Human CAP cells represent a novel source for functional, miRNA-loaded exosome production

Abstract

Exosomes represent a promising delivery tool for nucleic acid-based pharmaceuticals. They are highly suitable for transporting therapeutic miRNAs to tumor cells, due to their natural membrane components. Further, exosomes are capable of effectively protecting nucleic acids against ribonucleases and enable the delivery of their content through cell membranes. However, no suitable production host for miRNA containing exosomes of non-tumorigenic origin has yet been identified. In this study we engineered an immortalised human amniocyte cell line (CAP ® cells), whose exosomes were enriched and characterised. The cell line modifications not only enabled the production of GFP-labelled but also pro-apoptotic miRNA containing exosomes without negative influence on host cell growth. Furthermore, we demonstrated that pro-apoptotic miRNA containing CAP exosomes are taken up by ovarian cancer cells. Strikingly, delivery of functional exosomal miRNA led to downregulation of several reported target genes in the treated tumor cells. In summary, we revealed CAP cells of non-tumorigenic origin as a novel and efficient exosome production host with the potential to produce functional miRNA-loaded exosomes.

Towards analyzing the potential of exosomes to deliver microRNA therapeutics

“…The efficient delivery of Alexa Fluor 488‐conjugated probe to cancer cells utilizing exosomes from non‐tumorigenic cell lines paves the way towards the standardization of techniques that will allow the preclinical and clinical development of exosome‐based “bio‐shuttles” of molecular therapeutic cargo delivery. To this end, it is noteworthy that a recent research effort indicates that the engineering of an immortalized human amniocyte cell line can serve as a novel and efficient exosome production host with the potential to produce functional miRNA‐loaded exosomes (Zeh et al, 2019).…”

Therapeutic miRNA-Enriched Extracellular Vesicles: Current Approaches and Future Prospects

“…Both contained motifs in 3p strand, which helped them to load in EVs. Further, they can unload their cargo in ovarian cancer cells (SKOV3), thus, down-regulating mRNA targets [ 20 ]. Additionally, the immortalized cell line from tumor cells can provide high yield of EVs, but cannot be used for therapeutic purpose due to safety issues [ 19 ].…”

Exploring the molecular content of CHO exosomes during bioprocessing

“…Moreover, exosomes are considered potential drug delivery vehicle, due to their specific tropism, proposedly weak immunogenicity and the ability to cross the blood-brain-barrier (Liu and Su 2019 ). Therefore, tailored exosomes packed up with small molecules (Lamichhane and Jay 2018 ), miRNA (Zeh et al 2019 ) or Fab fragments attached to their surface (Longatti et al 2018 ) have been studied.…”