Mapping DNA Repair and Error-Prone DNA Synthesis in Geriatric Skin

绘制老年皮肤 DNA 修复和容易出错的 DNA 合成图谱

• 批准号: 10253181
• 负责人: Michael George Kemp
• 金额: --
• 依托单位: DAYTON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10253181
• 关键词: ATR gene; Address; Affect; Age; Age-Years; Aging; Apical; Biology; Biopsy; Caring; Cause of Death; Cell Cycle Progression; Cells; Clinic; Clinical; Colon; Colon Carcinoma; DNA; DNA Damage; DNA Repair; DNA biosynthesis; DNA mapping; DNA photoproducts; DNA-Directed DNA Polymerase; Data; Defect; Dermal; Dermatology; Development; Diagnosis; Elderly; Epidermis; Epithelial; Erythrocytes; Excision; Excision Repair; Exposure to; Failure; Fibroblasts; Financial Hardship; General Population; Genome; Genomic DNA; Genomics; Goals; Growth Factor; Healthcare Systems; Hormones; Human; In Vitro; Incidence; Individual; Injections; Insulin; Insulin-Like Growth Factor I; Insulin-Like-Growth Factor I Receptor; Intervention; Knowledge; Lasers; Lead; Lung; Maintenance; Malignant Neoplasms; Malignant neoplasm of esophagus; Malignant neoplasm of lung; Modeling; Morbidity - disease rate; Mutagenesis; Neoplasms; Nucleotides; Organ Transplantation; Outcome; Pathogenesis; Pathway interactions; Phenotype; Phosphotransferases; Physiological; Play; Population; Production; Property; Publishing; Receptor Activation; Receptor Signaling; Rejuvenation; Research; Resolution; Risk Factors; Role; Signal Transduction; Skin; Skin Cancer; Skin Carcinogenesis; Skin Carcinoma; Solid; Source; Stress Response Signaling; Structure; Sun Exposure; Sunlight; Testing; Tissues; Transplant Recipients; Treatment Cost; UV Mutagenesis; UV Radiation Exposure; UV carcinogenesis; UV induced; UVB induced; Ultraviolet B Radiation; Ultraviolet Rays; Veterans; Work; age related; aged; base; cancer initiation; cancer risk; carcinogenesis; cost; cytokine; dietary; dimer; experience; genome-wide analysis; genomic locus; human disease; improved; in vivo; innovation; keratinocyte; military veteran; mortality; novel; novel therapeutic intervention; patient population; photolesion; premalignant; prevent; programs; repaired; replication stress; response; senescence; skin disorder; skin wound; tumorigenesis; tumorigenic; ultraviolet; young adult; ATR gene; Address; Affect; Age; Age-Years; Aging; Apical; Biology; Biopsy; Caring; Cause of Death; Cell Cycle Progression; Cells; Clinic; Clinical; Colon; Colon Carcinoma; DNA; DNA Damage; DNA Repair; DNA biosynthesis; DNA mapping; DNA photoproducts; DNA-Directed DNA Polymerase; Data; Defect; Dermal; Dermatology; Development; Diagnosis; Elderly; Epidermis; Epithelial; Erythrocytes; Excision; Excision Repair; Exposure to; Failure; Fibroblasts; Financial Hardship; General Population; Genome; Genomic DNA; Genomics; Goals; Growth Factor; Healthcare Systems; Hormones; Human; In Vitro; Incidence; Individual; Injections; Insulin; Insulin-Like Growth Factor I; Insulin-Like-Growth Factor I Receptor; Intervention; Knowledge; Lasers; Lead; Lung; Maintenance; Malignant Neoplasms; Malignant neoplasm of esophagus; Malignant neoplasm of lung; Modeling; Morbidity - disease rate; Mutagenesis; Neoplasms; Nucleotides; Organ Transplantation; Outcome; Pathogenesis; Pathway interactions; Phenotype; Phosphotransferases; Physiological; Play; Population; Production; Property; Publishing; Receptor Activation; Receptor Signaling; Rejuvenation; Research; Resolution; Risk Factors; Role; Signal Transduction; Skin; Skin Cancer; Skin Carcinogenesis; Skin Carcinoma; Solid; Source; Stress Response Signaling; Structure; Sun Exposure; Sunlight; Testing; Tissues; Transplant Recipients; Treatment Cost; UV Mutagenesis; UV Radiation Exposure; UV carcinogenesis; UV induced; UVB induced; Ultraviolet B Radiation; Ultraviolet Rays; Veterans; Work; age related; aged; base; cancer initiation; cancer risk; carcinogenesis; cost; cytokine; dietary; dimer; experience; genome-wide analysis; genomic locus; human disease; improved; in vivo; innovation; keratinocyte; military veteran; mortality; novel; novel therapeutic intervention; patient population; photolesion; premalignant; prevent; programs; repaired; replication stress; response; senescence; skin disorder; skin wound; tumorigenesis; tumorigenic; ultraviolet; young adult

