Mechanisms regulating the early stages of UV-induced skin cancer

调节紫外线诱发皮肤癌早期阶段的机制

• 批准号: 9904163
• 负责人: John T. Seykora
• 金额: $ 36.23万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904163
• 关键词: Actinic keratosis; Address; Affect; American; Automobile Driving; Basic Science; Biological; Biological Assay; Biological Markers; Biopsy; Biopsy Specimen; Carcinogens; Carcinoma in Situ; Cells; Cessation of life; Chromatin; Clonal Expansion; Clone Cells; Comparative Genomic Analysis; Consumption; DNA Damage; DNA Repair; DNA Sequence Alteration; DNA analysis; DNA sequencing; Data; Development; Differentiation and Growth; Dominant-Negative Mutation; Engineering; Epidemic; Epidermis; Flow Cytometry; Frequencies; Genes; Genetic; Genetically Engineered Mouse; Genome; Genomic DNA; Goals; Growth; Harvest; Health Care Costs; Healthcare; Histologic; Human; Human Engineering; Image; Incidence; Induced Mutation; Lead; Lesion; Libraries; Link; Malignant Neoplasms; Modeling; Morbidity - disease rate; Mus; Mutate; Mutation; Mutation Analysis; Neoplasm Metastasis; Nuclear; Nuclear Pore; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Oncogenes; Patients; Population; Probability; Proteins; Public Health; Quality of life; RNA analysis; Reporting; Research; Role; Sampling; Signal Transduction; Skin; Skin Cancer; Skin Carcinogenesis; Squamous cell carcinoma; Strategic Planning; Subfamily lentivirinae; Surgeon; System; TP53 gene; Testing; Therapeutic; Time; Tissues; Treatment Cost; UV Radiation Exposure; UV carcinogenesis; UV induced; UV induced DNA damage; UV sensitive; Ultraviolet Rays; Xenograft Model; Xenograft procedure; aging population; cellular transduction; combat; cost effective; design; effective therapy; epidermis cell; exome sequencing; genome integrity; human model; improved; in vivo; insight; keratinocyte; laser capture microdissection; loss of function mutation; mortality; mouse model; mutant; notch protein; novel; novel strategies; premalignant; response; skin squamous cell carcinoma; skin xenograft; small hairpin RNA; transcription factor; transcriptome sequencing; two photon microscopy; ultraviolet irradiation

Project Summary: Cutaneous squamous cell carcinoma (cSCC) is the second most common form of cancer with approximately 700,000 cases annually in the US leading to 3,000 deaths. The incidence of cSCC will increase given an aging population with rising cumulative UV exposure. In response to this epidemic, the Surgeon General issued a `Call to Action' detailing a plan to address the growing epidemic of UV-induced skin cancers. One component of this strategic plan to decrease UV-induced skin cancer is to perform research that defines the mechanisms of UV-induced skin cancer. The transition from keratinocyte to cSCC by UV radiation involves formation of precancerous lesions, called actinic keratoses (AKs) and carcinoma in situ (SCCIS). AKs and SCCIS are hypothesized to arise from UV-induced mutations in keratinocytes that lead to cell populations with aberrant growth and differentiation. To better understand how UV irradiation alters keratinocytes and promotes the early stages of skin cancer, we performed laser capture microdissection of SCCIS and adjacent UV-exposed epidermis, isolated genomic DNA and generated libraries for whole exome sequencing. This novel approach permits a precise comparative genomic analysis of mutations in UV-exposed epidermis and SCCIS. Our exomic sequencing data demonstrate low frequency UV-signature loss-of-function mutations in Notch in 60% of epidermal samples and heterozygous clonal mutations in 40% of SCCIS samples. Nucleoporins (Nups) were also frequently mutated in 80% of epidermal libraries and 100% of SCCIS libraries. These data show that UV-signature mutations in Notch and Nups are more common than mutations in known oncogenes such as p53 or RAS. The goal of this proposal is to show how Notch and Nup mutations promote the early stages of UV-induced skin carcinogenesis leading to SCCIS. The biological impact of UV- irradiation on Notch-deficient clones will be studied using genetically engineered mice and engineered human keratinocytes in xenograft models. The biological significance of Nup mutations in UV-irradiated skin will be determined using mice with an epidermal deficiency in the Nup Elys, which links the nuclear pore complex to chromatin. In addition, human keratinocytes will be engineered to express mutant Nups and subjected to UV-irradiation in a xenograft model. The aims of this proposal will yield novel insights into the role of Notch and Nup mutations in skin cancer and will provide important insights into designing rational approaches to treat SCCIS.

项目摘要：皮肤鳞状细胞癌（CSCC）是第二常见的 在美国，每年约有70万例病例的癌症形式导致3,000例死亡。 鉴于人口老龄化，累积紫外线增加，CSCC的发病率将增加 接触。为了回应这种流行病，外科医生发布了一个“行动呼吁”，详细介绍了 计划解决紫外线诱导的皮肤癌的流行病。一个组成部分 减少紫外线诱发的皮肤癌的战略计划是进行研究，以定义 紫外线诱导的皮肤癌的机制。紫外线从角质形成细胞到CSCC的过渡 辐射涉及形成癌性病变，称为活化性角膜生成（AKS）和 原位癌（SCCIS）。 AKS和SCCI被认为是由紫外线引起的 角质形成细胞中导致细胞群体异常生长和分化的突变。 更好地了解紫外线辐射如何改变角质形成细胞并促进早期阶段 皮肤癌，我们进行了激光捕获SCCI和相邻紫外线暴露的显微解剖 表皮，分离的基因组DNA和生成用于整个外显子组测序的文库。这 新方法允许对暴露于紫外线的突变进行精确的比较基因组分析 表皮和SCCIS。我们的例外测序数据显示低频紫外线签名 60％的表皮样品和杂合克隆中的缺点突变 40％SCCIS样品中的突变。核孔（NUP）也经常在80％中突变 表皮库和100％的SCCIS库。这些数据表明紫外线签名 Notch和NUP中的突变比已知的肿瘤基因中的突变（例如 p53或RAS。该提议的目的是展示Notch和NUP突变如何促进 紫外线引起的皮肤致癌作用的早期阶段导致SCCIS。紫外线的生物学影响 将使用基因工程小鼠和 在异种移植模型中设计的人角质形成细胞。 NUP的生物学意义 使用表皮缺乏的小鼠确定紫外线辐照皮肤的突变 将核孔复合物与染色质联系起来的NUP Elys。另外，人类 角质形成细胞将被设计为表达突变体NUP，并在 异种移植模型。该提案的目的将产生对Notch和Notch作用的新见解。 皮肤癌中的NUP突变，将为设计理性提供重要的见解 治疗SCCI的方法。