Modified adipose tissue and related implants and methods

Abstract

The present invention provides adipose tissue modified with a vector comprising or encoding, in which case it expresses, an anti-angiogenic factor, an angiogenic substance, an apoptotic factor, an adipsin protein or an Ob protein. Also provided is adipose tissue modified with a vector comprising a promoter and, operably linked thereto, a DNA sequence encoding a secreted protein. Methods of therapeutically treating adipose tissue and expressing a secreted protein in adipose tissue are also provided. The adipose tissue is optionally in the form of an implant, which can further comprise a lymphogenic protein or a vector that comprises and expresses a lymphogenic gene.

What is claimed is:  
     
         1 . A method of therapeutically treating adipose tissue, which method comprises contacting said adipose tissue with a vector comprising a nucleic acid sequence comprising or encoding, in which case it expresses, 
 a) an anti-angiogenic factor such that said vector enters said adipose tissue and said anti-angiogenic factor inhibits vascularity in said adipose tissue,    b) an apoptotic factor such that said vector enters said adipose tissue and said apoptotic factor causes adipocyte cell death,    c) an adipsin protein such that said vector enters said adipose tissue and said adipsin protein treats said adipose tissue therapeutically,    d) an Ob protein such that said vector enters said adipose tissue and said Ob protein treats said adipose tissue therapeutically, or    e) an angiogenic substance such that said vector enters said adipose tissue and said angiogenic substance increases the vascularity in said adipose tissue,    with the proviso that, when said vector comprises a nucleic acid sequence encoding any one of (a)-(e), said nucleic acid sequence is operably linked to a promoter.    
     
     
         2 . The method of  claim 1 , wherein said vector is a viral vector.  
     
     
         3 . The method of  claim 2 , wherein said viral vector is an adenoviral vector.  
     
     
         4 . The method of  claim 3 , wherein said adenoviral vector is replication-deficient.  
     
     
         5 . The method of  claim 1 , wherein said promoter is adipocyte-specific.  
     
     
         6 . The method of  claim 5 , wherein said promoter is from the regulatory region of either of the adipocyte P2 (aP2) gene or the p154 polypeptide gene.  
     
     
         7 . The method of  claim 1 , wherein said promoter is constitutive.  
     
     
         8 . The method of  claim 1 , wherein said anti-angiogenic factor is selected from the group consisting of taxol, endostatin, angiostatin, fumagillin and an analogue of fumagillin.  
     
     
         9 . The method of  claim 1 , wherein said angiogenic substance is a vascular endothelial growth factor (VEGF).  
     
     
         10 . The method of  claim 1 , wherein said apoptotic factor is selected from the group consisting of p53, a cell death-inducing coding sequence of Bc1-2 which comprises an N-terminal deletion, a cell death-inducing coding sequence of Bc1-x which comprises an N-terminal deletion, Bax, Bak, Bid, Bad, Bik, Bif-2, IAP-1, IAP 2, a caspase, TGF β1, c myc, a protease, and a protein kinase.  
     
     
         11 . The method of  claim 10 , wherein said protein kinase is selected from the group consisting of protein kinase Cθ, protein kinase Cδ, Akt/PI(3)-kinase, DNA-PK, PITSLRE, DAP kinase, RIP, JNK/SAPK, Daxx, Raf-1, Pim-1, NIK, MEKK1, ASK1, and PKR.  
     
     
         12 . An isolated adipose tissue comprising a vector comprising a nucleic acid sequence comprising or encoding, in which case it expresses, 
 a) an anti-angiogenic factor,    b) an apoptotic factor,    c) an adipsin protein,    d) an Ob protein, or    e) an angiogenic substance,    wherein said isolated adipose tissue is optionally in the form of an implant.    
     
     
         13 . The isolated adipose tissue of  claim 12 , wherein said vector is a viral vector.  
     
     
         14 . The isolated adipose tissue of  claim 13 , wherein said viral vector is an adenoviral vector.  
     
     
         15 . The isolated adipose tissue of  claim 14 , wherein said adenoviral vector is replication-deficient.  
     
