UV damage stimulation by ETS transcription factors in skin cancer

ETS 转录因子对皮肤癌的紫外线损伤刺激

• 批准号: 9904652
• 负责人: John J Wyrick
• 金额: $ 19.19万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904652
• 关键词: Apoptosis; Automobile Driving; Binding; Binding Proteins; Binding Sites; Biochemistry; Bioinformatics; Cell Differentiation process; Cells; DNA; DNA Binding; DNA Damage; DNA Structure; DNA lesion; Data; ETS1 gene; Elements; Etiology; Exposure to; Family; Family member; Future; Genes; Genome; Genomic Segment; Human; Human Genome; In Vitro; Individual; Induced Mutation; Kinetics; Lesion; Malignant Neoplasms; Maps; Methods; Modeling; Molecular; Mutagenesis; Mutate; Mutation; Nucleotides; Oncogenes; Predisposing Factor; Predisposition; Promoter Regions; Proteins; Protocols documentation; Pyrimidine Dimers; Recurrence; Resolution; Ribosomal Proteins; Running; Site; Skin; Skin Cancer; Succinate Dehydrogenase; Techniques; Telomerase; Testing; Therapeutic; UV induced; Ultraviolet Rays; Variant; carcinogenesis; density; genome sequencing; human DNA sequencing; in vitro activity; in vitro testing; insight; interest; melanocyte; melanoma; migration; molecular dynamics; novel; promoter; repair enzyme; repaired; transcription factor; tumor; tumorigenesis; ultraviolet; ultraviolet damage; ultraviolet irradiation; whole genome

PROJECT SUMMARY Exposure to ultraviolet (UV) light is the principal etiological agent for melanoma and other skin cancers. UV light induces damage to the cellular DNA, primarily cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts (6-4PPs). Sequencing of DNA from human skin cancers have revealed high levels of UV- induced mutations that are heterogeneously distributed across the genome. It has recently been discovered that UV-induced mutations are particularly enriched at the DNA binding sites of transcription factors. While this has been attributed to less efficient repair at such binding sites, it is also possible that variations in initial damage formation due to transcription factor binding could stimulate mutation rates. To test this hypothesis, we have used our newly developed CPD-seq method to map CPD formation across the human genome immediately following UV irradiation. Our preliminary data indicate that initial UV damage formation is significantly elevated at many transcription factor binding sites (TFBS), particularly binding sites for the E26 transformation-specific (ETS) family of transcription factors (TFs). Importantly, stimulation of CPD formation by ETS binding correlates with significantly higher mutation rates in melanoma tumors, indicating that variations in initial DNA damage formation are an important contributor to the mutational 'landscape' in human cancers. The overall objective of this proposal is to elucidate the mechanism by which ETS binding stimulates UV damage formation and determine whether ETS-induced CPD 'hotspots' drive recurrent mutagenesis at individual binding sites in melanoma tumors. To investigate the mechanism by which ETS protein binding stimulates UV damage formation, we will characterize the effects of selected ETS transcription factors on UV damage formation and repair in vitro (Aim I). In parallel, we will use molecular dynamics simulations to model how DNA binding by different ETS transcription factors predisposes dipyrimidine sequences to form UV photoproducts. Finally, we will examine whether ETS TF binding inhibits repair of CPD lesions in vitro. In Aim II, we will develop the CPD-capture-seq method to map the formation and repair of CPD lesions with high sequencing depth and single nucleotide resolution at specific genomic regions of interest, including sites of recurrent promoter mutations in melanoma and ETS binding sites. In parallel, we will map ETS binding sites in human melanocytes using the ChIP-exo method. Comparison of CPD-capture-seq data with UV-induced mutations identified in human melanomas will allow us determine with high resolution whether ETS-induced CPD 'hotspots' and repair inhibition are associated with recurrent mutations in skin cancer. These data should provide new insights into the etiology of some of the most recurrent mutations in melanoma, which occur at ETS binding sites. ETS transcription factors are known oncogenes that regulate many genes involved in cell differentiation, migration, proliferation, and apoptosis; hence, recurrent mutations at ETS binding sites likely contribute to carcinogenesis in skin cancer and could be exploited in future therapeutics.

项目总结 暴露在紫外线下是黑色素瘤和其他皮肤癌的主要病因。UV 光诱导细胞DNA损伤，主要是环丁烷嘧啶二聚体(CPDS)和6-4 摄影产品(6-4pps)。人类皮肤癌的DNA测序显示出高水平的紫外线- 在整个基因组中异质性分布的诱变。它是最近被发现的 紫外线诱导的突变在转录因子的DNA结合部位特别丰富。虽然这件事 归因于这些结合位点的修复效率较低，也可能是初始 转录因子结合导致的损伤形成可能会刺激突变率。为了检验这一假设，我们 已经使用我们最新开发的CPD-SEQ方法在整个人类基因组中绘制了CPD形成图 在紫外线照射后立即进行。我们的初步数据表明，最初的紫外线伤害形成 在许多转录因子结合位点(TFBS)显著升高，特别是E26的结合位点 转换特异性(ETS)转录因子(TF)家族。重要的是，通过刺激CPD的形成 在黑色素瘤中，ETS结合与显著更高的突变率相关，这表明 最初的DNA损伤形成是人类癌症突变“版图”的重要因素。 这项建议的总体目标是阐明ETS结合刺激UV的机制 破坏形成并确定ETS诱导的CPD‘热点’是否在 黑色素瘤肿瘤中的个体结合位点。探讨ETS蛋白结合的机制 刺激紫外线损伤的形成，我们将表征选定的ETS转录因子对紫外线的影响 体外损伤形成和修复(目标I)。同时，我们将使用分子动力学模拟来模拟 不同ETS转录因子与DNA的结合如何使二嘧啶序列易于形成紫外线 摄影产品。最后，我们将在体外检测ETS-TF结合是否抑制CPD损伤的修复。在AIM 第二，我们将开发CPD-Capture-Seq方法来绘制CPD高密度病变的形成和修复 特定基因组感兴趣区域的测序深度和单核苷酸分辨率，包括 黑色素瘤和ETS结合部位反复出现的启动子突变。同时，我们将在 人黑素细胞使用芯片-exo方法。CPD-CAPTURE-SEQ数据与紫外线诱导数据的比较 在人类黑色素瘤中发现的突变将使我们能够高分辨率地确定ETS是否导致 CPD“热点”和修复抑制与皮肤癌的复发突变有关。这些数据应该 为黑色素瘤中一些最常见的突变的病因学提供新的见解，这些突变发生在 ETS结合位点。Ets转录因子是已知的癌基因，调节许多与细胞有关的基因 分化、迁移、增殖和凋亡；因此，ETS结合位点的反复突变可能 有助于皮肤癌的发生，并可能在未来的治疗中被利用。

项目成果

Risk of physical injuries in children and teens with ophthalmic diagnoses in the OptumLabs Data Warehouse.

OptumLabs 数据仓库中进行眼科诊断的儿童和青少年身体受伤的风险。

• DOI: 10.1016/j.jaapos.2021.07.007
• 发表时间: 2021
• 期刊: Journal of AAPOS : the official publication of the American Association for Pediatric Ophthalmology and Strabismus
• 作者: Pineles,StacyL;Repka,MichaelX;Yu,Fei;Velez,FedericoG;Doppee,Danielle;Perez,Claudia;Sim,Danielle;Coleman,AnneL
• 通讯作者: Coleman,AnneL