Nanoparticle modification of human adipose-derived mesenchymal stem cells for treating brain cancer and other neurological diseases

Abstract

The presently disclosed subject matter provides compositions, methods, and kits for transfecting adipose-derived mesenchymal stem cells (AMSCs) in freshly extracted adipose tissue using nanoparticles comprising biodegradable polymers self-assembled with nucleic acid molecules. The presently disclosed subject matter also provides methods for treating a neurological disease in a patient in need thereof, the method comprising administering the AMSCs transfected with the nucleic acid molecules to the patient, wherein the nucleic acid molecules encode one or more bioactive molecules functional in the treatment of a neurological disease, particularly wherein the neurological disease is a brain tumor.

1 . A nanoparticle formulation comprising biodegradable polymers self-assembled with nucleic acid molecules, wherein the biodegradable polymer is selected from the group consisting of 2-(3-aminopropylamino)ethanol end-modified poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) (446), (1-(3-aminopropyl)-4-methylpiperazine end-modified poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) (447), 2-(3-aminopropylamino)ethanol end-modified poly(1,4-butanediol diacrylateco-5-amino-1-pentanol) (456), (1-(3-aminopropyl)-4-methylpiperazine end-modified poly(1,4-butanediol diacrylate-co-5-amino-1-pentanol) (457), 2-(3-aminopropylamino)ethanol end-modified poly(1,5 pentanediol diacrylate-co-3-amino-1-propanol) (536), and (1-(3-aminopropyl)-4-methylpiperazine-end-modified poly(1,5 pentanediol diacrylate-co-3-amino-1-propanol) (537), having a polymer to nucleic acid mass ratio of 25 to 75 weight polymer:weight nucleic acid, and wherein the nucleic acid molecules encode one or more bioactive molecules functional in the treatment of a neurological disease. 
     
     
         2 .- 5 . (canceled) 
     
     
         6 . The nanoparticle formulation of  claim 1 , wherein the one or more bioactive molecules are selected from the group consisting of proteins, polypeptides, peptides, drugs, enzymes, hormones, RNA, and metabolites. 
     
     
         7 . The nanoparticle formulation of  claim 6 , wherein the neurological disease is a brain tumor, and wherein the one or more bioactive molecules comprise one or more anti-cancer agents. 
     
     
         8 . The nanoparticle formulation of  claim 7 , wherein the one or more anti-cancer agents are selected from the group consisting of bone morphogenic protein 4 (BMP4), TNF-related apoptosis-inducing ligand (TRAIL), HSV-thymidine kinase, an oncolytic adenovirus, interleukin-2 (IL-2), interleukin-12 (IL-12), interleukin-18 (IL-18), interleukin-23 (IL-23), Interferon-α, and Interferon-β. 
     
     
         9 . The nanoparticle formulation of  claim 8 , wherein the nanoparticle formulation is lyophilized. 
     
     
         10 . A freshly extracted adipose tissue cell comprising at least one nanoparticle, wherein the nanoparticle comprises a biodegradable polymer self-assembled with a nucleic acid molecule, wherein the biodegradable polymer is selected from the group consisting of 2-(3-aminopropylamino)ethanol end-modified poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) (446), (1-(3-aminopropyl)-4-methylpiperazine end-modified poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) (447), 2-(3-aminopropylamino)ethanol end-modified poly(1,4-butanediol diacrylateco-5-amino-1-pentanol) (456), (1-(3-aminopropyl)-4-methylpiperazine end-modified poly(1,4-butanediol diacrylate-co-5-amino-1-pentanol) (457), 2-(3-aminopropylamino)ethanol end-modified poly(1,5 pentanediol diacrylate-co-3-amino-1-propanol) (536), and (1-(3-aminopropyl)-4-methylpiperazine-end-modified poly(1,5 pentanediol diacrylate-co-3-amino-1-propanol) (537), having a polymer to nucleic acid mass ratio of 25 to 75 weight polymer:weight nucleic acid, and wherein the nucleic acid molecules encode one or more bioactive molecules functional in the treatment of a neurological disease. 
     
