A double-switch pHLIP system enables selective enrichment of circulating tumor microenvironment-derived extracellular vesicles.
Zhiyou ZongXinzhuo LiuZhuo YeDingbin LiuPublished in: Proceedings of the National Academy of Sciences of the United States of America (2023)
Circulating tumor microenvironment-derived extracellular vesicles (cTME-EVs) are gaining considerable traction in cancer research and liquid biopsy. However, the study of cTME-EVs is largely limited by the dearth of a general isolation technique to selectively enrich cTME-EVs from biological fluids for downstream analysis. In this work, we broke through this dilemma by presenting a double-switch pH-low insertion peptide (D-S pHLIP) system to exclusively harvest cTME-EVs from the blood serum of tumor mouse models. This D-S pHLIP system consists of a highly sensitive pH-driven conformational switch (p K a ≈ 6.8) that allows specific installation of D-S pHLIP on the EV membranes in TME (pH 6.5 to 6.8) and a unique hook-like switch to "lock" the peptide securely on the cTME-EVs during the systemic circulation. The D-S pHLIP-anchored cTME-EVs were magnetically enriched and then analyzed with high-resolution messenger RNA sequencing, by which more than 18 times the number of TME-related differentially expressed genes and 10 times the number of hub genes were identified, compared with those achieved by the gold-standard ultracentrifugation. This work could revolutionize basic TME research as well as clinical liquid biopsy for cancer.
Keyphrases
- papillary thyroid
- high resolution
- squamous cell
- mouse model
- bioinformatics analysis
- ionic liquid
- single cell
- molecular dynamics
- mass spectrometry
- childhood cancer
- molecular dynamics simulations
- single molecule
- gene expression
- dna methylation
- network analysis
- tandem mass spectrometry
- fluorescent probe
- liquid chromatography
- simultaneous determination