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: 10292583
• 负责人: Robert Laird Judson-Torres
• 金额: $ 34.88万
• 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/10292583
• 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/antagonist; 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 一个驱动器突变，即BRAFV600E，驱动了所有黑色素瘤的一半。但是，在大多数情况下， 相反，BRAFV600E的获取驱动了良性肿瘤，例如黑素细胞Nevi（常见的痣）。我们寻找 解释了防止完全转化的细胞内机制，并利用这些知识 制定候选人的早期诊断和化学预防策略。我们发现了一个签名 microRNA（miRNA）是最不同的转录本，将Nevi与任何一个正常 黑素细胞或黑色素瘤。我们报道了这些miRNA类别活检的表达 具有高诊断精度的肿瘤。确定这些miRNA是否可以帮助 预防黑色素瘤：首先，我们进行了全面的识别和功能屏幕 最预测性miRNA的目标miR211-5p。使用新鲜隔离和CRISPR工程的人 正常，颈和黑色素瘤黑色素细胞，我们确定对Aurkb表达的抑制作用是关键 驱动BRAFV600E和MIR211-5P的机制在体外相关生长停滞。因此，在AIM 1中，我们 目的是评估Mir211-5p/Aurkb轴的滚动，体内和转化中的卷。 在黑色素瘤化学预防中破坏该轴的效率。其次，我们产生了一个非侵入性测定 用于在活检之前对猪病变进行miRNA筛查。在一项小型试点研究中，我们发现 保留了黑色素瘤的分类。在AIM 2中，我们的目标是验证使用的实用性 miRNA签名的非侵入性分析以筛选有色皮肤病变。 三个进步区分了我们的建议。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，我们的该疾病模型系统 - 设计的主要系统 人类黑色素细胞，Nevi和黑色素瘤结合了一个体内系统，可概括这两种系统 黑色素瘤的遗传学和进展 - 使我们处于控制特定效果的独特位置 在研究这些关键事件时。第三，是我们开发非侵入性miRNA分析测定法， 分子分析技术既是非侵入性和病变特异性的。我们的团队，由专家组成 miRNA和黑色素瘤的基本生物学，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练习使我们能够全面解决该项目。 该项目具有基本和临床潜在的意义。我们的研究探讨了奈夫的新颖解释 启动，由临床病变的观察结果驱动。我们希望这些研究将直接导致 早期发现黑色素瘤和临床前验证局部策略 针对特别高风险个体和/或解剖区域的化学预防。