Single-Cell Profiling of Fatty Acid Uptake Using Surface-Immobilized Dendrimers

We present a chemical approach to profile fatty acid uptake in single cells. We use azide-modified analogues to probe the fatty acid influx and surface-immobilized dendrimers with dibenzocyclooctyne (DBCO) groups for detection. A competition between the fatty acid probes and BHQ2-azide quencher molecules generates fluorescence signals in a concentration-dependent manner. By integrating this method onto a microfluidics-based multiplex protein analysis platform, we resolved the relationships between fatty acid influx, oncogenic signaling activities, and cell proliferation in single glioblastoma cells. We found that p70S6K and 4EBP1 differentially correlated with fatty acid uptake. We validated that cotargeting p70S6K and fatty acid metabolism synergistically inhibited cell proliferation. Our work provided the first example of studying fatty acid metabolism in the context of protein signaling at single-cell resolution and generated new insights into cancer biology.

Automated summary

This study presents a chemical approach to analyze fatty acid uptake in single cells, which involved using azide-modified analogues and surface-immobilized dendrimers with DBCO groups. By integrating this method into a microfluidics-based multiplex protein analysis platform, the researchers were able to explore the relationships between fatty acid influx, oncogenic signaling activities, and cell proliferation. The study revealed differential correlations between p70S6K and 4EBP1 with fatty acid uptake, and demonstrated synergistic inhibition of cell proliferation through co-targeting p70S6K and fatty acid metabolism. This work provided new insights into cancer biology by studying fatty acid metabolism in the context of protein signaling at single-cell resolution.