The Proteo-Transcriptome of Extracellular Vesicles and Particles Is Largely Preserved After Cryopreservation

Area of Science:

• Biochemistry
• Molecular Biology
• Cancer Research

Background:

• Extracellular vesicles and particles (EVPs) are vital in tumorigenesis, metastasis, and immunomodulation.
• Understanding EVP biology requires reliable tissue sources, but the impact of cryopreservation on EVPs is not fully understood.

Purpose of the Study:

• To investigate the proteo-transcriptomic differences in EVP cargo between fresh and frozen human tissues.
• To assess the feasibility of using cryopreserved tissues for EVP research.

Main Methods:

• EVPs were extracted from fresh and frozen tumor and adjacent normal human tissues using ultrafiltration and size-exclusion chromatography.
• Protein and RNA content were profiled using spectroscopy.
• Olink technology and mixed-effect models were employed to analyze protein content differences.

Main Results:

• Cryopreservation demonstrated minimal effects on EVP protein concentration and particle size.
• Proteo-transcriptomic cargo of EVPs remained largely consistent between fresh and frozen samples.
• No significant alterations were observed in EVP composition due to cryopreservation.

Conclusions:

• Frozen tissues are suitable for extracellular vesicle and particle research.
• Cryopreservation does not significantly affect EVP cargo, supporting the use of large frozen biobanks.
• This research overcomes a major hurdle in human sample research, enhancing accessibility and reducing reliance on fresh tissues.