Alternative splicing in arsenical skin carcinogenesis

砷皮肤癌发生中的选择性剪接

• 批准号: 9979035
• 负责人: J CHRISTOPHER STATES
• 金额: $ 23.4万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-14 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9979035
• 关键词: 3' Untranslated Regions; 5' Untranslated Regions; Affect; Alternative Splicing; Area; Arsenic; Arsenicals; Arsenites; Attention; Automobile Driving; Biogenesis; Biological; Biological Process; Cell Cycle Regulation; Cell physiology; Cells; Chronic; Chronic Disease; Complex; Country; DNA Repair; DNA damage checkpoint; Data; Disease; Etiology; Event; Exposure to; Expression Profiling; Gene Expression; Gene Expression Regulation; Genome Components; Global Change; Health; Human; Individual; Lead; Longitudinal Studies; Malignant - descriptor; Malignant Neoplasms; Mass Spectrum Analysis; Messenger RNA; MicroRNAs; Mitotic; Modeling; Oncogenic; Organ; Outcome; Outcome Study; Oxidative Stress; PLK1 gene; Paint; Pathway Analysis; Pathway interactions; Phenotype; Play; Post-Transcriptional Regulation; Process; Production; Protein Isoforms; Proteins; Proteome; Proteomics; RNA Splicing; Regulation; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Sampling; Shapes; Signal Transduction; Skin; Skin Cancer; Skin Carcinogenesis; Squamous cell carcinoma; System; Testing; Time; Transcript; arsenic-induced carcinogenesis; base; biological adaptation to stress; carcinogenesis; contaminated drinking water; differential expression; genome-wide; keratinocyte; programs; proteomic signature; public health relevance; response; sodium arsenite; toxicant; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; tumorigenesis; well water

Project Summary Chronic exposure to arsenic, most commonly through contaminated drinking water, plagues over 200 million individuals in over 70 countries including the USA. Arsenic is a multi-organ toxicant and chronic arsenic exposure causes several chronic diseases including cancer in multiple organs, skin cancer being the most common. While several mechanisms are postulated to be responsible for arsenic-induced carcinogenesis, a clear picture is yet to emerge. It is well known that chronic arsenic exposure radically changes the transcriptomic and proteomic signatures, but the underlying mechanism for such sweeping global changes are not yet clearly understood. Differential alternative splicing plays a role in carcinogenesis and may be at play in arsenic-induced carcinogenesis. We employed state-of-the-art RNA-Seq analysis in a well-established model of arsenic-induced skin cancer (HaCaT cells exposed continuously to 100 nM sodium arsenite for 32 weeks) in a longitudinal study to understand the global changes in splicing occurring during the transformation process. Our data indicate >600 differential alternative splicing events occurred at each of the time points studied (7, 19, 28 weeks of exposure). This differential splicing program, by dramatically changing the proteome, could be a key player in the arsenic-induced transformation of skin cells. The present proposal aims to examine the contribution of significant differential splicing events at each time point towards the process of carcinogenesis. We will specifically scrutinize if the significant splicing events predicted by our transcriptomic studies can be correlated with protein isoform expression profiles. Furthermore, we also aim to study if predicted significantly different spliced isoforms that take place in the 5' or 3' UTR of transcripts can be correlated to expression profile in the mature mRNA samples from exposed and unexposed cells. The outcomes from this study will further our understanding of how alternative splicing shapes the cellular events taking place before, during and after the time the HaCaT cells become malignant. In addition, our study will look to elaborate the mechanistic basis of how one change in an upstream regulatory alternative splicing factor can cause genome-wide synchronized alternative splicing changes by signal amplification through successive steps, leading to altered proteome and ultimately adverse health effects.

项目摘要 长期暴露于砷（通常是通过受污染的饮用水）瘟疫超过2亿 包括美国在内的70多个国家 /地区的个人。砷是一种多器官毒物和慢性砷 暴露会导致多种慢性疾病，包括多个器官中的癌症，皮肤癌是最大的 常见的。虽然假设几种机制是砷诱导的癌变的原因 清晰的图片尚未出现。众所周知，慢性砷暴露从根本上改变了 转录组和蛋白质组学特征，但是这种全球变化的基本机制 尚未清楚地理解。差分替代剪接在癌变中起作用，可能在 在砷引起的致癌作用中发挥作用。我们在 砷诱导的皮肤癌（HACAT细胞连续暴露于100 nm钠）的公认模型 在一项纵向研究中，阿森矿持续32周），以了解在 转换过程。我们的数据表明> 600个差分替代剪接事件发生在每个 研究时间点（7、19、28周暴露）。这个差异剪接程序，通过急剧改变 蛋白质组可能是砷诱导的皮肤细胞转化的关键参与者。目前的提议 旨在检查每个时间点的重要差拼接事件的贡献 致癌过程。如果我们的重大剪接事件由我们的预测，我们将特别仔细检查 转录组研究可以与蛋白质同工型表达谱相关。此外，我们还旨在 研究如果预测在5'或3'转录本中发生的显着不同的剪接同工型可以 与暴露和未暴露细胞的成熟mRNA样品中的表达谱相关。这 这项研究的结果将进一步了解我们对替代剪接如何塑造细胞事件的理解 发生在HACAT细胞之前，期间和之后发生恶性肿瘤。此外，我们的研究将 旨在详细说明在上游调节替代剪接因子中如何变化的机械基础 可以通过连续的信号扩增引起全基因组同步的替代剪接变化 步骤，导致蛋白质组改变，最终导致健康影响。