Methods and Compositions for Cellular Reprogramming

Abstract

Compositions and methods useful for the treatment of aberrant programming diseases, particularly those associated with aberrant apoptosis are disclosed

1 . A method for inhibiting apoptosis in a cell or tissue comprising:
 a) contacting said cells or tissue with an effective amount of at least one agent which inhibits p53 dependent cell death under conditions whereby said agent enters said cells and reduces apoptosis relative to untreated cells.   
     
     
         2 . The method of  claim 1 , wherein at least one said agent is a nucleic acid molecule, said nucleic acid molecule optionally comprising at least one modification which improves pharmacologic function. 
     
     
         3 . The method of  claim 1  or  2 , comprising co-administration of at least one augmentation agent selected from the group consisting of a free radical generator and a redox modifier. 
     
     
         4 . The method of  claim 1  or  2 , comprising co-administration of at least one augmentation agent selected from the group consisting of an antioxidant, anti-inflammatory agent and a cytokine. 
     
     
         5 . The method of  claim 1  or  2 , wherein said cells are normal cells adversely affected by anti-cancer treatment for an existing malignancy which treatment activates p53 dependent cell death. 
     
     
         6 . The method of  claim 5 , wherein said treatment is radiation or chemotherapy. 
     
     
         7 . The method of  claim 1  or  2  wherein said cells are damaged by a process selected from the group consisting of Alzheimer's disease, autoimmune disease, artherosclerosis, restenosis, ischemia/reperfusion injury, rheumatoid arthritis, arthritis, multiple sclerosis, lupus erythematosus, multiple organ dysfunction syndrome, myocardial infarction, stroke, ionizing radiation, and viral disease. 
     
     
         8 . The method of  claim 1  or  2 , wherein said cell or tissue is heart, skin, brain or a hematopoietic progenitor cell. 
     
     
         9 . The method of  claim 1  or  2 , wherein said at least one agent is a nucleic acid which is complementary to a portion of a p53 gene and is selected from the group consisting of an oligodeoxy-ribonucleotide, and an oligoribonucleotide. 
     
     
         10 . The method of  claim 9 , wherein said nucleic acid is a double stranded. 
     
     
         11 . The method of  claim 9 , wherein said nucleic acid is present in an expression vector. 
     
     
         12 . The method of  claim 9 , wherein uptake of said nucleic acid into said cell is facilitated by a carrier. 
     
     
         13 . The method of  claim 2 , wherein said nucleic acid binds to a transcriptional regulator binding site. 
     
     
         14 . The method of  claim 9 , wherein said nucleic acid is single stranded. 
     
     
         15 . The method of  claim 1  or  2 , wherein said method further comprises subjecting cells treated in a) to a Reprogramming test. 
     
     
         16 . The method of  claim 1  or  2 , wherein said nucleic acid is a double stranded oligoribonucleotide comprising a sequence complementary to a sequence encoding a transcriptional regulator. 
     
     
         17 . The method of  claim 9 , wherein said nucleic acid analog comprises a sequence selected from the group consisting of SEQ ID NOS: 1-4 and 863-872. 
     
     
         18 . The method of  claims 2  or  10 , wherein said modification is selected from the group consisting of a ethyl- or methylphosphonate modified oligodeoxynucleotides, phosphorothioate modified oligonucleotides, dithioates, oligonucleotide analogs, oligonucleotides comprising ribozymes, oligoribonucleotides, chimeric oligonucleotides that are composite RNA, DNA analogues, oligonucleotides having a lipophilic backbone, methylphosphonate analogs with ribozyme structures, and oligonucleotides with 2′-O-methyl modified ribose sugars. 
     
