Extracellular vesicles as vehicles for small non-coding RNA therapeutics: standardization challenges for clinical...

Area of Science:

• Biotechnology
• Molecular Biology
• Drug Delivery

Background:

• Extracellular vesicles (EVs) are emerging as potent carriers for small non-coding RNAs (sncRNAs).
• Clinical translation of EV-based RNA therapeutics is impeded by non-standardized manufacturing, loading, and dosing protocols.
• A comprehensive analysis is needed to identify trends and challenges in the field.

Purpose of the Study:

• To review and analyze the current landscape of sncRNA-loaded EVs in preclinical and clinical research.
• To identify key trends, discrepancies, and challenges in EV production, RNA loading, and dosing strategies.
• To provide insights for the future development of EV-based RNA therapeutics.

Main Methods:

• Systematic literature search of PubMed and Google Scholar for articles on cell-derived EVs loaded with sncRNA.
• Quantitative analysis of EV source, surface modifications, RNA type, loading methods, and dosages from 74 selected studies.
• Identification of trends and discrepancies in manufacturing and application protocols.

Main Results:

• Stem or immortalized cell-derived EVs are predominantly used, with electroporation and donor cell transfection as common loading methods.
• Preclinical studies primarily use protein content for EV loading and dosage, with limited use of particle number.
• Significant variability in measurement units and lack of standardized guidelines complicate result comparison across studies.

Conclusions:

• Standardized guidelines for EV production, RNA loading, and dosing are critically needed for therapeutic development.
• Reliable dosing strategies are essential for evaluating the therapeutic potential of EVs and ensuring clinical translatability.
• Establishing universal quantification methods and reproducible protocols is paramount for advancing EV-based RNA delivery platforms.