RNA ligand-displaying exosomes for specific delivery of therapeutics to cell by RNA nanotechnology
Disclosed herein are compositions comprising extracellular vesicles, such as exosomes, displaying an RNA nanoparticle on its surface. The RNA nanoparticle can target the extracellular vesicle to a given cell via a targeting moiety. The extracellular vesicle can also comprise a functional moiety, which can be used in treatment or diagnostics.
1. A composition comprising an RNA nanoparticle anchored on the surface of an extracellular vesicle membrane,
wherein the nanoparticle is assembled from one or more ribonucleic acid strands duplexed together to form a secondary structure with three or more projecting stem loops,
wherein at one of the three or more projecting stem loops is conjugated to a cholesterol moiety
wherein at one of the three or more projecting stem loops comprises one or more functional moieties, and
wherein at least one of the three or more projecting stem loops physically blocks encapsulation of the nanoparticle into the extracellular vesicle.
2. The composition of claim 1 , wherein at least one of the three or more ribonucleic acid strands comprise a pRNA-3WJ core.
3. The composition of claim 1 , wherein the RNA nanoparticle is assembled from three ribonucleic acid strands comprising the nucleic acid sequence SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3.
4. The composition of claim 1 , wherein one or more of the functional moieties comprises a targeting moiety.
5. The composition of claim 4 , wherein the targeting moiety directs the exosome to a cell of interest.
6. The composition of claim 4 , wherein the targeting moiety is selected from an RNA aptamer, modified RNA aptamer, DNA aptamer, modified DNA aptamer, and chemical ligand.
7. The composition of claim 1 , wherein one or more of the functional moieties comprises a therapeutic moiety or a diagnostic moiety.
8. The composition of claim 7 , wherein the therapeutic moiety or a diagnostic moiety comprises an RNA aptamer, a ribozyme, siRNA, protein-binding RNA aptamer, or small molecule.
9. The composition of claim 1 , wherein the extracellular vesicle comprises an exosome.
10. A method of targeting an extracellular vesicle to a cell of interest comprising: contacting the cell with a composition comprising an extracellular vesicle displaying an RNA nanoparticle on its surface,
wherein the nanoparticle is assembled from one or more ribonucleic acid strands duplexed together to form a secondary structure with three or more projecting stem loops,
wherein at one of the three or more projecting stem loops is conjugated to a cholesterol moiety,
wherein at least one of the three or more projecting stem loops physically blocks encapsulation of the nanoparticle into the extracellular vesicle, and
wherein at one of the three or more projecting stem loops comprises at least one targeting moiety that directs the extracellular vesicle to the cell of interest.
11. The method of claim 10 , wherein the cell is in a subject.
12. The method of claim 10 , wherein the cell is a cancer cell.
13. The method of claim 10 , wherein the RNA nanoparticle further comprises a functional moiety.
14. A method of treating disease in a subject, comprising administering to the subject composition comprising an extracellular vesicle displaying an RNA nanoparticle on its surface,
wherein the nanoparticle is assembled from one or more ribonucleic acid strands duplexed together to form a secondary structure with three or more projecting stem loops,
wherein at one of the three or more projecting stem loops is conjugated to a cholesterol moiety,
wherein at least one of the three or more projecting stem loops physically blocks encapsulation of the nanoparticle into the extracellular vesicle, and
wherein at one of the three or more projecting stem loops comprises one or more functional moieties capable of treating the disease in the subject.
15. The method of claim 14 , wherein the disease is an infection.
16. The method of claim 14 , wherein the disease is cancer.
17. A method of imaging a cell, the method comprising contacting the cell with a composition comprising an extracellular vesicle displaying an RNA nanoparticle on its surface,
wherein the nanoparticle is assembled from one or more ribonucleic acid strands duplexed together to form a secondary structure with three or more projecting stem loops,
wherein at one of the three or more projecting stem loops is conjugated to a cholesterol moiety
wherein at least one of the three or more projecting stem loops physically blocks encapsulation of the nanoparticle into the extracellular vesicle, and
wherein at one of the three or more projecting stem loops comprises one or more diagnostic moieties.
18. The method of claim 17 , wherein the cell is in a subject.