RNA nanoparticles and method of use thereof
The presently-disclosed subject matter relates to an artificial RNA nanostructure and method of use thereof. In particular, the presently-disclosed subject matter relates to RNA nanoparticles and RNA dendrimers, and methods of disease diagnosis and treatments using the RNA nanostructure and RNA dendrimers.
1. A method of diagnosing cancer in a subject, the method comprising administering to the subject an RNA dendrimer molecule, wherein the RNA dendrimer molecule comprises (a) a central core multi-branched RNA junction motif, wherein the central core motif comprises a plurality of branches, wherein a branch comprises at least one RNA oligonucleotide, and wherein the central core motif comprises at least one RNA oligonucleotide having at least 90% identity to SEQ ID NO: 12 (SquareE); and (b) an outer surface multi-branched RNA junction motif comprising at least one repeating multi-branched RNA junction motif unit, wherein the repeating unit comprises a plurality of branches, and wherein a branch comprises at least one RNA oligonucleotide, wherein the RNA dendrimer molecule comprises at least one imaging module, wherein the RNA dendrimer molecule comprises a folate receptor targeting module, wherein the cancer comprises a cancer that overexpresses folate receptor, and diagnosing the subject as having cancer if the RNA dendrimer molecule is detected in the subject.
2. The method of claim 1 , wherein the RNA dendrimer molecule further comprises at least one targeting module, at least one therapeutic module, or a combination thereof.
3. The method of claim 1 , wherein the plurality of branches comprising the central core multi-branched RNA junction motif comprises a three-branched RNA junction motif, or four-branched RNA junction motif, a six-branched RNA junction motif, or an eight-branched RNA junction motif.
4. The method of claim 1 , wherein the plurality of branches comprising the repeating multi-branched RNA junction motif unit comprises a three-branched RNA junction motif, or a four-branched RNA junction motif, a six-branched RNA junction motif, and an eight-branched RNA junction motif.
5. The method of claim 3 or 4 , wherein the three-branched RNA junction motif comprises an a3WJ RNA module having at least 90% identity to nucleotide sequence 5′-UUG CCA UGU GUA UGU GGG-3′ (SEQ ID NO: 1); a b3WJ RNA module having at least 90% identity to nucleotide sequence 5′-CCC ACA UAC UUU GUU GAUCC-3′ (SEQ ID NO: 2); or a c3WJ RNA module having at least 90% identity to nucleotide sequence 5′-GGA UCA AUC AUG GCA A-3′ (SEQ ID NO: 3), or a combination thereof.
6. The method of claim 3 or 4 , wherein the central core multi-branched RNA junction motif is a polygon shaped architecture comprising a three-branched RNA junction motif at each corner.
7. The method of claim 1 , wherein the imaging module comprises a fluorescent dye, a radionuclide, a contrast agent, a reporter imaging molecule, a reference imaging molecule, or any combination thereof.
8. The method of claim 7 , wherein the reference imaging module comprises a reference dye and a quencher.
9. The method of claim 7 , wherein the contrast agent comprises at least one MRI contrast agent, at least one gadolinium contrast agent, or both.
10. The method of claim 2 , wherein the targeting module comprises a cancer targeting module.
11. The method of claim 10 , wherein the cancer targeting module is a ligand that binds to at least one cancer cell surface marker.
12. The method of claim 10 , wherein the targeting module comprises a chemical ligand or RNA aptamer.
13. The method of claim 11 , wherein the ligand comprises folate.
14. The method of claim 11 , wherein the cell surface marker comprises a folate receptor.
15. The method of claim 11 , wherein the ligand binds to an epithelial cell adhesion molecule (EpCAM).
16. The method of claim 11 , wherein the cell surface marker comprises an epithelial cell adhesion molecule (EpCAM).
17. The method of claim 12 , wherein the aptamer is an EpCAM RNA aptamer.
18. The method of claim 2 , wherein the therapeutic module comprises a siRNA, a miRNA, an anti-mRNA, a ribozyme RNA, an antisense RNA, a chemotherapeutic drug, a cytotoxic drug, a riboswitch, or an endosome disrupting agent.
19. The method of claim 18 , wherein the cytotoxic drug comprises doxorubicin, paclitaxel, paclitaxel derivatives and analogues, cytochalasin D, rapamycin, rapamycin derivatives and analogues, camptothecin, dexamethasone, and 5-fluorouracil, a quinazolinone derivative, metallic silver, tranilast, everolimus and/or related compounds.
