor in vivo insight loss of function melanoma melanomagenesis mouse model mutant new therapeutic target novel overexpression repair function repaired senescence tumor tumor growth

项目摘要

PROJECT SUMMARY Genomic stability is very closely linked to diseases associated with aging, like neurodegeneration and cancer. Interestingly, strong evidence suggests unexpected links between these two diseases, since epidemiological studies have found that Parkinson’s Disease (PD) patients are at an increased risk of developing melanoma. Furthermore, this relationship is bidirectional; individuals diagnosed with melanoma are at an increased risk of developing PD. Although these clinical associations are well-established, the cellular and molecular pathways linking these diseases are poorly understood. PD is the second most common neurodegenerative disease associated with a complex combination of genetic and environmental risk factors, which manifests into devastating movement and coordination deficits. Its hallmark pathological finding is the presence of aggregated forms of alpha-synuclein (αSyn) in the substantia nigra of patient brains. Recent studies have also found a previously unrecognized role for αSyn in melanomagenesis and metastasis, highlighting the role of αSyn in regions outside of the central nervous system. Several studies have shown that melanoma cells overexpress αSyn and that this is important for promoting cell proliferation and growth; however, the underlying role of αSyn within melanogenesis is unknown. We have previously demonstrated that αSyn is important in DNA double- strand break (DSB) repair, and my new studies show an important role for αSyn within the nucleolus. Furthermore, genomic instability within the nucleolus has been associated with several diseases linked to cellular aging and has been shown to induce cellular senescence. Therefore, based on my preliminary data and these published findings, I hypothesize that the upregulation of αSyn plays a critical role in cells to facilitate nucleolar DSB repair, limit senescence, and thus contribute to overall cell survival. This hypothesis will be tested through two aims: first, I will determine the role of αSyn in nucleolar DSB repair (Aim 1) and next, determine the role of αSyn in cellular senescence inhibition and melanoma growth and metastatic potential (Aim 2). I will accomplish these goals through both in vitro and in vivo methodology. Using a human melanoma cell line that highly expresses αSyn, SK-Mel28, I will investigate the localization and function of αSyn in the nucleolus using various imaging and proteomic techniques. I will knockout αSyn to investigate loss-of-function nucleolar phenotypes and whether cellular senescence inhibition is altered. I will then reintroduce not only wildtype αSyn, but also four neurodegeneration-associated variants. To test whether αSyn plays a role in melanoma growth and metastasis through nucleolar DSB and senescence inhibition in vivo, I will breed an αSyn KO spontaneous melanoma mouse model. Through histological examination I will determine whether αSyn loss-of-function impairs nucleolar DSB repair and senescence inhibition, delays primary tumor growth, and diminishes metastasis. Achieving these goals will have a profound impact on the aging field by introducing genomic instability and cellular senescence concepts to our understanding of the cross-talk between PD and melanoma.

项目概要基因组稳定性与衰老相关疾病（如神经退行性疾病和癌症）密切相关。表明，强有力的证据表明这两种疾病之间存在意想不到的联系，因为流行病学研究发现帕金森病（PD）患者患黑色素瘤的风险增加。此外，这种关系是双向的；被诊断患有黑色素瘤的人患黑色素瘤的风险增加。尽管这些临床关联已得到充分证实，但细胞和分子途径。 PD 是第二常见的神经退行性疾病，但人们对这些疾病之间的联系知之甚少。与遗传和环境风险因素的复杂组合有关，这表现为其标志性病理发现是存在聚集性运动和协调缺陷。最近的研究还发现患者大脑黑质中存在多种形式的α-突触核蛋白（αSyn）。先前未被认识到 αSyn 在黑色素瘤发生和转移中的作用，强调了 αSyn 在一些研究表明，黑色素瘤细胞过度表达。 αSyn，这对于促进细胞增殖和生长很重要；然而，αSyn 的潜在作用我们之前已经证明 αSyn 在 DNA 双链中很重要。链断裂 (DSB) 修复，我的新研究表明 αSyn 在核仁内发挥着重要作用。此外，核仁内的基因组不稳定性与多种与细胞相关的疾病有关。因此，根据我的初步数据和这些数据，它会导致细胞衰老。已发表的研究结果，我认为 αSyn 的上调在细胞中发挥着关键作用，以促进核仁 DSB 修复，限制衰老，从而有助于整体细胞存活。这一假设将得到检验。两个目标：首先，我将确定 αSyn 在核仁 DSB 修复中的作用（目标 1），接下来，确定 αSyn 的作用 αSyn 在细胞衰老抑制和黑色素瘤生长和转移潜力中的作用（目标 2）。这些目标通过体外和体内方法使用高度的人类黑色素瘤细胞系。表达αSyn，SK-Mel28，我将使用各种方法研究αSyn在核仁中的定位和功能我将敲除 αSyn 以研究功能丧失的核仁表型和然后我不仅会重新引入野生型 αSyn，还会重新引入四种 αSyn。测试 αSyn 是否在黑色素瘤生长和转移中发挥作用。通过核仁DSB和体内衰老抑制，我将培育出αSyn KO自发性黑色素瘤通过组织学检查，我将确定 αSyn 功能丧失是否会损害核仁。 DSB 修复和衰老抑制，延迟原发性肿瘤生长，并减少转移。通过引入基因组不稳定性和细胞衰老，目标将对衰老领域产生深远影响概念来帮助我们理解 PD 和黑色素瘤之间的交互作用。