PROJECT SUMMARY/ABSTRACT Non-melanoma skin cancers (NMSCs) are a growing problem among our nation’s Veteran population, and current treatments are both costly and potentially disfiguring. The primary risk factors for NMSC development are exposure to ultraviolet B (UVB) wavelengths of sunlight and advanced age. Though the mutagenic effects of UVB-induced bipyrimidine dimers in genomic DNA are well known, the mechanisms by which age impacts UVB- dependent NMSC initiation in skin keratinocytes are less clear. The unique structural and physiological properties of geriatric skin likely influence both the number and distribution of photolesions that form across the genome of basal keratinocytes after UV exposure and the ways in which these cells respond to the damage. Moreover, aged tissues including skin are known to contain a higher number of senescent dermal fibroblasts than younger skin, which may alter cytokine and growth factor production and contribute to a pro-tumorigenic microenvironment. One such factor that is expressed at lower levels in geriatric skin is insulin-like growth factor 1 (IGF-1). Our published and preliminary data show that the loss of IGF-1/IGF-1 receptor (IGF-1R) signaling in epidermal keratinocytes results in UVB-irradiated keratinocytes that are unable to efficiently remove UVB photoproducts from DNA or to properly activate the apical DNA damage response kinase ATR, which functions to suppress error-prone DNA synthesis on damaged DNA templates. The objective of this proposal is to determine how age and specifically IGF-1 receptor activation affect the formation and replication of UVB DNA photoproducts in basal epidermal keratinocytes in human skin. The background of the research team, which includes expertise in DNA repair, replication stress response signaling, genomics, keratinocyte biology, and clinical dermatology, makes this group uniquely qualified to address this important clinical issue. The central hypothesis of this proposal is that the skin of geriatric individuals is prone to carry out a more mutagenic form of DNA synthesis following UVB exposure than the skin of younger individuals. This hypothesis has been derived from significant preliminary and published data with both cultured keratinocytes in vitro and human skin biopsies ex vivo and in vivo. The rationale for this proposed research is that it will define how the unique features of geriatric skin predispose it to UVB mutagenesis and carcinogenesis. Our hypothesis will be tested by carrying out the following three specific aims: Aim 1) Define how age and IGF-1 status influence UVB-induced DNA photoproduct formation and repair across the keratinocyte genome in human skin epidermis; Aim 2) Determine how IGF-1 signaling impacts the utilization of error-prone DNA polymerases in UVB-irradiated human skin explants ex vivo and cultured keratinocytes in vitro; and Aim 3) Define how subject age and IGF-1 status affect the replication of UVB-damaged DNA in human skin epidermis in vivo. Our approach is innovative because it will provide a detailed, quantitative characterization of DNA damage formation and replication in skin epidermis using novel technical approaches that have not previously been applied to human skin. The proposed research is significant to the VA because it will show how the geriatric skin microenvironment contributes to UV mutagenesis and carcinogenesis. Ultimately, we expect that the knowledge that will be gained here will provide further mechanistic rationale for the use of clinical skin rejuvenation interventions aimed at preventing NMSC in specific patient populations. Lastly, because many other cancers are associated with both advanced age and the exposure of epithelial tissues to environmental and dietary DNA damaging agents, our work likely has implications for understanding the pathogenesis of lung, colon, and esophageal cancers in older veterans.

项目摘要/摘要 非黑色素瘤皮肤癌（NMSC）是我们国家资深人口中日益增长的问题， 当前的治疗既昂贵又可能毁容。 NMSC开发的主要风险因素 是暴露于阳光和高龄的紫外线B（UVB）波长。虽然诱变的影响 UVB诱导的基因组DNA中的双吡啶二聚体是众所周知的，年龄会影响UVB-的机制 皮肤角质形成细胞中的依赖性NMSC启动不太清楚。独特的结构和物理特性 老年皮肤可能会影响整个基因组形成的照片的数量和分布 紫外线暴露后的基底角质形成细胞以及这些细胞对损伤做出反应的方式。而且， 已知包括皮肤在内的老年组织包含比年轻人更高的感觉皮肤成纤维细胞 皮肤，可能会改变细胞因子和生长因子的产生，并有助于促肿瘤 微环境。在老年皮肤中较低水平表达的一个因素是胰岛素样生长因子 1（IGF-1）。我们发布的初步数据表明，IGF-1/IGF-1受体（IGF-1R）信号的损失 表皮角质形成细胞导致无法有效去除UVB的UVB辐照角质形成细胞 来自DNA的光产物或正确激活顶端DNA损伤反应激酶ATR，该响应激酶ATR功能 在受损的DNA模板上抑制容易发生的DNA合成。该提议的目的是 确定年龄和特别是IGF-1受体激活如何影响UVB DNA的形成和复制 人皮肤表皮角质形成细胞的光产物。研究团队的背景 包括DNA修复，复制应力反应信号传导，基因组学，角质形成细胞生物学和 临床皮肤病学使该组具有独特的资格来解决这一重要的临床问题。中央 该提议的假设是老年人的皮肤容易执行更诱变的形式 UVB暴露后的DNA合成比年轻个体的皮肤。该假设已得出 从体外和人类皮肤活检中具有培养的角质形成细胞的重要初步和已发表的数据中 前体内和体内。这项拟议研究的理由是，它将定义 老年皮肤使其易于使用UVB诱变和癌变。我们的假设将通过携带来检验 解决以下三个特定目标：目标1）定义年龄和IGF-1状态如何影响UVB诱导的DNA 人类皮肤表皮中角质形成细胞基因组的光产物形成和修复；目标2）确定 IGF-1信号如何影响UVB辐射的人体皮肤中容易发生的DNA聚合酶的利用 外植体在体外体内和培养的角质形成细胞；目标3）定义主题年龄和IGF-1状态如何影响 体内人类皮肤表皮中UVB受损的DNA的复制。我们的方法是创新的，因为它 将提供对皮肤表皮中DNA损伤形成和复制的详细定量表征 使用以前尚未应用于人类皮肤的新型技术方法。拟议的研究 对VA意义重大，因为它将显示老年皮肤微环境如何有助于紫外线 诱变和癌变。最终，我们期望在这里获得的知识将提供 用于使用临床皮肤修订干预措施的进一步的机械原理，以防止NMSC 特定的患者人群。最后，因为许多其他癌症都与高龄和 上皮组织暴露于环境和饮食DNA破坏剂中，我们的工作可能具有 了解老年退伍军人中肺，结肠和食管癌的发病机理的意义。