20. The method of claim 1 , wherein the RNA dendrimer molecule further comprises an oligonucleotide having at least 90% identity to nucleotide sequences of at least one of:
3WJ-a: UUG CCA UGU GUA UGU GGG (SEQ ID NO: 1)
3WJ-b: CCC ACA UAC UUU GUU GAU CC (SEQ ID NO: 2)
3WJ-c: GGA UCA AUC AUG GCA A (SEQ ID NO: 3)
3WJ-a_rev: GGG UGU AUG UGU ACC GUU (SEQ ID NO: 4)
3WJ-b_rev: CCU AGU UGU UUC AUA CAC CC (SEQ ID NO: 5)
3WJ-c_rev: AAC GGU ACU AAC UAG G (SEQ ID NO: 6)
3WJ-c-folate: GGA UCA AUC AUG GCA A—folate (SEQ ID NO: 7)
SquareA: GGG AGC CGU CAA UCA UGG CAA GUG UCC GCC AUA CUU UGU UGC ACG CAC (SEQ ID NO: 8)
SquareB: GGG AGC GUG CAA UCA UGG CAA GCG CAU CGC AUA CUU UGU UGC GAC CUA (SEQ ID NO: 9)
SquareC: GGG AGG UCG CAA UCA UGG CAA CGA UAG AGC AUA CUU UGU UGG CUG GAG (SEQ ID NO: 10)
SquareD: GGG ACC AGC CAA UCA UGG CAA UAU ACA CGC AUA CUU UGU UGA CGG CGG (SEQ ID NO: 11)
b-SquareA: GGC CCA CAU ACU UUG UUG AUC CAU GGU GCG UAG GGU CGU CAA UCA UGG CAA GUG UCC GCC AUA CUU UGU UGC ACU CCC UUG CUC AUC A (SEQ ID NO: 13)
b-SquareB: GGC CCA CAU ACU UUG UUG AUC CUG AUG AGC AAG GGA GUG CAA UCA UGG CAA GCG UAU CGC AUA CUU UGU UGA GAA CCC UAU GUG ACU U (SEQ ID NO: 14)
b-SquareC: GGC CCA CAU ACU UUG UUG AUC CAA GUC ACA UAG GGU UCG CAA UCA UGG CAA CGA UAG AGC AUA CUU UGU UGG AGU CCC UUA GAG UAG A (SEQ ID NO: 15)
b-SquareD: GGC CCA CAU ACU UUG UUG AUC CUC UAC UCU AAG GGA CUC CAA UCA UGG CAA UAU ACA CGC AUA CUU UGU UGA CGA CCC UAC GCA CCA U (SEQ ID NO: 16)
3WJ-a-b_rev: UUG CCA UGU GUA UGU GGG CCU AGU UGU UUC AUA CAC CC (SEQ ID NO: 17)
3WJ-c-b_rev: GGA UCA AUC AUG GCA A CCU AGU UGU UUC AUA CAC CC (SEQ ID NO: 18)
3WJ-b-a_rev: CCC ACA UAC UUU GUU GAU CC GGG UGU AUG UGU ACC GUU (SEQ ID NO: 19) or
3WJ-b-c_rev: CCC ACA UAC UUU GUU GAU CC AAC GGU ACU AAC UAG G (SEQ ID NO: 20).
21. The method of claim 1 , wherein RNA dendrimer is administered as a composition comprising a therapeutically effective amount of the RNA dendrimer molecule.
22. The method of claim 1 , wherein the central core motif further comprises an RNA oligonucleotide having at least 90% identity to at least one of SEQ ID NO: 8 (SquareA), SEQ ID NO: 9 (SquareB), SEQ ID NO: 10 (SquareC), SEQ ID NO: 11 (SquareD), SEQ ID NO: 13 (b-SquareA), SEQ ID NO: 14 (b-SquareB), SEQ ID NO: 15 (b-SquareC), or SEQ ID NO: 9 (b-SquareD).
23. The method of claim 21 , wherein the composition further comprises a pharmaceutically acceptable carrier.
24. The method of claim 1 , wherein the subject is a mammal or a non-mammal vertebrate.
25. The method of claim 1 , wherein the subject is a human.
26. The method of claim 1 , wherein the cancer is a colon cancer.