Deconstructing tissue-specific effects of genes and variants on proliferation is critical to understanding cellular transformation and systematically selecting cancer therapeutics. This requires scalable methods for multiplexed genetic screens tracking fitness across time, across lineages, and in a suitable niche, since physiological cues influence functional differences. Towards this, we present an approach, coupling single-cell cancer driver screens in teratomas with hit enrichment by serial teratoma reinjection, to simultaneously screen drivers across multiple lineages in vivo. Using this system, we analyzed population shifts and lineage-specific enrichment for 51 cancer associated genes and variants, profiling over 100,000 cells spanning over 20 lineages, across two rounds of serial reinjection. We confirmed that c-MYC alone or combined with myristoylated AKT1 potently drives proliferation in progenitor neural lineages, demonstrating signatures of malignancy. Additionally, mutant MEK1^S218D/S222D provides a proliferative advantage in mesenchymal lineages like fibroblasts. Our method provides a powerful platform for multi-lineage longitudinal study of oncogenesis.

解构基因和变异对增殖的组织特异性影响对于理解细胞转化和系统地选择癌症治疗方法至关重要。这需要可扩展的多重遗传筛选方法来跟踪跨时间、跨谱系和在合适的利基中的健康状况，因为生理线索会影响功能差异。为此，我们提出了一种方法，将畸胎瘤中的单细胞癌症驱动筛选与通过连续畸胎瘤回注进行的命中富集相结合，以在体内同时筛选跨多个谱系的驱动。使用该系统，我们分析了 51 个癌症相关基因和变异的群体变化和谱系特异性富集，在两轮连续再注射中对跨越 20 多个谱系的 100,000 多个细胞进行了分析。我们证实，c-MYC单独使用或与肉豆蔻酰化AKT1联合使用可有效促进祖神经谱系的增殖，从而展现出恶性肿瘤的特征。此外，突变型MEK1 ^S218D/S222D在间充质谱系（如成纤维细胞）中提供了增殖优势。我们的方法为肿瘤发生的多谱系纵向研究提供了一个强大的平台。