Measuring nucleotide excision repair in human populations

测量人群中的核苷酸切除修复

基本信息

批准号：
10202603
负责人：
Laura Jane Niedernhofer
金额：
$ 42.58万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-21 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10202603
关键词：
Address Affect American Amish Autoradiography Basal cell carcinoma Biological Assay Biopsy Blood Blood Cells Cell Cycle Cell Line Cell Nucleus Cells Chemistry Chemotherapy-Oncologic Procedure Child Clinic Clinical DNA DNA Damage DNA Repair DNA Structure DNA biosynthesis DNA lesion Defect Deoxyuridine Dermal Detection Diagnosis Diagnostic Disease Dose Enrollment Environmental Carcinogens Environmental Exposure Equipment Excision Exposure to Failure Family Fibroblasts Flow Cytometry General Population Genes Genetic Polymorphism Genome Genomic Instability Goals Grain Head and Neck Cancer Health Health Care Costs Heterozygote Human Individual Inherited Knowledge Label Lesion Link Longevity Lung Malignant Neoplasms Malignant neoplasm of lung Maryland Measurement Measures Methods Microscopy Mutation Nerve Degeneration Neurodegenerative Disorders Nuclear Nucleosides Nucleotide Excision Repair Nucleotides Oligonucleotides Oncology Patients Peripheral Blood Mononuclear Cell Persons Population Population Study Proteins Radioactive Radiolabeled Reproducibility Residual state Resistance Risk S Phase Sampling Series Severities Site Skin Skin Cancer Skin Carcinoma Spouses Squamous Cell Squamous cell carcinoma Sunburn Sunlight Symptoms Testing Thymidine Time Tissues UV induced UV induced DNA damage Ultraviolet Rays United States National Institutes of Health Universities Xeroderma Pigmentosum analog cancer therapy clinical diagnostics cohort design disorder prevention disorder risk early onset environmental stressor gene product gene repair genetic pedigree high risk inter-individual variation lymphoblast medical schools melanoma minimally invasive nucleoside analog photolesion placental mammal premature programs repaired sample collection tool ultraviolet ultraviolet irradiation virtual

项目摘要

PROJECT SUMMARY Nucleotide excision repair (NER) is a DNA repair mechanism that recognizes and removes bulky, helix-distorting lesions from the nuclear genome. Key substrates for NER are lesions induced by ultraviolet (UV) radiation upon environmental exposure to sunlight and a subset of oxidative DNA lesions produced endogenously. This is dramatically illustrated by patients with xeroderma pigmentosum (XP), a disease caused by inherited defects in NER. XP patients have a 10,000-fold increased risk of skin cancer and early onset neurodegeneration. XP is heterogeneous, ranging from mild to profoundly debilitating. XP severity is proportional to the extent to which NER is disrupted. This suggests that subtle defects in NER, due to, for example, polymorphisms in NER genes, might modestly but significantly impact one’s risk of skin cancer. Since skin cancer affects 20% of Americans and is preventable (by avoiding environmental exposure to UV), identifying those at risk could have a tremendous impact on the health of Americans and healthcare costs. The greatest barrier to identifying those at risk is the lack of an assay to measure NER that is rapid, inexpensive and applicable to samples safely and easily collected from patients. NER occurs in a series of steps involving the recognition of a site of DNA damage, unwinding the DNA locally, excision of a single-stranded oligonucleotide containing the lesion, and templated DNA synthesis to fill the residual gap. NER is the only way that UV-induced photolesions are removed from the genome in human cells. Therefore, NER is measured by the detection and quantification of UV-induced DNA synthesis outside of the S-phase of the cell cycle, or unscheduled DNA synthesis (UDS). Historically, UDS measurement required the use of radioactively-labeled nucleosides and/or specialized equipment. We developed a method to measure NER that employs the thymidine analog 5-ethynyl-2'-deoxyuridine and Click-iT chemistry for fluorescent detection of UDS by flow cytometry. This can be applied to peripheral blood cells for rapid measurement of NER requiring minimally invasive sample collection. UDS in XP patients ranges from <10% to 50%. Nothing is known about the health implications of having a UDS between 50-100%, or how to define 100% NER capacity. This project aims to correct these gaps in knowledge through optimization of our functional assay and proof-of- concept pilot human studies. The assay will be applied to existing cohorts of patients seen at the University of Miami Skin Cancer Clinics, the NIH Undiagnosed Diseases Program or enrolled in the University of Maryland Amish Longevity Study, to interrogate associations between NER capacity and high risk of skin cancer, early onset neurodegeneration, and within family pedigrees, respectively. This project will yield an NER assay applicable to larger population studies aimed at testing associations between NER capacity, environmental exposures and disease risk, and begin to define “normal” NER capacity. The assay could have a significant impact on how risk of squamous cell or basal cell carcinoma of the skin, melanoma, lung or head and neck cancer, neurodegeneration, and resistance to cancer chemotherapy is identified and managed.

