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Background <p>Mechanisms governing non-vesicular extracellular particle (NVEP) biogenesis and compositional diversity remain incompletely characterized.</p> Method <p>This study integrates multiparametric profiling to define NVEPs. NVEPs were isolated by differential ultracentrifugation and characterized using complementary biochemical, lipidomic, and imaging approaches, including protein-to-lipid ratio analysis, lipid profiling, and advanced microscopy.</p> Result <p>Comprehensive characterization demonstrated that NVEPs exhibit significantly elevated protein-to-lipid ratios compared to extracellular vesicles (EVs) subtypes. Lipidomic profiling revealed pronounced enrichment in cholesteryl esters and triacylglycerols, with distinct carbon chain length signatures differentiating NVEPs from small EVs. Super-resolution microscopy confirmed marker heterogeneity across populations, with NVEPs showing specific enrichment of Arf6 and CD63.</p> Conclusion <p>These findings provide a robust compositional framework to distinguish non-vesicular extracellular particles from canonical EVs and underscore their emerging significance as mediators of intercellular communication and potential vehicles for targeted delivery.</p>

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Cholesteryl esters and high protein-to-lipid ratios distinguish non-vesicular extracellular particles from extracellular vesicles

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月17日

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  • Sayam Ghosal,
  • Rita Leporati,
  • Árpád Varga,
  • Petra Susanszki,
  • Nóra Fekete,
  • Tamás László,
  • Tünde Bárkai,
  • Fülöp Károly Grébecz,
  • Delaram Khamari,
  • Tárek Zoltán Magyar,
  • Marcus Hoering,
  • Krisztina V Vukman,
  • Bernadett R Bodnár,
  • Brachyahu M Kestecher,
  • Mohamed A Fattah,
  • Csaba Bödör,
  • József Maléth,
  • Gerhard Liebisch,
  • Evelyn Orsó,
  • Edit I Buzas,
  • Xabier Osteikoetxea

摘要

Background

Mechanisms governing non-vesicular extracellular particle (NVEP) biogenesis and compositional diversity remain incompletely characterized.

Method

This study integrates multiparametric profiling to define NVEPs. NVEPs were isolated by differential ultracentrifugation and characterized using complementary biochemical, lipidomic, and imaging approaches, including protein-to-lipid ratio analysis, lipid profiling, and advanced microscopy.

Result

Comprehensive characterization demonstrated that NVEPs exhibit significantly elevated protein-to-lipid ratios compared to extracellular vesicles (EVs) subtypes. Lipidomic profiling revealed pronounced enrichment in cholesteryl esters and triacylglycerols, with distinct carbon chain length signatures differentiating NVEPs from small EVs. Super-resolution microscopy confirmed marker heterogeneity across populations, with NVEPs showing specific enrichment of Arf6 and CD63.

Conclusion

These findings provide a robust compositional framework to distinguish non-vesicular extracellular particles from canonical EVs and underscore their emerging significance as mediators of intercellular communication and potential vehicles for targeted delivery.