Understanding spatial and temporal clonal evolution of melanoma and its niche under treatment pressure

黑色素瘤对于转移性疾病具有不良预后。尽管其治疗有所改进，但约50%的患者仍会产生耐药性，并伴有广泛的复发和极高的死亡率。越来越多的证据表明，高度的肿瘤内异质性与患者疾病预后不良相关。尽管通过基因组和表型研究来解开克隆动态的大的努力，我们的理解仍然是基本的。因此，该项目采取了一种更可控的方法，研究不同的黑色素瘤克隆在靶向治疗之前和期间如何竞争和不同的脆弱性。最终目的是获得关于如何降低耐药黑色素瘤克隆的适应性的机制见解，从而实现更持久的治疗反应。我将用一种新的方法产生肿瘤内异质性，该方法随机引入定义的临床相关致癌突变，再加上肿瘤内独特的荧光团，可以使用活体成像在体内跟踪。独特的荧光团将使离体分离不同的克隆和下游机制分析的变化，信号和基因表达。此外，我将研究当具有特定突变的克隆扩增时肿瘤微环境如何变化。通过这些研究获得的知识将被用来测试更好的体内治疗策略。

Melanoma has a poor prognosis for metastatic disease. Despite improvements in its treatment, approximately 50% of patients still develop resistance with widespread relapse and very high mortality. There is growing evidence that a high degree of intra-tumour heterogeneity correlates with poor disease prognosis in patients. Despite large efforts to disentangle the clonal dynamics via genomic and phenotypic studies, our understanding remains rudimentary. This project, therefore, takes a more controlled approach studying how different melanoma clones compete and differ in their vulnerabilities before and during targeted therapy. The ultimate aim is to gain mechanistic insight about how to decrease the fitness of resistant melanoma clones, enabling a more durable therapeutic response. I will generate intra-tumour heterogeneity with a novel approach that stochastically introduces defined clinically relevant oncogenic mutations coupled with unique fluorophores within a tumour that can be tracked in vivo using intravital imaging. The unique fluorophores will enable ex vivo isolation of different clones and downstream mechanistic analysis of changes in signalling and gene expression. Moreover, I will investigate how the tumour microenvironment changes when clones with specific mutations expand. Knowledge acquired by these studies will be exploited to test better treatment strategies in vivo.