By Evercyte GmbH
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Evercyte EV and exosome therapeutic support
Evercyte offers development support for innovative exosome research based therapies that rely on the ability of exosome or extracellular vesicles (EVs) to transport proteins, lipids as well as nucleic acids in cellular communication.
Exosomes are small membrane EVs, which are secreted by many cell types that use them to transfer genetic material (biomarkers) from one cell to another. Examples include transfer of microRNAs (miRNAs) to control gene expression of messenger RNAs (mRNAs) to manufacture proteins. They can also transfer DNA, proteins, and lipids.
Exosome markers can thus serve as clinically valuable tools for early diagnosis, prognosis and more targeted exosome therapies. EV and exosome proteomics have already made significant contributions to enhanced understanding of cancer progression and to developing diagnostic biomarkers in carcinogenesis. Most of these innovative exosome-based therapies are still at pre-clinical or early clinical development stages, but have potential to bring exosome treatments to large populations, as demonstrated by the pioneering Pfizer/BioNTech and Moderna vaccines that use mRNA to target the spike proteins of the COVID-19 coronavirus.
Yet exosome research is still in its comparative infancy with data still short on exosome uptake by various cell types and the internal signaling pathways by which EVs elicit cellular response.
Exosome development challenges
Exosomes are produced by all cells with sizes ranging between 30 and 150 nm. Their phospholipid bilayer shells have specific surface markers, while their genetic package contains RNA, DNA and proteins. Exosomes also have potential anti-inflammatory properties through cytokines released during exocytosis.
The main exosome research and development challenges are in assuring consistently high quality and in the accurate characterizations needed to decode the biology and mechanism for exosome therapeutic effect. Current characterization tools include nanoparticle tracking analysis (NTA) for particle sizing and determination of concentrations, electron microscopy, Fluorescence-activated cell sorting (FACS), real-time quantitative Polymerase Chain Reaction (qPCR) and plate-based Enzyme-Linked Immunosorbent Assay (ELISA) for analyzing exosomes and extracted exosome cargo.
Upstream challenges are to assure fully controlled and scalable exosome generation technologies, tissue acquisition, cell isolation, nucleofection and electroporation., along with lentiviral and adeno-associated virus (AAV) transduction methods.
Downstream, challenges include Isolation / purification, formulation implementation for delivery, tangential flow filtration (TFF) for clarification and volume reduction, size-exclusion (SEC) / Ion-exchange for selection of exosomes, compounding of buffer and exosome targeting excipient composition using ultracentrifugation and defining process parameters to achieve final composition of exosomes for delivery.
Exosome-related Evercyte services
Evercyte can establish human highly differentiated cell lines from different organs and tissues for EV production. Principal platforms include:
- Production of recombinant EVs with optimised surface characteristics and cargofor exosome targeting or purification using the Snorkel epitome tagging system.
- Bone marrow derived cells as a rich source of MSCs with a high exosome-based therapeutics
- Fully documented Wharton´s Jelly-derived MSCs established under xeno-free culture conditions
- EV characterization studies including proteomics, lipidomics,, miRNA/mRNA profiling, surface marker profiling, and electron microscopy.
Evercyte’s EV product offerings include hTERT immortalized human adipose-derived mesenchymal stem cell lines with representative differentiation profiles, hTERT immortalized human Wharton´s Jelly and immortalized human bone-marrow derived MSC lines as new EV production systems for EVs.
Research applications include analysis of EV cargo and exosome targeting and generation of recombinant EVs for drug targeting.