Spatiotemporally resolved ex vivo colorectal cancer development in engineered mini-colons (RNA-Seq AKP lines). Spatiotemporally resolved ex vivo colorectal cancer development in engineered mini-colons (RNA-Seq AKP lines)

Here we developed topobiologically complex mini-colons able to undergo tumorigenesis ex vivo by integrating microfabrication, optogenetic, and tissue engineering approaches. With this system, tumorigenic transformation can be spatiotemporally controlled by directing oncogenic activation through blue-light exposure, and emerging colon tumors can be tracked in real-time with single-cell resolution for several weeks without breaking the culture. These induced mini-colons display rich intra- and inter-tumoral diversity and recapitulate key pathophysiological hallmarks displayed by colorectal tumors in vivo. By fine-tuning cell-intrinsic and extrinsic parameters, mini-colons can be leveraged to unveil tumorigenic determinants, including dietary patterns, microbiota-derived metabolites, and pharmacological therapies. Overall design: Gene expression profiling analysis of bulk RNA-seq data from colon organoid lines derived from: healthy colon (control), OptoCre-induced AKP organoids (organoid AKP), OptoCre-induced AKP mini-colons (mini-colon AKP), in vivo colon tumors (in vivo AKP).

本研究通过整合微加工、光遗传学与组织工程技术，构建了可在体外发生肿瘤发生进程的、具有拓扑生物学复杂性的微型结肠组织（mini-colons）。借助该模型体系，可通过蓝光照射诱导致癌激活，实现肿瘤转化的时空精准调控；且可在不破坏培养体系的前提下，以单细胞分辨率实时追踪新生结肠肿瘤长达数周之久。此类诱导得到的微型结肠组织展现出丰富的瘤内与瘤间异质性，并可复现体内结直肠肿瘤的关键病理生理学特征。通过微调细胞内在与外在参数，该微型结肠组织模型可用于揭示肿瘤发生的核心决定因素，涵盖饮食模式、菌群衍生代谢物以及药物治疗等研究方向。实验整体设计：对源自以下样本的结肠类器官系的批量RNA测序（bulk RNA-seq）数据开展基因表达谱分析：健康结肠（对照组）、OptoCre诱导的AKP类器官（类器官AKP）、OptoCre诱导的AKP微型结肠组织（微型结肠AKP）以及体内结肠肿瘤（体内AKP）。