Nevus associated microRNAs as mediators of BRAF-induced growth arrest and biomarkers of melanoma progression

痣相关的 microRNA 作为 BRAF 诱导的生长停滞的介质和黑色素瘤进展的生物标志物

• 批准号: 10474476
• 负责人: Robert Laird Judson-Torres
• 金额: $ 34.19万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-24 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10474476
• 关键词: Address; Anatomy; Area; Automobile Driving; BRAF gene; Benign; Biological Assay; Biological Models; Biology; Biopsy; CDKN2A gene; Cell Cycle; Cells; Chemoprevention; Classification; Clinic; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Cutaneous Melanoma; Data; Data Set; Dermatologist; Detection; Development; Diagnosis; Disease; Drug Delivery Systems; Early Diagnosis; Engineering; Environment; Event; G2/M Arrest; Genes; Genetic; Genetically Engineered Mouse; Genomics; Goals; Growth; Histologic; Human; Human Engineering; In Vitro; Incidence; Individual; Interruption; Knowledge; Lesion; MAP Kinase Gene; Malignant - descriptor; Measurement; Mediating; Mediator of activation protein; Melanocytic nevus; Methods; MicroRNAs; Modeling; Mole the mammal; Molecular; Molecular Profiling; Mutation; Neoplasms; Nevi and Melanomas; Nevus; Non-Invasive Lesion; Oncogenes; PTEN gene; Pathologist; Patients; Physicians; Pigments; Pilot Projects; Positioning Attribute; Prevention; Regulation; Reporting; Reproducibility; Role; Sampling; Signal Transduction; Skin Cancer; Specificity; Survival Rate; System; Techniques; Testing; Training; Transcript; Tumor Suppressor Proteins; Validation; aurora B kinase; base; benign state; cohort; diagnostic accuracy; differential expression; driver mutation; early screening; high risk; improved; in vivo; in vivo Model; in vivo evaluation; inhibitor; innovation; mRNA Expression; melanocyte; melanoma; melanoma biomarkers; melanomagenesis; miRNA expression profiling; novel; novel strategies; pre-clinical; prevent; screening; senescence; skin lesion; transcriptomics; tumor

Summary A single driver mutation, BRAFV600E, drives half of all melanomas. However, in the majority of cases, acquisition of BRAFV600E instead drives benign tumors, such as melanocytic nevi (the common mole). We seek to decipher the intracellular mechanisms that prevent full transformation, and harness this knowledge to develop candidate early diagnosis and chemoprevention strategies. We have discovered a signature of microRNAs (miRNA) as the most differentially expressed transcripts distinguishing nevi from either normal melanocytes or melanomas. We have reported that expression of these miRNAs classifies biopsied pigmented neoplasms with high diagnostic accuracy. To determine whether knowledge of these miRNAs could aid in the prevention of melanoma: First, we conducted a comprehensive identification and functional screen of the targets of the most predictive miRNA, MIR211-5p. Using freshly isolated and CRISPR engineered human normal, nevus and melanoma melanocytes, we identified inhibition of AURKB expression as a critical mechanism driving both BRAFV600E and MIR211-5p associated growth arrest in vitro. Therefore, in Aim 1, our objective is to assess the roll of the MIR211-5p/AURKB axis in nevus formation and transformation in vivo, and the efficacy of disrupting this axis in melanoma chemoprevention. Second, we generated a non-invasive assay for miRNA screening of pigmented lesions prior to biopsy. In a small pilot study, we found the high accuracy of classification of melanocytic neoplasia was retained. In Aim 2, our objective is to validate the utility of using non-invasive profiling of the miRNA signature to screen pigmented skin lesions. Three advances distinguish our proposal. The first is the reproducibility of our miRNA classifier, currently validated on six independent datasets. Second, our model systems for this disease—engineered primary human melanocytes, nevi, and melanomas, combined with an in vivo system that recapitulates both the genetics and progression of melanoma—puts us in a unique position to control for context-specific effects when studying these critical events. Third, is our development of a non-invasive miRNA profiling assay, a molecular profiling technique that is both non-invasive and lesion-specific. Our team, consisting of experts in both the basic biology of miRNA and melanoma, in vivo models of melanoma, topical drug delivery, and the daily practice of melanoma surveillance, allows us to comprehensively tackle this project. This project has both basic and clinical potential significance. Our studies explore novel explanations for nevus initiation, driven by observations made from clinical lesions. We expect these studies to directly result in an increase in the early detection of melanomas and preclinical validation of a strategy for topical chemoprevention for particularly high-risk individuals and/oror anatomic areas.

概括 单一驱动突变 BRAFV600E 驱动了一半的黑色素瘤。然而，在大多数情况下， 相反，获得 BRAFV600E 会导致良性肿瘤，例如黑素细胞痣（常见的痣）。我们寻求 破译阻止完全转化的细胞内机制，并利用这些知识 制定候选早期诊断和化学预防策略。我们发现了一个签名 microRNA (miRNA) 作为区分痣与正常痣的差异最大的转录本 黑色素细胞或黑色素瘤。我们已经报道，这些 miRNA 的表达对活检的色素进行分类 肿瘤诊断准确率高。确定这些 miRNA 的知识是否有助于 预防黑色素瘤：首先，我们对黑色素瘤进行了全面的鉴定和功能筛查。 最具预测性的 miRNA MIR211-5p 的靶点。使用新鲜分离和 CRISPR 工程改造的人类 正常、痣和黑色素瘤黑色素细胞中，我们确定抑制 AURKB 表达是一个关键因素 体外驱动 BRAFV600E 和 MIR211-5p 相关生长停滞的机制。因此，在目标 1 中，我们的 目的是评估 MIR211-5p/AURKB 轴在体内痣形成和转化中的滚动，以及 破坏该轴在黑色素瘤化学预防中的功效。其次，我们进行了非侵入性检测 用于活检前色素病变的 miRNA 筛查。在一项小型试点研究中，我们发现了高精度 保留了黑素细胞瘤的分类。在目标 2 中，我们的目标是验证使用 对 miRNA 特征进行非侵入性分析，以筛选色素性皮肤病变。 我们的建议有三个进步。首先是我们的 miRNA 分类器的可重复性，目前 在六个独立数据集上进行了验证。其次，我们针对这种疾病的模型系统——工程化初级 人类黑色素细胞、痣和黑色素瘤，结合体内系统，概括了 黑色素瘤的遗传学和进展——使我们处于控制特定环境效应的独特位置 在研究这些重大事件时。第三，我们开发了一种非侵入性 miRNA 分析检测方法， 非侵入性且针对病变的分子分析技术。我们的团队由以下领域的专家组成 miRNA 和黑色素瘤的基础生物学、黑色素瘤的体内模型、局部药物递送以及 黑色素瘤监测的日常实践使我们能够全面解决这个项目。 该项目具有基础和临床潜在意义。我们的研究探索了对痣的新解释 启动，由临床病变的观察驱动。我们期望这些研究能够直接产生 增加黑色素瘤的早期检测和局部治疗策略的临床前验证 针对特别高危个体和/或解剖区域的化学预防。