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

黑色素瘤的转移性疾病预后较差。尽管在治疗上有所改进，但大约50%的患者仍然表现出耐药性，普遍复发，死亡率非常高。越来越多的证据表明，肿瘤内高度的异质性与患者较差的疾病预后相关。尽管我们通过基因组和表型研究做出了巨大努力来解开克隆动力学，但我们的理解仍处于初级阶段。因此，这个项目采取了一种更受控制的方法，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，以及它们的脆弱性的差异。最终目的是获得关于如何降低耐药黑色素瘤克隆的适合性的机械性见解，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的异质性，这种方法随机引入定义的临床相关的致癌基因突变，并在肿瘤内结合独特的荧光团，可以使用活体成像在体内进行跟踪。独特的荧光团将使不同克隆的体外分离和下游信号和基因表达变化的机制分析成为可能。此外，我将研究当具有特定突变的克隆扩大时，肿瘤微环境是如何变化的。通过这些研究获得的知识将被用来在体内测试更好的治疗策略。

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.