     
         16 . The isolated adipose tissue of  claim 12 , wherein said promoter is adipocyte-specific.  
     
     
         17 . The isolated adipose tissue of  claim 12 , wherein said promoter is from the regulatory region of either of the adipocyte P2 (aP2) gene or the p154 polypeptide gene.  
     
     
         18 . The isolated adipose tissue of  claim 12 , wherein said promoter is constitutive.  
     
     
         19 . The isolated adipose tissue of  claim 12 , wherein said anti-angiogenic factor is selected from the group consisting of taxol, endostatin, angiostatin, fumagillin and an analogue of fumagillin.  
     
     
         20 . The isolated adipose tissue of  claim 12 , wherein said angiogenic substance, anti-angiogenic factor, adipsin protein or Ob protein is secreted.  
     
     
         21 . The isolated adipose tissue of  claim 12 , wherein said apoptotic gene is selected from the group consisting of p53, a cell death-inducing coding sequence of Bc1-2 which comprises an N-terminal deletion, a cell death-inducing coding sequence of Bc1-x which comprises an N-terminal deletion, Bax, Bak, Bid, Bad, Bik, Bif-2, IAP-1, IAP-2, a caspase, TGF-β1, c-myc, a protease, and a protein kinase.  
     
     
         22 . The isolated adipose tissue of  claim 21 , wherein said protein kinase is selected from the group consisting of protein kinase Cθ, protein kinase Cδ, Akt/PI(3)-kinase, DNA-PK, PITSLRE, DAP kinase, RIP, JNK/SAPK, Daxx, Raf-1, Pim-1, NIK, MEKK1, ASK1, and PKR.  
     
     
         23 . The isolated adipose tissue of  claim 12 , wherein said isolated adipose tissue is in the form of an implant and further comprises a lymphogenic protein or a vector that comprises and expresses a lymphogenic gene.  
     
     
         24 . The adipose tissue implant of  claim 12 , wherein said vector comprises a gene encoding a vascular endothelial growth factor (VEGF).  
     
     
         25 . A method of expressing a secreted protein in adipose tissue, which method comprises contacting said adipose tissue with a vector comprising a promoter and, operably linked thereto, a DNA sequence encoding a secreted protein such that said gene transfer vector enters said adipose tissue and said protein is expressed and secreted.  
     
     
         26 . The method of  claim 25 , wherein said vector is a viral vector.  
     
     
         27 . The method of  claim 26 , wherein said viral vector is an adenoviral vector.  
     
     
         28 . The method of  claim 27 , wherein said adenoviral vector is replication-deficient.  
     
     
         29 . The method of  claim 25 , wherein said promoter is adipocyte-specific.  
     
     
         30 . The method of  claim 29 , wherein said promoter is from the regulatory region of either of the adipocyte P2 (aP2) gene or the p154 polypeptide gene.  
     
     
         31 . The method of  claim 25 , wherein said promoter is constitutive.  
     
     
         32 . An isolated adipose tissue comprising a vector comprising a promoter and, operably linked thereto, a DNA sequence encoding a secreted protein, wherein said isolated adipose tissue is optionally in the form of an implant.  
     
     
         33 . The isolated adipose tissue of  claim 32 , wherein said vector is a viral vector.  
     
     
         34 . The isolated adipose tissue of  claim 33 , wherein said viral vector is an adenoviral vector.  
     
     
         35 . The isolated adipose tissue of  claim 34 , wherein said adenoviral vector is replication-deficient.  
     
     
         36 . The isolated adipose tissue of  claim 32 , wherein said promoter is adipocyte-specific.  
     
     
         37 . The isolated adipose tissue of  claim 32 , wherein said promoter is from the regulatory region of either of the adipocyte P2 (aP2) gene or the p154 polypeptide gene.  
     
     
         38 . The isolated adipose tissue of  claim 32 , wherein said promoter is constitutive.  
     
     
         39 . The isolated adipose tissue of  claim 32 , wherein said isolated adipose tissue is in the form of an implant and further comprises a lymphogenic protein or a vector that comprises and expresses a lymphogenic gene.