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增加表明， 除了目前的紫外线防护方法外，还需要其他方法。我们的长期目标是发展这种预防 基于对正常人性别差异DNA损伤修复的全面了解的策略 人类黑素细胞本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黑素细胞的DNA损伤修复。我们初步的流行病学数据 唾液睾酮（T）水平和T/E2（雌二醇）水平与黑色素瘤风险呈负相关; 而我们的细胞生物学数据显示，在培养基中添加睾酮可以增强正常人的DNA修复， 人类黑素细胞我们的中心假设是，较高的睾酮水平，或较高的T/E2比值， 来自涉及PARP 1的UV诱导的DNA损伤的NHM。我们还假设，来自男性和 女性需要不同的T/E2平衡，因为T/E2水平在两性中明显不同。两 具体目标1：确定DNA损伤反应中的T和T/E2效应 男性和女性的NHM（体内研究）。将招募人类受试者，并将用以下药物处理皮肤： 在不存在和存在局部应用睾酮的情况下的太阳模拟UV辐射（以改变T或 T/E2比率）。将监测DNA损伤标志物、激素受体（AR和GPER 1）和PARP 1水平 在不同的条件下，将比较标记物阳性细胞的时间进程和百分比， 黑素细胞和其他皮肤细胞。角质形成细胞-黑素细胞的相互作用受性激素的影响， 检测TNFα和TNFR 1。具体目的2：测试T/E2在来源于以下的培养黑素细胞中的作用： 男性或女性起源，并确定性激素调节的DNA修复途径在体外。激素 通过在UVR处理之前以各种比例加入激素来检测效果。基因敲低 或AR特异性抑制剂或激动剂将用于操纵T/AR和E2/GPER 1，和 DNA损伤反应和信号通路将被检查。创新：据我们所知， 这项研究是第一个假设T和E2平衡对于T/AR预防皮肤黑色素瘤至关重要的研究 系统通过增强DNA修复能力作为肿瘤抑制因子。拟议的研究具有重要意义 因为它有望通过增加新的参数， 进行性激素测试以评估风险，或通过饮食或补充剂调节激素水平以预防。如果 成功的，它有望导致一个范式的变化，从目前的预防“避免紫外线辐射”到未来 “避免紫外线辐射加上荷尔蒙平衡”。