Department of Biological Sciences, Columbia University, New York City, NY 10027, USA.
Department of Biology, Barnard College, New York, NY 10027, USA; Institute for Cancer Genetics, Department of Genetics and Development, Columbia University Irving Medical Center, New York, NY 10032, USA; Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, NY 10032, USA.
Cell Rep Methods. 2024 May 20;4(5):100760. doi: 10.1016/j.crmeth.2024.100760. Epub 2024 Apr 26.
The role of protein turnover in pancreatic ductal adenocarcinoma (PDA) metastasis has not been previously investigated. We introduce dynamic stable-isotope labeling of organoids (dSILO): a dynamic SILAC derivative that combines a pulse of isotopically labeled amino acids with isobaric tandem mass-tag (TMT) labeling to measure proteome-wide protein turnover rates in organoids. We applied it to a PDA model and discovered that metastatic organoids exhibit an accelerated global proteome turnover compared to primary tumor organoids. Globally, most turnover changes are not reflected at the level of protein abundance. Interestingly, the group of proteins that show the highest turnover increase in metastatic PDA compared to tumor is involved in mitochondrial respiration. This indicates that metastatic PDA may adopt alternative respiratory chain functionality that is controlled by the rate at which proteins are turned over. Collectively, our analysis of proteome turnover in PDA organoids offers insights into the mechanisms underlying PDA metastasis.
蛋白质周转在胰腺导管腺癌（PDA）转移中的作用以前尚未被研究过。我们引入了类器官的动态稳定同位素标记（dSILO）：一种动态 SILAC 衍生物，它结合了脉冲的同位素标记氨基酸和同位素质谱标签（TMT）标记，以测量类器官中蛋白质组范围的蛋白质周转速率。我们将其应用于 PDA 模型中，发现与原发性肿瘤类器官相比，转移性类器官表现出加速的全局蛋白质组周转。在全局范围内，大多数周转率变化并未反映在蛋白质丰度水平上。有趣的是，在转移性 PDA 中与肿瘤相比，蛋白质周转增加最多的蛋白质组参与线粒体呼吸。这表明转移性 PDA 可能采用由蛋白质周转率控制的替代呼吸链功能。总的来说，我们对 PDA 类器官中蛋白质组周转的分析为 PDA 转移的机制提供了新的见解。