     
         11 . The freshly extracted adipose tissue cell of  claim 10 , wherein the freshly extracted adipose tissue cell is an adipose-derived mesenchymal stem cell (AMSC). 
     
     
         12 .- 15 . (canceled) 
     
     
         16 . The freshly extracted adipose tissue cell of  claim 11 , wherein the one or more bioactive molecules are selected from the group consisting of proteins, polypeptides, peptides, drugs, enzymes, hormones, RNA, and metabolites. 
     
     
         17 . The freshly extracted adipose tissue cell of  claim 16 , wherein the neurological disease is a brain tumor, and wherein the one or more bioactive molecules comprise one or more anti-cancer agents. 
     
     
         18 . The freshly extracted adipose tissue cell of  claim 17 , wherein the one or more anti-cancer agents are selected from the group consisting of bone morphogenic protein 4 (BMP4), TNF-related apoptosis-inducing ligand (TRAIL), HSV-thymidine kinase, an oncolytic adenovirus, interleukin-2 (IL-2), interleukin-12 (IL-12), interleukin-18 (IL-18), interleukin-23 (IL-23), Interferon-α, and Interferon-β. 
     
     
         19 . A method for treating a neurological disease in a patient in need thereof, the method comprising:
 a. obtaining freshly extracted adipose tissue from the patient;   b. transfecting cells in the freshly extracted adipose tissue, wherein transfecting comprises combining the freshly extracted adipose tissue with a nanoparticle formulation to form a suspension, wherein the nanoparticle formulation comprises biodegradable polymers self-assembled with nucleic acid molecules, wherein the biodegradable polymer is selected from the group consisting of 2-(3-aminopropylamino)ethanol end-modified poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) (446), (1-(3-aminopropyl)-4-methylpiperazine end-modified poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) (447), 2-(3-aminopropylamino)ethanol end-modified poly(1,4-butanediol diacrylateco-5-amino-1-pentanol) (456), (1-(3-aminopropyl)-4-methylpiperazine end-modified poly(1,4-butanediol diacrylate-co-5-amino-1-pentanol) (457), 2-(3-aminopropylamino)ethanol end-modified poly(1,5 pentanediol diacrylate-co-3-amino-1-propanol) (536), and (1-(3-aminopropyl)-4-methylpiperazine-end-modified poly(1,5 pentanediol diacrylate-co-3-amino-1-propanol) (537), having a polymer to nucleic acid mass ratio of 25 to 75 weight polymer:weight nucleic acid, and wherein the nucleic acid molecules encode one or more bioactive molecules functional in the treatment of a neurological disease;   c. extracting a cellular fraction from the suspension, wherein the cellular fraction comprises freshly extracted adipose tissue cells transfected with the nucleic acid molecules; and   d. administering the freshly extracted adipose tissue cells transfected with the nucleic acid molecules to the patient.   
     
     
         20 . The method of  claim 19 , wherein the freshly extracted adipose tissue cells transfected with the nucleic acid molecules comprise adipose-derived mesenchymal stem cells (AMSCs). 
     
     
         21 .- 23 . (canceled) 
     
     
         24 . The method of  claim 20 , wherein the nucleic acid molecules encode one or more bioactive molecules functional in the treatment of a neurological disease. 
     
     
         25 . The method of  claim 24 , wherein the one or more bioactive molecules are selected from the group consisting of proteins, polypeptides, peptides, drugs, enzymes, hormones, RNA, and metabolites. 
     
     
         26 . The method of  claim 24 , wherein the neurological disease is a brain tumor, and wherein the one or more bioactive molecules comprise one or more anti-cancer agents. 
     
     
         27 . The method of  claim 26 , wherein the one or more anti-cancer agents are selected from the group consisting of bone morphogenic protein 4 (BMP4), TNF related apoptosis-inducing ligand (TRAIL), HSV-thymidine kinase, an oncolytic adenovirus, interleukin-2 (IL-2), interleukin-12 (IL-12), interleukin-18 (IL-18), interleukin-23 (IL-23), Interferon-α, and Interferon-β. 
     
