Lipid nanoparticles and use thereof to deliver RNA polynucleotides

1. A lipid nanoparticle comprising 1,2-dioleoyl-3-trimethylammonium propane (DOTAP), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol, and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000), wherein the molar ratio of DOTAP: DSPC: cholesterol: DSPE-PEG2000 is 40:10:48:2.

2. The lipid nanoparticle of claim 1 , consisting of DOTAP: DSPC:

cholesterol:DSPE-PEG2000 at a molar ratio of 40:10:48:2.

3. The lipid nanoparticle of claim 1 , wherein the lipid nanoparticle is about 100 nM in diameter.

4. The lipid nanoparticle of claim 1 , wherein the surface charge of the lipid nanoparticle is about neutral to about 25 mV.

5. A composition comprising the lipid nanoparticle of claim 1 and one or more RNA polynucleotide.

6. The composition of claim 5 , wherein the one or more RNA polynucleotide comprises one or more self-replicating RNA derived from an alphavirus.

7. The composition of claim 6 , wherein the alphavirus is a Venezuelan equine encephalitis virus, optionally wherein the Venezuelan equine encephalitis virus is TC-83 or a variant thereof.

8. The composition of claim 6 , wherein at least one of the self-replicating RNA comprises a 5′ cap.

9. The composition of claim 6 , wherein the ratio of cationic amines in the lipid nanoparticle to anionic phosphates in the self-replicating RNA (N:P ratio) is about 0.5:1 to about 20:1.

10. The composition of claim 6 , wherein at least one of the self-replicating RNA comprises a sequence encoding a therapeutic agent.

11. The composition of claim 10 , wherein the therapeutic agent is (i) a cytokine, a chemokine, or a growth factor, or (ii) an anti-tumor agent, optionally wherein the anti-tumor agent is interleukin-15 (IL-15) or an IL-15 superagonist.

12. The composition of claim 6 , wherein the composition comprises two or more self-replicating RNAs.

13. The composition of claim 6 , wherein the self-replicating RNA is present within the lipid nanoparticle.

14. A method of treating a disease or disorder in a subject in need thereof, comprising providing to the subject the composition of claim 10 , wherein the self-replicating RNA comprises the sequence encoding the therapeutic agent.

15. The method of claim 14 , wherein the disease or disorder is a cancer, and the therapeutic agent is an anti-tumor agent, optionally wherein the anti-tumor agent is interleukin-15 (IL-15) or an IL-15 superagonist.

16. The method of claim 15 , wherein the cancer is a a breast cancer, a head and neck cancer, a lymphoma or a skin cancer, optionally wherein the skin cancer is a melanoma.

17. The method of claim 14 , wherein the composition is provided to the subject systemically locally, or parenterally.

18. The method of claim 14 , wherein the composition is provided in one or more doses, optionally wherein the one or more doses are two or more doses provided within two weeks of each other.

19. The method of claim 14 , wherein the composition is provided at about 1 μg/dose to about 1000 μg/dose.

20. The method of claim 15 , wherein the treatment inhibits tumor growth by at least 10% as compared to growth of an untreated tumor.

21. The method of claim 17 , wherein the composition is provided to the subject intratumorally or intravenously.