Detectable Lipidomes and Metabolomes by Different Plasma Exosome Isolation Methods in Healthy Controls and Patients...

Area of Science:

• Biochemistry
• Molecular Biology
• Oncology

Background:

• Circulating exosomes in blood hold potential as cancer biomarkers.
• Exosome heterogeneity means isolation methods can yield distinct molecular profiles.
• Evaluating isolation method impact is key for reliable biomarker discovery.

Purpose of the Study:

• To assess how plasma exosome isolation techniques affect multi-omic profiles in non-small cell lung cancer (NSCLC) and castration-resistant prostate cancer (CRPC).
• To develop an algorithm for quantifying exosome enrichment.
• To compare size exclusion chromatography (SEC), lectin binding, and TIM-4 binding methods.

Main Methods:

• Plasma exosomes isolated from CRPC, NSCLC patients, and controls using SEC, lectin, and TIM-4 binding.
• Molecular profiling via mass spectrometry.
• Enrichment analysis using MetaboAnalyst and an exosome enrichment index (EEI).

Main Results:

• SEC yielded the highest number of detected lipids (949) and metabolites (191).
• SEC identified significant lipid differences in NSCLC patients compared to controls.
• Lectin and TIM-4 binding methods showed fewer detected molecules and no significant differences between patient groups.

Conclusions:

• Size exclusion chromatography (SEC) captures more heterogeneous exosome populations, enhancing biomarker discovery potential.
• Lectin and TIM-4 binding methods preferentially isolate specific exosome subpopulations.
• The choice of exosome isolation method significantly influences cancer biomarker discovery outcomes.