Lipoproteins as nucleic acid vectors

Abstract

The present invention relates to materials and methods for the in vivo transport and deliver of nucleic acids. More particularly, it concerns the use of lipoproteins, including but not limited to, low density lipoproteins (“LDL”), and/or apolipoproteins for the binding and in vivo transport of nucleic acids. In addition, the present invention relates to the use of lipoproteins in the early detection of cancer and/or metastatic cancer and/or arteriosclerosis.

1 . A composition comprising: 
 (a) an isolated polypeptide comprising at least one LDL or VLDL nucleic acid binding domain; and    (b) a nucleic acid comprising an LDL or VLDL binding sequence,    wherein said nucleic acid is bound to said polypeptide.    
     
     
         2 . The composition of  claim 1 , wherein said polypeptide comprises an LDL nucleic acid binding domain.  
     
     
         3 . The composition of  claim 1 , wherein said polypeptide comprises a VLDL nucleic acid binding domain.  
     
     
         4 . The composition of  claim 1 , wherein said nucleic acid comprises an expression region operably linked to a promoter active in eukaryotic cells.  
     
     
         5 . The composition of  claim 4 , wherein said expression region encodes a polypeptide.  
     
     
         6 . The composition of  claim 4 , wherein said expression region comprises an antisense construct.  
     
     
         7 . The composition of  claim 5 , wherein said polypeptide is selected from the group consisting of α-globin, β-globin, γ-globin, granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor (TNF), IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, β-interferon, γ-interferon, cytosine deaminase, adenosine deaminase, β-glucuronidase, hypoxanthine guanine phosphoribosyl transferase, galactose-1-phosphate uridyltransferase, glucocerbrosidase, glucose-6-phosphatase, thymidine kinase, lysosomal glucosidase, growth hormone, nerve growth factor, insulin, adrenocorticotropic hormone, parathormone, follicle-stimulating hormone, luteinizing hormone, epidermal growth factor, thyroid stimulating hormone, CFTR, EGFR, VEGFR, IL-2 receptor, estrogen receptor, Bax, Bak, Bcl-X s , Bik, Bid, Bad, Harakiri, Ad E1B, an ICE-CED3 protease neomycin resistance, luciferase, adenine phosphoribosyl transferase (APRT), retinoblastoma, insulin, mast cell growth factor, p53, p16, p21, MMAC1, p73, zacl and BRCAI.  
     
     
         8 . The composition of  claim 6 , wherein said antisense construct is complementary to a segment of an oncogene.  
     
     
         9 . The composition of  claim 8 , wherein said oncogene is selected from the group consisting of ras, myc, neu, raf, erb, src, fms, jun, trk, ret, gsp, hst, bcl and abl.  
     
     
         10 . The composition of  claim 4 , wherein said promoter is selected from the group consisting of CMV IE, LTR, SV40 IE, HSV trk, β-actin, human globin α, human globin β and human globin γ promoter.  
     
     
         11 . The composition of  claim 1 , wherein said nucleic acid binding domain is an apoB100 nucleic acid binding domain.  
     
     
         12 . The composition of  claim 1 , wherein said composition further comprises one or more lipoproteins selected from the group consisting of apoA1, apoA-II, apoA-IV, acat, apoE, apoC-II, apoC-III and apo-D.  
     
     
         13 . The composition of  claim 11 , wherein said apoB100 is selected from the group consisting of human, rat and baboon apoB100.  
     
     
         14 . The composition of  claim 1 , wherein said polypeptide comprises at least two nucleic acid binding domains.  
     
     
         15 . The composition of  claim 14 , wherein said nucleic acid binding domain contains a motif selected from the group consisting of a proline pipe helix DNA binding motif, a ISGF3γ-like DNA binding motif, a SREBP-like DNA binding motif, a coiled-coil motif and a nucleotide (ATP)-binding motif.  
     
     
         16 . The composition of  claim 14 , wherein said binding domain is selected from the group consisting of SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100,, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:105, SEQ ID NO:106, SEQ ID NO:107, SEQ ID NO:108, SEQ ID NO:109, SEQ ID NO:110, SEQ ID NO:111, SEQ ID NO:112, NO:113, SEQ ID NO:114, SEQ ID NO:115, SEQ ID NO:144, SEQ ID NO:145, SEQ ID NO:146, SEQ ID NO:147, SEQ ID NO:148, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:151, SEQ ID NO:152, SEQ ID NO:153, SEQ ID NO:154, SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:165, SEQ ID NO:166 and SEQ ID NO:175.  
     
     
         17 . The composition of  claim 1 , wherein said polypeptide further comprises at least one nuclear localization sequence.  
     
     
         18 . The composition of  claim 17 , wherein said nuclear localization sequence is from apoB100.  
     
     
         19 . The composition of  claim 17 , wherein said nuclear localization sequence is selected from the group consisting of SEQ ID NO:178, SEQ ID NO: 179, SEQ ID NO: 180, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 208, SEQ ID NO: 209, SEQ ID NO: 210.  
     
     
         20 - 63 . (canceled)  
     
     
         64 . A pharmaceutical composition comprising: 
 (a) an isolated polypeptide comprising at least one LDL or VLDL nucleic acid binding domain; and    (b) a nucleic acid comprising an LDL or VLDL binding sequence, wherein said nucleic acid is bound to said polypeptide;    said pharmaceutical composition being dispersed in a suitable diluent.    
     
     
         65 . A method of transforming a cell comprising: 
 a) providing a cell;    b) contacting said cell with a composition comprising (i) an isolated polypeptide comprising at least one LDL or VLDL nucleic acid binding domain and (ii) an expression cassette comprising a nucleic acid sequence encoding an expression region and a promoter active in eukaryotic cells, wherein said expression region is operably linked to said promoter, and wherein said nucleic acid sequence is bound to said LDL or VLDL;    wherein expression of said expression region is indicative of said transformation.    
     
     
         66 . A method of transfecting a cell comprising the steps of: 
 a) providing a cell;    b) contacting said cell with a composition comprising (i) an isolated polypeptide comprising at least one LDL or VLDL nucleic acid binding domain and (ii) an