     
         28 . The method of  claim 27 , wherein the freshly extracted adipose tissue cells transfected with the nucleic acid molecules are administered to the patient systemically. 
     
     
         29 . The method of  claim 28 , wherein the freshly extracted adipose tissue cells transfected with the nucleic acid molecules are administered to the patient intracranially. 
     
     
         30 . The method of  claim 29 , wherein the nanoparticle formulation is lyophilized prior to combining with the freshly extracted adipose tissue to form a suspension. 
     
     
         31 . A method for treating a neurological disease in a patient in need thereof, the method comprising:
 a. obtaining freshly extracted adipose tissue from the patient, wherein the freshly extracted adipose tissue comprises adipose-derived mesenchymal stem cells (AMSCs);   b. isolating and purifying the AMSCs;   c. culturing the AMSCs under conditions allowing for proliferation of the AMSCs;   d. transfecting the AMSCs, wherein transfecting comprises combining the freshly extracted adipose tissue with a nanoparticle formulation, wherein the nanoparticle formulation comprises biodegradable polymers self-assembled with nucleic acid molecules, wherein the biodegradable polymer is selected from the group consisting of 2-(3-aminopropylamino)ethanol end-modified poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) (446), (1-(3-aminopropyl)-4-methylpiperazine end-modified poly(1,4-butanediol diacrylate-co-4-amino-1-butanol) (447), 2-(3-aminopropylamino)ethanol end-modified poly(1,4-butanediol diacrylateco-5-amino-1-pentanol) (456), (1-(3-aminopropyl)-4-methylpiperazine end-modified poly(1,4-butanediol diacrylate-co-5-amino-1-pentanol) (457), 2-(3-aminopropylamino)ethanol end-modified poly(1,5 pentanediol diacrylate-co-3-amino-1-propanol) (536), and (1-(3-aminopropyl)-4-methylpiperazine-end-modified poly(1,5 pentanediol diacrylate-co-3-amino-1-propanol) (537), having a polymer to nucleic acid mass ratio of 25 to 75 weight polymer:weight nucleic acid, and wherein the nucleic acid molecules encode one or more bioactive molecules functional in the treatment of a neurological disease; and   e. administering the AMSCs transfected with the nucleic acid molecules to the patient.   
     
     
         32 .- 34 . (canceled) 
     
     
         35 . The method of  claim 31 , wherein the nucleic acid molecules encode one or more bioactive molecules functional in the treatment of a neurological disease. 
     
     
         36 . The method of  claim 35 , wherein the one or more bioactive molecules are selected from the group consisting of proteins, polypeptides, peptides, drugs, enzymes, hormones, RNA, and metabolites. 
     
     
         37 . The method of  claim 35 , wherein the neurological disease is a brain tumor, and wherein the one or more bioactive molecules comprise one or more anti-cancer agents. 
     
     
         38 . The method of  claim 37 , wherein the one or more anti-cancer agents are selected from the group consisting of bone morphogenic protein 4 (BMP4), TNF related apoptosis-inducing ligand (TRAIL), HSV-thymidine kinase, an oncolytic adenovirus, interleukin-2 (IL-2), interleukin-12 (IL-12), interleukin-18 (IL-18), interleukin-23 (IL-23), Interferon-α, and Interferon-β. 
     
     
         39 . The method of  claim 38 , wherein the AMSCs transfected with the nucleic acid molecules are administered to the patient systemically. 
     
     
         40 . The method of  claim 39 , wherein the AMSCs transfected with the nucleic acid molecules are administered to the patient in combination with an additional therapeutic agent or treatment. 
     
     
         41 . The method of  claim 40 , wherein the neurological disease is a brain tumor, and wherein the additional therapeutic agent or treatment is radiotherapy. 
     
     
         42 .- 50 . (canceled)