Regulation and Role of miR-183-96-182 in Melanocyte Differentiation and Melanoma

miR-183-96-182 在黑色素细胞分化和黑色素瘤中的调节和作用

• 批准号: 8434136
• 负责人: Eva Hernando
• 金额: $ 32.98万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-27 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8434136
• 关键词: 7q31; Aggressive behavior; Behavior; Binding; Bioinformatics; Biological Assay; Cancer Etiology; Cell Line; Cells; ChIP-seq; Data; Development; Developmental Gene; Disease; Epigenetic Process; Evolution; Genomics; Histone Acetylation; Histones; Homeostasis; In Vitro; Incidence; Injection of therapeutic agent; Laboratories; Link; Malignant - descriptor; Malignant Neoplasms; Melanoma Cell; Metastatic Melanoma; MicroRNAs; Molecular; Mus; Nature; Neoplasm Metastasis; Neural Crest; Operative Surgical Procedures; Phenotype; Physiological; Platelet Factor 4; Population; Primary Neoplasm; Process; Promoter Regions; Property; Regulation; Repression; Role; Skin Cancer; Staging; Stem cells; Survival Rate; Tail; Testing; Therapeutic Intervention; Time; Tissue Differentiation; Tissues; Transcriptional Regulation; Transferase; Uncertainty; Veins; Work; base; calin; cancer cell; cancer stem cell; cancer type; embryonic stem cell; genome-wide; human embryonic stem cell; in vivo; interest; melanocyte; melanoma; migration; mortality; mouse model; overexpression; pluripotency; programs; self-renewal; stem; stem cell differentiation; transcription factor; transcriptome sequencing; tumor

DESCRIPTION (provided by applicant): The incidence and mortality of melanoma, the most aggressive form of skin cancer, are rapidly increasing worldwide. The abysmal survival rates stem from the highly metastatic and chemoresistant behavior of these tumors, but the molecular basis of this aggressive phenotype remains unclear. We and others have shown that melanoma cells display stem-cell-like properties: they often express developmental genes, have multi-differentiation potential, and seem to evoke the migratory nature of neural crest stem cells from which melanocytes arise. These observations suggest that alterations in developmental programs within the melanocytic lineage might underlie the malignant evolution of these cells into metastatic melanoma. In fact, metastasis can be conceived as a normal migration program gone awry, and our laboratory has found that a particular cluster of miRNAs (which in general regulate development, differentiation, and tissue homeostasis) is frequently overexpressed in melanoma cell lines and tissues. We have shown that this miR-183-96-182 cluster promotes migration in vitro and metastasis in vivo in part by controlling the expression of MITF, a master regulator of melanocyte differentiation. Intriguingly, we have also found that miR-183-96-182 is highly expressed in embryonic stem cells (ES), and is silenced during in vitro melanocyte differentiation, inversely correlating with MITF levels. Furthermore, our preliminary studies indicate that histone acetylation and Krupple-like factor-4 (KLF4), a canonical pluripotency transcription factor, govern miR-183-96-182 expression in both hESCs and melanoma cells. We hypothesize that miR-183-96-182 repression is required for melanocyte differentiation, and that alterations in this cluster promote melanoma metastasis, perhaps by conferring stem cell properties to melanoma cells. To test these hypotheses, we will first determine the transcriptional and epigenetic regulation of the miR-183-96-182 cluster in physiological and pathological contexts, from human embryonic stem cell stage through melanocyte differentiation and in melanoma cells (Aim 1). In Aim 2 we will test whether miR-183-96-182 and/or KLF4 confer stem-cell-like properties (e.g., self-renewal, multi-differentiation capacity) on melanoma cells through several cell-based assays. In Aim 3, we will determine how modulation of this miRNA cluster and/or KLF4 influence the propensity of melanoma cells for metastasis in mice. Understanding the mechanisms that modulate miR-183-96-182 overexpression, which is clearly involved in metastasis, could provide a foothold for altering the aggressiveness of this very aggressive cancer, and perhaps provide a framework for similar studies in other recalcitrant cancer types.

描述（由申请人提供）：皮肤癌最具侵略性的黑色素瘤的发病率和死亡率正在全球范围内迅速增加。糟糕的存活率源于这些肿瘤的高度转移性和化学抗性行为，但是这种攻击性表型的分子基础尚不清楚。我们和其他人已经表明，黑色素瘤细胞表现出类似干细胞的特性：它们通常表达发育基因，具有多差异潜力，并且似乎唤起了神经rest干细胞的迁移性，而黑素细胞从中产生了黑素细胞。这些观察结果表明，黑色素谱系中发育计划的改变可能是这些细胞的恶性进化为转移性黑色素瘤的基础。实际上，转移可以被认为是正常的迁移程序出了问题，我们的实验室发现，特定的miRNA簇（通常调节发育，分化和组织稳态）在黑色素瘤细胞系和组织中经常过表达。我们已经表明，这种miR-183-96-182簇在体外促进迁移，部分通过控制MITF的表达，MITF的表达是黑素细胞分化的主要调节剂。有趣的是，我们还发现miR-183-96-182在胚胎干细胞（ES）中高度表达，并且在体外黑素细胞分化过程中被沉默，与MITF水平成反比。此外，我们的初步研究表明，组蛋白乙酰化和类似Krupple的因子-4（KLF4）是一种规范的多能转录因子，负责MiR-183-96-182在hESC和黑色素瘤细胞中的表达。我们假设MiR-183-96-182抑制黑色素细胞分化需要抑制作用，并且该簇中的改变促进了黑色素瘤转移，也许是通过将干细胞特性赋予黑色素瘤细胞。为了检验这些假设，我们将首先确定MiR-183-96-182簇在生理和病理环境中的转录和表观遗传调节，从人类胚胎干细胞阶段通过黑素细胞分化和黑色素瘤细胞中的人类胚胎干细胞阶段（AIM 1）。在AIM 2中，我们将通过几种基于细胞的测定方法测试MiR-183-96-182和/或KLF4赋予干细胞样性能（例如自我更新，多差异能力）。在AIM 3中，我们将确定该miRNA簇和/或KLF4的调节如何影响小鼠转移的黑色素瘤细胞倾向。了解调节MiR-183-96-182过表达的机制显然与转移有关，可以为改变这种非常侵略性癌症的侵略性提供立足点，并可能为其他顽固性癌症类型的类似研究提供了框架。