项目概要核苷酸切除修复 (NER) 是一种 DNA 修复机制，可识别并去除庞大的螺旋扭曲片段 NER 的关键底物是紫外线 (UV) 辐射引起的损伤。环境暴露于阳光下会产生内源性 DNA 氧化损伤。着色性干皮病 (XP) 患者就是一个戏剧性的例子，这是一种由遗传性缺陷引起的疾病 NER.XP 患者患皮肤癌和早发性神经退行性疾病的风险增加 10,000 倍。 XP 的严重程度与程度成正比，范围从轻微到严重。 NER 被破坏，这表明 NER 中存在细微的缺陷，例如由于 NER 基因的多态性，可能会适度但显着地影响一个人患皮肤癌的风险，因为 20% 的美国人患有皮肤癌。并且是可以预防的（通过避免环境暴露于紫外线），识别处于危险中的人可能会产生巨大的影响对美国人健康和医疗费用的影响是识别高危人群的最大障碍。缺乏快速、廉价且适用于安全、轻松收集样本的 NER 测定方法 NER 发生在一系列涉及 DNA 损伤位点识别、解开 DNA 损伤的步骤中。局部 DNA，切除含有病变的单链寡核苷酸，以及模板化 DNA 合成填补剩余的空白是从基因组中去除紫外线引起的光损伤的唯一方法。因此，NER 是通过检测和定量 UV 诱导的 DNA 合成来测量的。细胞周期 S 期之外，或非计划 DNA 合成 (UDS) 历史上，UDS 测量。需要使用放射性标记的核苷和/或专用设备我们开发了一种方法。使用胸苷类似物 5-乙炔基-2'-脱氧尿苷和 Click-iT 化学进行荧光测量 NER 通过流式细胞术检测 UDS 这可应用于外周血细胞的 NER 快速测量。 XP 患者中需要微创样本采集的情况范围为 <10% 至 50%。关于 UDS 介于 50-100% 之间对健康的影响，或如何定义 100% NER 容量。该项目旨在通过优化我们的功能分析和证明来纠正这些知识差距该试验将应用于伦敦大学现有的患者群体。迈阿密皮肤癌诊所、NIH 未确诊疾病计划或就读于马里兰大学阿米什长寿研究，旨在探讨 NER 能力与早期皮肤癌高风险之间的关系该项目将分别进行 NER 检测。适用于更大的人口研究，旨在测试 NER 能力、环境之间的关联暴露和疾病风险，并开始定义“正常”NER 能力。该测定可能具有显着的效果。对皮肤鳞状细胞癌或基底细胞癌、黑色素瘤、肺癌或头颈癌风险的影响癌症、神经退行性变和癌症化疗耐药性得到识别和管理。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cross-sectional study of factors influencing specialty choice among diverse medical students pursuing careers in dermatology.

对影响从事皮肤科职业的不同医学生专业选择的因素进行横断面研究。

DOI：
10.1016/j.jaad.2023.05.091
发表时间：
2023
期刊：
Journal of the American Academy of Dermatology
影响因子：
13.8
作者：
Smith,Ambrosia;Fulk,Travis;Rypka,Katelyn;Gaddis,Kevin;Farah,Ronda;Mansh,Matthew
通讯作者：
Mansh,Matthew

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