Department of Molecular Biology, School of Biological Sciences, University of California San Diego, La Jolla, California, United States of America.
PLoS Comput Biol. 2024 Jul 30;20(7):e1012271. doi: 10.1371/journal.pcbi.1012271. eCollection 2024 Jul.
Melanoma showcases a complex interplay of genetic alterations and intra- and inter-cellular morphological changes during metastatic transformation. While pivotal, the role of specific mutations in dictating these changes still needs to be fully elucidated. Telomerase promoter mutations (TERTp mutations) significantly influence melanoma's progression, invasiveness, and resistance to various emerging treatments, including chemical inhibitors, telomerase inhibitors, targeted therapy, and immunotherapies. We aim to understand the morphological and phenotypic implications of the two dominant monoallelic TERTp mutations, C228T and C250T, enriched in melanoma metastasis. We developed isogenic clonal cell lines containing the TERTp mutations and utilized dual-color expression reporters steered by the endogenous Telomerase promoter, giving us allelic resolution. This approach allowed us to monitor morpholomic variations induced by these mutations. TERTp mutation-bearing cells exhibited significant morpholome differences from their wild-type counterparts, with increased allele expression patterns, augmented wound-healing rates, and unique spatiotemporal dynamics. Notably, the C250T mutation exerted more pronounced changes in the morpholome than C228T, suggesting a differential role in metastatic potential. Our findings underscore the distinct influence of TERTp mutations on melanoma's cellular architecture and behavior. The C250T mutation may offer a unique morpholomic and systems-driven advantage for metastasis. These insights provide a foundational understanding of how a non-coding mutation in melanoma metastasis affects the system, manifesting in cellular morpholome.
黑色素瘤在转移转化过程中表现出遗传改变以及细胞内和细胞间形态变化的复杂相互作用。虽然至关重要，但特定突变在决定这些变化中的作用仍需要充分阐明。端粒酶启动子突变（TERTp 突变）显著影响黑色素瘤的进展、侵袭性以及对各种新兴治疗方法的耐药性，包括化学抑制剂、端粒酶抑制剂、靶向治疗和免疫疗法。我们旨在了解在富含黑色素瘤转移的两种主要单等位基因 TERTp 突变 C228T 和 C250T 中的形态和表型影响。我们开发了含有 TERTp 突变的同基因克隆细胞系，并利用由内源性端粒酶启动子驱动的双色表达报告基因，实现了等位基因分辨率。这种方法使我们能够监测这些突变诱导的形态变化。与野生型相比，携带 TERTp 突变的细胞表现出明显的形态差异，表现出增加的等位基因表达模式、增强的伤口愈合率和独特的时空动力学。值得注意的是，C250T 突变比 C228T 突变在形态变化上表现出更为显著的差异，表明其在转移潜能方面存在差异作用。我们的研究结果强调了 TERTp 突变对黑色素瘤细胞结构和行为的独特影响。C250T 突变可能为转移提供了独特的形态和系统驱动优势。这些发现为了解黑色素瘤转移中非编码突变如何影响系统，表现为细胞形态变化提供了基础理解。
