Compositions and methods for nucleic acid transfer
Disclosed are methods for the enhancement of nucleic acid delivery systems. The methods may employ treatment with a compound and/or an RNAi molecule in combination with a nucleic acid to improve nucleic acid uptake into a cell. In particular, the disclosed methods may be useful for improved gene therapy techniques.
1. A method for enhancing uptake of a nucleic acid nanoparticle into a eukaryotic cell for administration of a therapeutic gene, comprising
a. contacting said eukaryotic cell with said nucleic acid nanoparticle (NNP); and
b. contacting said eukaryotic cell with a nanoparticle uptake enhancing agent selected from one or both of an RNAi molecule and a compound;
wherein said NNP comprises a nucleic acid encoding said therapeutic gene or active portion thereof, and a cationic polymer conjugated to polyethylene glycol;
wherein said cationic polymer is selected from a poly-lysine, a polyethyleneimine, and combinations thereof;
wherein RNAi molecule inhibits expression of a gene encoding a protein selected from keratin 13, APC protein, protocadherin 17, and spectrin alpha; and
wherein said compound is selected from geldanamycin, alvespimycin, entasobulin, androstanolone, spermine, cortisone, quercetin, acetohexamide, resveratrol, doxorubicin, ruxolitinib, roscovitine, sildenafil, teniposide, etoposide, 3-Aminobenzamide (INO 1001), diazoxide, tedisamil, glimepiride, epirubicin, annamycin, repaglinide, 4β-aminoalkyl-4′-O-demethyl-4-desoxypodophyllotoxin (TOP53), acetohexamide, elsamitrucin, ketamine, NK109, tifenazoxide, olaparib, intoplicine, and combinations thereof, As 2 O 3 .
2. The method of claim 1 , wherein said active agent is selected from roscovitine, geldanamycin, acetohexamide, and ruxolitinib, or a combination thereof.
3. The method of claim 1 , wherein said NNP comprises a lysine polymer conjugated to PEG.
4. The method of claim 1 , wherein a period of time exists between step a and step b.
5. The method of claim 1 , wherein said nucleic acid is single stranded.
6. The method of claim 1 , wherein said nucleic acid is double stranded.
7. The method of claim 1 , wherein said method is carried out in vitro.
8. The method of claim 1 , wherein said method is carried out ex vivo.
9. A method of treating an individual, comprising the step of
a. administering an RNAi that inhibits expression of one or more genes encoding a protein selected from keratin 13, APC protein, protocadherin 17, spectrin alpha, and combinations thereof, and a compound selected from geldanamycin, alvespimycin, entasobulin, androstanolone, spermine, cortisone, quercetin, acetohexamide, resveratrol, doxorubicin, ruxolitinib, roscovitine, sildenafil, teniposide, etoposide, 3-Aminobenzamide (INO 1001), diazoxide, tedisamil, glimepiride, epirubicin, annamycin, repaglinide, 4β-aminoalkyl-4′-O-demethyl-4-desoxypodophyllotoxin (TOP53), acetohexamide, elsamitrucin, ketamine, NK109, tifenazoxide, olaparib, intoplicine, As 2 O 3 ; and
b. administering a nucleic acid nanoparticle (NNP) comprising a therapeutic gene, or portion of a therapeutic gene that encodes an active portion of a protein, that is expressed in said individual to provide a functional protein or functional protein fragment, wherein said functional protein or functional protein fragment is expressed in said individual in a therapeutically effective amount;
wherein said NNP comprises a cationic polymer conjugated to polyethylene glycol and;
wherein said cationic polymer is selected from a poly lysine, a polyethyleneimine, and combinations thereof.
10. The method of claim 9 , wherein said one or both of said RNAi and said compound are administered concurrently, before, or after administration of said NNP.
11. The method of claim 1 , wherein said NNP comprises a viral vector.
12. The method of claim 1 , wherein said NNP comprises a liposomal vector.
13. The method of claim 1 , wherein said NNP comprises a lipid complex.
14. The method of claim 1 , wherein said NNP comprises nucleic acid components of a CRISPR/CAS9 system.