Integrative single-cell metabolomics and phenotypic profiling reveals metabolic heterogeneity of cellular oxidation and senescence
Ziyi Wang, Si-Yuan Ge, Tiepeng Liao, Man Yuan, Wenwei Qian, Qi Chen, Wei Liang, Xiawei Cheng, Qinghua Zhou, Zhenyu Ju, Hongying Zhu, Wei Xiong
Abstract
Emerging evidence has unveiled heterogeneity in phenotypic and transcriptional alterations at the single-cell level during oxidative stress and senescence. Despite the pivotal roles of cellular metabolism, a comprehensive elucidation of metabolomic heterogeneity in cells and its connection with cellular oxidative and senescent status remains elusive. By integrating single-cell live imaging with mass spectrometry (SCLIMS), we establish a cross-modality technique capturing both metabolome and oxidative level in individual cells. The SCLIMS demonstrates substantial metabolomic heterogeneity among cells with diverse oxidative levels. Furthermore, the single-cell metabolome predicted heterogeneous states of cells. Remarkably, the pre-existing metabolomic heterogeneity determines the divergent cellular fate upon oxidative insult. Supplementation of key metabolites screened by SCLIMS resulted in a reduction in cellular oxidative levels and an extension of C. elegans lifespan. Altogether, SCLIMS represents a potent tool for integrative metabolomics and phenotypic profiling at the single-cell level, offering innovative approaches to investigate metabolic heterogeneity in cellular processes. Integrated analysis of metabolome and oxidative stress at single-cell level is challenging. Here, the authors develop SCLIMS, enabling simultaneous profiling of metabolome and oxidative stress levels and discoveries of key metabolites regulating oxidative stress, senescence, and healthy aging.