Encapsulated nanoparticles for nucleic acid delivery
Methods and compositions for delivering agents (e.g., gene silencing agents) and molecules to cells using yeast cell wall particles are presented herein. Embodiments of the invention are particularly useful for the delivery of nucleic acids (e.g., siRNAs) to cells.
1. A method of reducing or inhibiting inflammation in a subject having an inflammatory disease or disorder comprising,
administering to the subject a siRNA-delivery system, comprising yeast cell wall particles (YCWPs), wherein the YCWPs individually comprise a yeast cell wall particle (YCWP) exterior and a multilayered nanoparticle interior, wherein the multilayered nanoparticle interior comprises a cationic nucleic acid core, coated with at least one siRNA layer, wherein the siRNA layer is coated with at least one cationic trapping layer, wherein the multilayered nanoparticle interior is produced by first introducing into the YCWP exterior, components to form the cationic nucleic acid core such that the core forms inside the YCWP exterior, followed by introducing siRNA such that the siRNA layer forms on the surface of the core, followed by introducing a trapping molecule such that the trapping layer forms on the siRNA layer, and wherein the siRNA targets a gene involved in inflammation,
wherein the siRNA is present at a ratio of about 0.1×10 −7 to 100×10- 7 pmol siRNA per YCWP,
wherein the siRNA-delivery system is administered under conditions facilitating delivery of the siRNA to phagocytic cells of the subject, such that inflammation is reduced or inhibited in the subject having the inflammatory disease or disorder.
2. The method of claim 1 , wherein the conditions facilitate phagocytosis of the siRNA delivery system, such that delivery of the siRNA occurs.
3. The method of claim 1 , wherein the phagocytic cells are selected from the group consisting of a monocyte, monocytic cells, neutrophil cells, dendritic cells and macrophage cells.
4. The method of claim 1 , wherein the phagocytic cells are macrophage cells.
5. The method of claim 1 , wherein the inflammatory disease or disorder is a TLR4-mediated disease or disorder, and wherein the siRNA targets a TLR4 signaling component.
6. The method of claim 1 , wherein the disease is a metabolic disease or disorder.
7. The method claim 1 or 6 , wherein the siRNA delivery system is administered systemically.
8. The method of claim 1 , wherein the disease or disorder is selected from the group consisting of an autoimmune disorder, sepsis, an intestinal disorder, an infectious disease, a malignancy, a pulmonary disorder, a cardiac disorder, and a neurological disorder.
9. The method of claim 8 , wherein the disease or disorder is selected from the group consisting of Crohn's disease, HIV, arthritis, cardiovascular disease and cancer.
10. The method of claim 1 or 6 , wherein the siRNA delivery system is administered orally.
11. The method of claim 6 , wherein the disease or disorder is selected from the group consisting of glucose tolerance, insulin sensitivity, diabetes and obesity.
12. The method of claim 6 , wherein the siRNA targets aP2.
13. The method of claim 6 , wherein the siRNA targets Map4k4, under conditions facilitating delivery of the siRNA to macrophage, wherein silencing of Map4k4 results in enhanced insulin sensitivity in the subject.
14. The method of claim 12 or 13 , wherein the siRNA delivery system is administered orally.
15. The method claim 12 or 13 , wherein the siRNA delivery system is administered systemically.
16. The method of claim 12 or 13 , wherein the subject has Type 2 Diabetes.
17. The method of claim 1 or 6 , wherein the YCWPs comprise at least a second siRNA coated with a second trapping layer.
18. The method of claim 1 , wherein the core comprises a cationic detergent.
19. The method of claim 1 , wherein the core comprises an anionic polymer coated with a cationic polymer, such that the core is cationic on the surface.
20. The method of claim 1 , wherein the siRNA is a Map4k4 siRNA.
21. The method of claim 1 , wherein the siRNA is an inflammatory cytokine siRNA.
22. The method of claim 1 , wherein the siRNA is a TNFα siRNA.
23. The method of claim 1 , wherein the siRNA is a RIP140 siRNA.
24. The method of claim 1 , wherein the siRNA is a TLR4 siRNA.
25. The method of claim 1 , wherein the siRNA is an aP2 siRNA.
26. The method of claim 1 , wherein the core further comprises the siRNA.
27. The method of claim 25 , wherein the core and siRNA layer comprise the same siRNA.
28. The method of claim 25 , wherein the core and siRNA layer comprise distinct siRNAs.
29. The method of claim 1 , wherein the siRNA is present at a ratio of about 1×10 −7 to 10×10- 7 pmol siRNA per YCWP.
30. The method of claim 1 , wherein the trapping layer comprises a cationic polymer, a cationic detergent or a mixture thereof.
31. The method of claim 1 , wherein the core comprises a cationic polymer, a cationic detergent or a mixture thereof.
32. The method of claim 30 or 31 , wherein the cationic polymer is selected from the group consisting of chitosan, poly-L-lysine and polyethylenimine (PEI).
33. The method of claim 19 , wherein the anionic polymer is an alginate or a xanthan.
34. The method of claim 19 , wherein the anionic polymer is a nucleic acid.
35. The method of claim 34 , wherein the nucleic acid is transfer RNA (tRNA).
36. The method of claim 1 , wherein the YCWP is chemically modified to comprise a cell or tissue specific targeting agent.
37. The method of claim 1 , wherein the siRNA delivery system delivers from 0.1 to 100 μg/kg to the subject as a unit dose.
38. The method of claim 1 , wherein the siRNA delivery system delivers from 1 to 10 μg/kg to the subject as a unit dose.