Delivery of RNA to different cell types

Patent metadata

Field	Value
Publication number	US-9770463-B2
Application number	US-201113808087-A
Country	US
Kind code	B2
Filing date	Jun 7, 2011
Priority date	Jul 6, 2010
Publication date	Sep 26, 2017
Grant date	Sep 26, 2017

Abstract

RNA encoding an immunogen is co-delivered to non-immune cells at the site of delivery and also to immune cells which infiltrate the site of delivery. The responses of these two cell types to the same delivered RNA lead to two different effects, which interact to produce a strong immune response against the immunogen. The non-immune cells translate the RNA and express the immunogen. Infiltrating immune cells respond to the RNA by expressing type I interferons and pro-inflammatory cytokines which produce a local adjuvant effect which acts on the immunogen-expressing non-immune cells to upregulate major histocompatibility complex expression, thereby increasing presentation of the translated protein to T cells. The effects on the immune and non-immune cells can be achieved by a single delivery of a single RNA e.g. by a single injection.

First claim

The invention claimed is:

1. A method of raising an immune response in a vertebrate, comprising administering an immunogen-encoding self-replicating RNA to a delivery site in a vertebrate and then permitting immune cells to infiltrate the delivery site, such that the self-replicating RNA separately enters both (a) non-immune cells at the delivery site and (b) the infiltrating immune cells, provided that the self-replicating RNA does not comprise modified nucleotides other than a 5′ cap, and wherein said immunogen is translated by a host cell from said vertebrate, wherein the self-replicating RNA is administered without protein and in combination with a cationic lipid-containing delivery system comprising:
(i) liposomes comprising DSPC (1,2-Diastearoyl-sn-glycero-3-phosphocholine), DlinDMA (1,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane), cholesterol, and PEG-conjugated DMG (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)), or
(ii) a submicron cationic oil-in-water emulsion comprising squalene, span 85 (sorbitan trioleate), polysorbate 80, and DOTAP (1,2-dioleoyl-3-trimethylammonium-propane).

2. The method of claim 1 , wherein the delivery site is in skeletal muscle tissue.

3. The method of claim 1 , wherein the self-replicating RNA is administered by injection.

4. The method of claim 3 , wherein injection is via a needle.

5. The method of claim 1 , wherein the non-immune cells comprise muscle cells and/or fibroblasts.

6. The method of claim 1 , wherein the self-replicating RNA is translated by the non-immune cells.

7. The method of claim 1 , wherein the immune cells comprise macrophages, dendritic cells and/or monocytes.

8. The method of claim 1 , wherein the immune cells are TLR7-positive.

9. The method of claim 1 , wherein the self-replicating RNA causes the immune cells to secrete type I interferons and/or pro-inflammatory cytokines.

10. The method of claim 1 , wherein the self-replicating RNA is +-stranded and can be translated by the non-immune cells.

11. The method of claim 1 , wherein the self-replicating RNA can bind to TLR7.

12. The method of claim 1 , wherein the self-replicating RNA encodes an immunogen which can elicit an immune response against a bacterium, a virus, a fungus or a parasite.

13. The method of claim 12 , wherein the immunogen can elicit an immune response in vivo against respiratory syncytial virus glycoprotein F.

14. The method of claim 12 , wherein the immunogen can elicit an immune response in vivo against: (a) a virus which infects fish, such as infectious salmon anemia virus (ISAV), salmon pancreatic disease virus (SPDV), infectious pancreatic necrosis virus (IPNV), channel catfish virus (CCV), fish lymphocystis disease virus (FLDV), infectious hematopoietic necrosis virus (IHNV), koi herpesvirus, salmon picorna-like virus (also known as picorna-like virus of atlantic salmon), landlocked salmon virus (LSV), atlantic salmon rotavirus (ASR), trout strawberry disease virus (TSD), coho salmon tumor virus (CSTV), and viral hemorrhagic septicemia virus (VHSV); (b) orthomyxovirus, such as influenza A, influenza B and influenza C virus; or (c) herpesvirus, such as herpes simplex viruses (HSV), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), human herpesvirus 6 (HHV6), human herpesvirus 7 (HHV7), and human herpesvirus 8 (HHV8).

15. The method of claim 12 , wherein the immunogen can elicit an immune response in vivo against Neisseria meningitidis.

16. The method of claim 1 , wherein the self-replicating RNA encodes (i) an alphavirus replicase comprising one or more of alphavirus proteins nsP1, nsP2, nsP3, and nsP4 that can transcribe RNA from the self-replicating RNA molecule and (ii) the immunogen.

17. A method of delivering an immunogen-encoding self-replicating RNA to both (a) nonimmune cells at a delivery site and (b) infiltrating immune cells in a vertebrate, comprising administering said self-replicating RNA by intramuscular injection to the delivery site in the vertebrate and then permitting immune cells to infiltrate the delivery site, provided that the self-replicating RNA includes no modified nucleotides other than a 5′ cap, is not packaged as a virion, and cannot induce production of RNA-containing virions, wherein the RNA is administered without protein and in combination with a delivery system comprising:
(i) liposomes comprising DSPC (1,2-Diastearoyl-sn-glycero-3-phosphocholine), DlinDMA (1,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane), cholesterol, and PEG-conjugated DMG (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N4methoxy(polyethylene glycol)), or
(ii) a submicron cationic oil-in-water emulsion comprising squalene, span 85 (sorbitan trioleate), polysorbate 80, and DOTAP (1,2-dioleoyl-3-trimethylammonium-propane).

18. A method of activating a cytoplasmic RNA helicase and stimulating a type I interferon and/or a pro-inflammatory cytokine in a vertebrate, comprising administering an immunogen-encoding self-replicating RNA to a delivery site in the vertebrate and then permitting immune cells to infiltrate the delivery site, provided that the self-replicating RNA includes no modified nucleotides other than a 5′ cap, is not packaged as a virion, and cannot induce production of RNA-containing virions, wherein the RNA is administered without protein and in combination with a delivery system comprising:
(i) liposomes comprising DSPC (1,2-Diastearoyl-sn-glycero-3-phosphocholine), DlinDMA (1,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane), cholesterol, and PEG-conjugated DMG (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)), or
(ii) a submicron cationic oil-in-water emulsion comprising squalene, span 85 (sorbitan trioleate), polysorbate 80, and DOTAP (1,2-dioleoyl-3-trimethylammonium-propane).

Assignees

• Geall Andrew
• Ramsauer Katrin
• Otten Gillis
• Mandl Christian
• Glaxosmithkline Biologicals Sa

Inventors

• Geall Andrew
• Ramsauer Katrin
• Otten Gillis
• Mandl Christian

Classifications

• C12N2770/36143

  viral genome or elements thereof as genetic vector · CPC title

• A61K2039/55505

  Inorganic adjuvants · CPC title

• C12N2760/18534

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• A61K39/155

  Paramyxoviridae, e.g. parainfluenza virus · CPC title

• A61K31/7088 Primary

  Compounds having three or more nucleosides or nucleotides · CPC title