     
         19 . The method of  claims 1 ,  2  or  9 , said method further comprising administration of at least one agent or nucleic acid which inhibits expression of a target selected from the group consisting of apo-1 (SEQ ID NOS: 3374-3443), bax alpha (SEQ ID NO: 1027-1042), bcl-beta (SEQ ID NOS: 3610-3614), bcl-xl (SEQ ID NOS: 1000-1014), bcl-x (SEQ ID NOS: 1015-1026), cyclin A (SEQ ID NOS: 2076-2099), cyclin B (SEQ ID NO: 2066-2075), cyclin D exon 1 (SEQ ID NOS: 3589-3597), cyclin D exon 2 (SEQ ID NOS: 3598-3603), cyclin D exon 3 (SEQ ID NO: 3604-3605, cyclin D exon 4 (SEQ ID NOS: 3606-3609), cyclin D3 exon 1 (SEQ ID NO: 3587-3588), cyclin D3 exon 3 (SEQ ID NOS: 3580-3586), cyclin D3 exon 4 (SEQ ID NOS: 3575-3579) cyclin D1 (SEQ ID NOS: 727-738), CREBP-1 (SEQ ID NOS: 326-353), ICE (SEQ ID NOS: 2788-2802), ICH-1L (SEQ ID NOS: 1326-1343), TGF-beta (SEQ ID NOS: 3717-3741, TNF-alpha (SEQ ID NOS: 3145-3176), TNF-beta (SEQ ID NOS: 3112-3144, TR3 (SEQ ID NOS: 3216-3241), E2F-1 (SEQ ID NOS: 221-234), GADD153 (SEQ ID NOS: 1344-1353), IRF-1 (SEQ ID NOS: 271-289), ISGF-3 (SEQ ID NOS: 1051-1068), MSX2 (SEQ ID NO: 1481-1491), NF-IL-6 (SEQ ID NOS: 2803-2852), GADD45 (SEQ ID NOS: 1354-1404), c-jun (SEQ ID NOS:891-908), junB (SEQ ID NOS: 455-464), junD (SEQ ID NOS: 5-17, 396, 399, 401, 403, 405, 407, 409, 412, 414, 416, and 418), c-fos (SEQ ID NOS: 465-482), Fra-1 (SEQ ID NOS: 449-454), Fra-2 (SEQ ID NOS: 627-638), PKC-alpha (SEQ ID NOS: 1869-1881), PKC-beta (SEQ ID NOS: 1882-1887), PKC-delta (SEQ ID NO: 1898-1917), CREBP-1 (SEQ ID NOS: 326-353), PKC-epsilon (SEQ ID NOS: 1918-1937), PKC-gamma (SEQ ID NOS: 1888-1897) PKC-iota (SEQ ID NOS: 1462-1480), PKC-mu (SEQ ID NOS: 1442-1461), PKC-theta (SEQ ID NOS: 3615-3626), PKC-zeta (SEQ ID NOS: 1417-1441) and c-myc (SEQ ID NOS: 657-676). 
     
     
         20 . A pharmaceutical composition comprising the agent of  claims 1 ,  2  or  9  in a pharmaceutically acceptable carrier. 
     
     
         21 . The pharmaceutical composition of  claim 20 , further comprising an effective amount of at least one of a free radical generator and a redox modifier. 
     
     
         22 . The pharmaceutical composition of  claim 20  further comprising at least one of an antioxidant, an anti-inflammatory and a cytokine. 
     
     
         23 . The pharmaceutical composition of  claim 20 , wherein said nucleic acid is selected from the group consisting of SEQ ID NOS: 1-4 and 863-872. 
     
     
         24 . The method of  claims 1  or  2 , wherein said cells are damaged as a result of blood vessel occlusion followed by reperfusion injury. 
     
     
         25 . The method of  claims 1  or  2 , or  9  or  19  wherein said method is for treatment of apoptosis associated with viral infection, wherein said virus is selected from the group consisting of adenovirus, cytomegalovirus, Epstein-Barr virus, hepatitis C virus, herpes virus, hemorrhagic fever viruses, human immunodeficiency virus, influenza virus, pox virus, vaccinia virus. 
     
     
         26 . The method of  claim 25 , further comprising administration of at least one augmentation agent selected from the group consisting of an anti-oxidant, a redox modifier, a cytokine and an interferon. 
     
     
         27 . The method of  claim 25 , wherein said virus is HIV and said nucleic acid inhibits virus expression or effects of virus on cells and is selected from the group consisting of GADD-153 (SEQ ID NOS: 1344-1353), GADD-45 (SEQ ID NO: 1354-1404), MTS 1,2, (SEQ ID NOS: 2454-2472 and 2100-28), TGF beta (SEQ ID NOS: 3717-3741), USF (SEQ ID NOS:1826-1852), Ap-2 (SEQ ID NOS: 1252-1289), Ap-4 (SEQ ID NOS: 2242-2264), Sp-1 (SEQ ID NOS: 989-999), Sp-3 (SEQ ID NOS: 985-988), Sp-4 (SEQ ID NOS: 978-984), p53 (SEQ ID NOS:863-872), NF-kappa B (SEQ ID NOS:1741-1774, 1720-1739, 2166-2205, 2007-2011), rel (SEQ ID NOS:437-448), GATA-3 (SEQ ID NOS: 2992-3008), and Waf1 (SEQ ID NOS: 2440-2453). 
     
     
         28 . The method of  claim 27 , wherein said viral infection causes AIDS.