Delivery of nucleic acids, proteins and small molecules in vitreous vesicular bodies
The present invention relates to compositions of aqueous humor and/or vitreous humor derived extracellular vesicles and their use for the delivery of therapeutic agents to ocular tissues for the treatment of ophthalmic diseases. Further disclosed are methods of making the compositions. Methods of treating and diagnosing an ocular condition are also disclosed.
1 . A composition comprising:
one or more isolated, aqueous humor and/or vitreous humor extracellular vesicle bodies, wherein said extracellular vesicle bodies are modified in vitro to contain one or more exogenous agents, wherein endogenous contents of said extracellular vesicle bodies are removed, wherein said extracellular vesicle bodies are from 100 to 6,000 nanometers in diameter, wherein the composition is at least 75% free of cell or cellular debris, and wherein said extracellular vesicle bodies are modified to display a eukaryotic cell-specific targeting molecule on the vesicle surface so that the composition mediates uptake of the one or more exogenous agents by the eukaryotic cells, wherein the eukaryotic cells are selected from the group consisting of human skin cells, human retinal cells, human ciliary cells, and human corneal cells.
2 . The composition of claim 1 , wherein the one or more exogenous agents is selected from the group consisting of a nucleic acid molecule, a protein or polypeptide, a small molecule, a hormone, and any combination thereof.
3 . The composition of claim 1 , wherein the one or more exogenous agents comprise one or more nucleic acid molecules selected from the group consisting of a ribonucleic acid, a small RNA molecule, a complementary RNA, a non-coding RNA molecule, a siRNA, a pi-RNA molecule, a micro-RNA molecule, a sno-RNA molecule, a long non-coding RNA molecule, a messenger RNA molecule, a ribosomal RNA molecule, an antisense nucleic acid molecule, a Locked Nucleic Acid (LNA), an antagomir, a CRISPR/Cas gene editing RNA, a trans-activating crRNA (tracrRNA), a short synthetic RNA composed of a “scaffold” sequence (gRNA), a Small Cajal body-specific RNAs (scaRNA), a natural cis-antisense siRNAs (cis-nat-siRNAs), a trans-acting siRNA (tasiRNA), a repeat associated small interfering RNA (rasiRNA), a 7SK RNA, a transfer-messenger RNA (tmRNA), a transfer RNA (tRNA), a 7SL RNA, a signal recognition particle RNA (SRP), and any combination thereof.
4 . The composition of claim 1 , wherein the one or more exogenous agents comprise one or more of a small deoxy-ribonucleic acid (DNA) molecule, a cDNA molecule, an oligonucleotide, a locked Nucleic Acid (LNA), a deoxyribonucleic acid aptamer, a deoxyribonucleic acidzyme, and any combination thereof.
5 . The composition of claim 1 , wherein the one or more exogenous agents are naked or packaged in a viral vector, a bacterial vector, a plasmid vector, or any combination thereof.
6 . The composition of claim 1 , wherein the composition comprises a pharmaceutically acceptable carrier.
7 . The composition of claim 1 , wherein said composition is formulated in a slow or sustained release material.
8 . The composition of claim 1 , wherein the exogenous agent is a therapeutic agent.
9 . The composition of claim 1 , wherein the exogenous agent is a diagnostic agent.
10 . The composition of claim 1 , wherein the one or more extracellular vesicle bodies are isolated from mammalian vitreous humor and/or aqueous humor.
11 . The composition of claim 1 , wherein the one or more extracellular vesicle bodies are isolated from human vitreous humor and/or aqueous humor.