A role of balanced sex hormone in DNA repair in human melanocytes

平衡性激素在人类黑素细胞 DNA 修复中的作用

• 批准号: 10666307
• 负责人: Feng Liu-Smith
• 金额: $ 42.35万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-25 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10666307
• 关键词: Adult; Age; Agonist; Androgen Receptor; Antibodies; Apoptosis; Area; Behavior; Biological; Biological Process; Biopsy; Biopsy Specimen; Biphasic Pattern; Blood; Blood specimen; Cell Culture Techniques; Cell Cycle Arrest; Cell Cycle Progression; Cells; Cellular biology; Culture Media; Cutaneous Melanoma; DNA Damage; DNA Repair; DNA Repair Pathway; Data; Dermal; Development; Diet; Dose; Epidemiology; Equilibrium; Estradiol; Etiology; Exposure to; Female; Fibroblasts; Functional disorder; Future; Genes; Goals; Gonadal Steroid Hormones; Hormonal; Hormone Receptor; Hormones; Human; Immunofluorescence Immunologic; In Vitro; Incidence; Individual; Investigation; Langerhans cell; Link; Logistic Regressions; Malignant Neoplasms; Measurement; Menopause; Metabolism; Methods; Molecular; Molecular and Cellular Biology; Monitor; Penetration; Predisposition; Prevention; Prevention strategy; Public Health Education; Radiation Induced DNA Damage; Radiation induced damage; Radiation therapy; Risk; Risk Assessment; Risk Factors; Role; Safety; Saliva; Salivary; Sex Differences; Sex Differentiation; Signal Pathway; Signal Transduction; Skin; Stanolone; Statistical Data Interpretation; System; TNF gene; TNFRSF1A gene; Testing; Testosterone; Time; Topical application; Tumor Suppressor Proteins; UV Radiation Exposure; UV induced; UV induced DNA damage; Ultraviolet Rays; United States; Woman; age related; anticancer research; cancer risk; cell type; epidemiologic data; epidemiology study; hormone regulation; human old age (65+); human subject; in vitro Model; in vivo; inhibitor; innovation; keratinocyte; knock-down; male; melanocyte; melanoma; men; novel; older men; older women; overexpression; recruit; reproductive; response; saliva sample; sex; young man; young woman

The continuous increase in incidence rates of melanoma suggests novel prevention strategies may be needed in addition to current UVR-avoidance methods. Our long-term goal is to develop such prevention strategies based on a comprehensive understanding of the sex differentiated DNA damage repair in normal human melanocytes. The objective of this R21 proposal is to determine the sex hormonal regulation of UVR- induced DNA damage repair in human melanocytes in vitro and in vivo. Our preliminary epidemiology data showed that salivary testosterone (T) level and T/E2 (estradiol) levels is inversely correlated with melanoma risk; while our cell biology data showed that addition of testosterone in culture media enhanced DNA repair in normal human melanocytes. Our central hypothesis is that higher testosterone levels, or higher T/E2 ratios, protect NHMs from UV-induced DNA damage involving PARP1. We also hypothesize that NHMs from males and females require a different balanced T/E2, as the T/E2 levels apparently dramatically differ in the two sexes. Two specific aims are proposed: Specific Aim 1: To determine the T and T/E2 effects in DNA damage responses in NHMs in men and women (in vivo study). Human subjects will be recruited, and skin will be treated with solar simulated UV radiation in the absence and presence of topical application of testosterone (to alter T or T/E2 ratios). DNA damage markers, hormone receptors (AR and GPER1) and PARP1 levels will be monitored following a time course and percentage of marker positive cells will be compared in different conditions in melanocytes and other skin cells. The keratinocytes-melanocytes interaction impacted by sex hormones will be examined by TNFα and TNFR1. Specific Aim 2: To test T/E2 effect in cultured melanocytes derived from male or female origins and to determine sex hormone-modulated DNA repair pathway in vitro. Hormonal effect will be detected by inclusion of the hormones at various ratios before UVR treatment. Gene knockdown of AR and GPER1, or AR-specific inhibitors or agonists will be used to manipulate T/AR and E2/GPER1, and DNA damage responses and signal pathways will be examined. The innovation: to the best of our knowledge, this study is the first to hypothesize a balanced T and E2 is critical for skin melanoma prevention with T/AR system acting as a tumor suppressor through enhancing DNA repair capacity. The proposed study is significant because it is expected to make a paradigm change in melanoma prevention by adding new parameters such as sex hormone test for risk assessment, or adjusting hormone levels through diet or supplement for prevention. If successful, it is expected to lead to a paradigm change of the prevention from current “avoiding UVR” to future “avoiding UVR plus hormone balance”.

黑色素瘤发病率的持续提高表明，新型预防策略可能是 除了当前的UVR避免方法。我们的长期目标是开发这种预防 基于对性别分化的DNA损伤修复的全面理解的策略 人类黑色素细胞。该R21提案的目的是确定UVR-的性激素调节 在体外和体内诱导人类黑素细胞的DNA损伤修复。我们的初步流行病学数据 表明唾液睾丸激素（T）水平和T/E2（雌二醇）水平与黑色素瘤风险成反比。 虽然我们的细胞生物学数据表明，在培养基中添加睾丸激素在正常情况下增强了DNA修复 人类黑色素细胞。我们的中心假设是较高的睾丸激素水平或更高的T/E2比率保护 来自紫外线诱导的DNA损伤的NHM涉及PARP1。我们还假设来自男性和 女性需要不同的平衡T/E2，因为T/E2水平在两个性别中显然有很大的不同。二 提出了具体目的：具体目的1：确定DNA损伤反应中的T和T/E2效应 在男性和女人的NHM中（体内研究）。人类受试者将被招募，皮肤将被治疗 在不存在和存在局部应用睾丸激素的情况下，太阳能模拟紫外线辐射（改变T或 T/E2比率）。将监测DNA损伤标记物，马酮受体（AR和GPER1）和PARP1水平 在时间过程和标记阳性细胞的百分比之后，将在不同条件下进行比较 黑色素细胞和其他皮肤细胞。性激素影响的角质形成细胞 - 核细胞相互作用将是 由TNFα和TNFR1检查。特定目的2：测试T/E2效应在培养的黑素细胞中 男性或女性起源，并在体外确定性别同性调节的DNA修复途径。激素 在UVR治疗之前，将通过在各种ratos中纳入马匹来检测效果。基因敲低 AR和GPER1或AR特异性抑制剂或激动剂将用于操纵T/AR和E2/GPER1，并且 将检查DNA损伤响应和信号途径。创新：据我们所知， 这项研究是第一个假设平衡T和E2的研究对于预防皮肤黑色素瘤至关重要 通过增强DNA修复能力充当肿瘤抑制器的系统。拟议的研究很重要 因为预计通过添加新参数（例如 性荷花测试进行风险评估，或通过饮食或补充剂进行预防调整霍斯奈人的水平。如果 成功，预计将导致从当前的“避免UVR”到未来的预防变化范式 “避免使用UVR加上同性恋平衡”。