Method for producing therapeutic exosomes from nanoelectroporation based cell transfection
T2018-015 A method for producing therapeutic extracellular vesicles that contain functional mRNAs, miRNAs and shRNAs, for non-endocytic deliver of DNA for applications in cancer treatment and regenerative medicine.
Numerous cell types secrete extracellular vesicles (EVs), including exosomes and microvesicles. Exosomes are nano-vesicles (40–150 nm), and contain both coding and non-coding RNAs and their fragments, DNA fragments, proteins, and other cell related biomolecules. EVs and their biomolecule contents have been proposed as effective biomarkers for disease diagnosis and can play major roles in cell-cell communications in tumor microenvironment and circulation. EVs loaded with functional RNAs and proteins have also been suggested as drugs and drug carriers for therapeutic applications. To deliver specific nucleic acids and/or proteins to target tissues or cell types in vitro and in vivo requires methods that can produce EVs with either endogenous or exogenous therapeutic cargos. The current approach for insertion RNA plasmids into pre-existed exosomes faces many significant limitations in efficiency and low yield. There is a great need for new methods for transfection of EVs containing RNA therapeutics.
Researchers at The Ohio State University, led Dr. L. James Lee, have developed a method for producing therapeutic EVs that contain functional mRNAs, miRNAs and shRNAs, for non-endocytic deliver of DNA plasmids. This was accomplished through the use of nanochannel electroporation (NEP) biochip to transfect donor cells with DNA plasmids. This technology demonstrates that NEP can produce a large number of therapeutic exosomes containing high copies of functional mRNA and microRNA targets, not achievable by previously developed post-insertion methods.
- Cancer Therapy
- Wound Treatment
- Regenerative Tissue and Organ Therapy
- This technology could be deployed via non-endocytic methods such as gene gun, micro-nano injection as they provide proper cell stimulation and fast vector delivery
- Non-endocytic delivery of DNA plasmids/vectors leads to fast transcription of RNAs and translation of proteins within cytoplasm, allowing those functional biomolecules to be encapsulated in the vesicles endogenously before they are secreted out from donor cells as EVs