整个黑色素瘤基因组的高突变率对从背景突变中区分真正的驱动事件提出了重大挑战。许多重复出现的非编码改变，例如增强子中的改变，可以塑造肿瘤的进化，从而强调了系统破译黑色素瘤中增强子破坏的重要性。在这里，我们利用 297 个黑色素瘤全基因组测序样本来优先考虑高度重复的区域。通过对黑色素瘤细胞中高度重复的区域相关增强子进行基因组规模的 CRISPR 干扰 (CRISPRi) 筛选，我们鉴定了 66 个可能具有肿瘤抑制作用的重要命中。这些功能增强子显示出独特的突变模式，独立于黑色素瘤中经典的显着突变基因。对必需增强子的靶基因分析揭示了黑色素瘤生长的许多已知和隐藏的机制。利用广泛的功能验证实验，我们证明超级增强子元件可以通过靶向 MEF2A 来调节黑色素瘤细胞增殖，而另一个远端增强子能够通过长程相互作用维持 PTEN 肿瘤抑制潜力。我们的研究建立了关键增强子和其在黑色素瘤生长和进展中的靶基因，并阐明了黑色素瘤驱动基因失调的新机制和新的治疗靶向策略的识别。© 2023。作者。

The high mutation rate throughout the entire melanoma genome presents a major challenge in stratifying true driver events from the background mutations. Numerous recurrent non-coding alterations, such as those in enhancers, can shape tumor evolution, thereby emphasizing the importance in systematically deciphering enhancer disruptions in melanoma.Here, we leveraged 297 melanoma whole-genome sequencing samples to prioritize highly recurrent regions. By performing a genome-scale CRISPR interference (CRISPRi) screen on highly recurrent region-associated enhancers in melanoma cells, we identified 66 significant hits which could have tumor-suppressive roles. These functional enhancers show unique mutational patterns independent of classical significantly mutated genes in melanoma. Target gene analysis for the essential enhancers reveal many known and hidden mechanisms underlying melanoma growth. Utilizing extensive functional validation experiments, we demonstrate that a super enhancer element could modulate melanoma cell proliferation by targeting MEF2A, and another distal enhancer is able to sustain PTEN tumor-suppressive potential via long-range interactions.Our study establishes a catalogue of crucial enhancers and their target genes in melanoma growth and progression, and illuminates the identification of novel mechanisms of dysregulation for melanoma driver genes and new therapeutic targeting strategies.© 2023. The Author(s).