MicroRNA and protein profiling in melanocytes exposed to solar UVR in situ

原位暴露于太阳 UVR 的黑素细胞中的 MicroRNA 和蛋白质分析

• 批准号: 8443934
• 负责人: MLNikki L Harter
• 金额: $ 24.15万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-12-01 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8443934
• 关键词: Affect; Area; Bioinformatics; Biological Markers; Cancerous; Cells; Computer software; Cutaneous Melanoma; Data; Databases; Development; Dose; Drug Targeting; Environmental Carcinogens; Epidemiology; Exposure to; Gene Targeting; Human; Human Volunteers; In Situ; Individual; Learning; Lesion; Measures; Messenger RNA; MicroRNAs; Molecular; Pathway interactions; Patients; Pattern; Persons; Phase; Play; Portraits; Positioning Attribute; Protein Microchips; Proteins; Proteomics; Recording of previous events; Relative (related person); Risk; Risk Factors; Role; Shoulder; Simulate; Skin Cancer; Sunlight; Techniques; Technology; Testing; Tissues; Ultraviolet Rays; innovation; laser capture microdissection; melanocyte; melanoma; novel; novel therapeutics; protein profiling; public health relevance; research study; treatment strategy; volunteer

DESCRIPTION (provided by applicant): For years, epidemiological and scientific studies have supported a role for solar ultraviolet radiation (UVR) in the development of melanoma. Our new data suggests that UV rays can differentially affect the expression of microRNAs (miRNAs) in the melanocytes of healthy persons versus those who have a history of melanoma. We therefore hypothesize that melanoma prone melanocytes, when exposed to solar UVR, differ markedly from healthy melanocytes in the deregulation of miRNAs and that this aberrancy leads to the perturbation of molecular pathways that contribute in part to the development of melanoma. With the use of novel techniques and proteomic technology, we will test this hypothesis by performing the following studies: Aim 1 - Evaluate the expression of miRNAs between the melanocytes of healthy persons and melanoma patients after exposure to solar UVR in situ. Specifically we will: a) Use biologically relevant doses of simulated solar UVR (ssUVR) to irradiate a statistical number of human volunteers (both healthy and melanoma patients), and thereafter isolate irradiated or un-irradiated melanocytes by laser capture microdissection (LCM). b) Quantify relative changes in the expression of miRNAs between the irradiated and un-irradiated melanocytes from each volunteer and then use bioinformatic approaches to identify miRNAs that are commonly deregulated amongst each of the two groups. c) Confirm the expression patterns observed on the microRNA array cards and use currently available software databases to tentatively predict miRNA target genes. d) Perform parallel experiments to profile mRNAs between the same set of irradiated and un-irradiated melanocytes as a first step in identifying candidate UV-responsive miRNA-mRNA relationships. Aim 2 - Identify proteomic pathways potentially affected by aberrantly expressed UV-responsive miRNAs in melanocytes of melanoma patients. Specifically, we will: a) Use reverse phase protein microarrays (RPMA) to measure the quantity of individual proteins (miRNA targets) between the irradiated and un-irradiated melanocytes of melanoma patients or healthy individuals. b) Use this new class of proteomic profiling technology to analyze the potentially altered state of molecular pathways relevant to the early development of melanoma.

描述（由申请人提供）：多年来，流行病学和科学研究都支持太阳紫外线辐射 (UVR) 在黑色素瘤发展中的作用。我们的新数据表明，紫外线对健康人和有黑色素瘤病史的人黑色素细胞中 microRNA (miRNA) 的表达有不同的影响。因此，我们假设，当暴露于太阳 UVR 时，易患黑色素瘤的黑色素细胞在 miRNA 失调方面与健康黑色素细胞显着不同，并且这种异常导致分子途径的扰动，从而在一定程度上促进了黑色素瘤的发展。通过使用新技术和蛋白质组学技术，我们将通过进行以下研究来检验这一假设： 目标 1 - 评估原位暴露于太阳 UVR 后健康人和黑色素瘤患者黑色素细胞之间 miRNA 的表达。具体来说，我们将：a）使用生物学相关剂量的模拟太阳紫外线（ssUVR）来照射统计数量的人类志愿者（健康和黑色素瘤患者），然后通过激光捕获显微切割（LCM）分离受照射或未受照射的黑色素细胞。 b) 量化每位志愿者经辐射和未经辐射的黑素细胞之间 miRNA 表达的相对变化，然后使用生物信息学方法来识别两组中通常失调的 miRNA。 c) 确认在 microRNA 阵列卡上观察到的表达模式，并使用当前可用的软件数据库初步预测 miRNA 靶基因。 d) 进行平行实验以分析同一组受辐射和未受辐射黑素细胞之间的 mRNA，作为识别候选 UV 响应 miRNA-mRNA 关系的第一步。目标 2 - 确定黑色素瘤患者黑色素细胞中异常表达的紫外线响应 miRNA 可能影响的蛋白质组通路。具体来说，我们将： a) 使用反相蛋白微阵列 (RPMA) 测量黑色素瘤患者或健康个体经辐射和未经辐射的黑色素细胞之间的单个蛋白质（miRNA 靶标）的数量。 b) 使用这种新型蛋白质组分析技术来分析与黑色素瘤早期发展相关的分子途径的潜在改变状态。