Abstract：
Intercellular heterogeneity occurs widely under both normal physiological environments and abnormal disease-causing conditions. Several attempts to couple spatiotemporal information to cell states in a microenvironment were performed to decipher the cause and effect of heterogeneity. Furthermore, spatiotemporal manipulation can be achieved with the use of photocaged/photoactivatable molecules. Here, we provide a platform to spatiotemporally analyze differential protein expression in neighboring cells by multiple photocaged probes coupled with homemade photomasks. We successfully established intercellular heterogeneity (photoactivable ROS trigger) and mapped the targets (directly ROS-affected cells) and bystanders (surrounding cells), which were further characterized by total proteomic and cysteinomic analysis. Different protein profiles were shown between bystanders and target cells in both total proteome and cysteinome. Our strategy should expand the toolkit of spatiotemporal mapping for elucidating intercellular heterogeneity.
摘要：
细胞间异质性在正常生理环境和异常致病条件下广泛发生。进行了几次尝试将时空信息耦合到微环境中的细胞状态，以破译异质性的原因和影响。此外，时空操纵可以通过使用光老化/光活化分子来实现。这里,我们提供了一个平台，通过多个光老化的探针和自制的光掩模对邻近细胞的差异蛋白表达进行时空分析。我们成功地建立了细胞间异质性（光活化的ROS触发），并绘制了靶标（直接受ROS影响的细胞）和旁观者（周围细胞），通过总蛋白质组学和半胱氨酸组学分析进一步表征。在总蛋白质组和半胱氨酸组中，旁观者和靶细胞之间显示出不同的蛋白质谱。我们的策略应该扩展时空作图的工具包，以阐明细胞间的异质性。
