Method for selective expression of therapeutic genes by hyperthermia

Abstract

A method for controlling in vivo heat-inducible gene expression for provision of therapeutic polypeptide. The method includes the steps of: (a) providing a construct comprising a polynucleotide encoding a therapeutic polypeptide, wherein the polynucleotide is operatively linked to a heat-inducible promoter; (b) administering the construct to a warm-blooded vertebrate at a site at or near the tumor; and (c) applying heat to the site at or near the tumor to express the therapeutic polypeptide, whereby anti-tumor activity is observed.

1 . A method for promoting anti-tumor activity in a warm-blooded vertebrate, the method comprising: 
 (a) providing a construct comprising a polynucleotide encoding a therapeutic polypeptide, wherein the polynucleotide is operatively linked to a heat-inducible promoter;    (b) administering the construct to a warm-blooded vertebrate at a site at or near the tumor; and    (c) applying heat to the site at or near the tumor to express the therapeutic polypeptide, whereby anti-tumor activity is observed.    
     
     
         2 . The method of  claim 1 , wherein the polynucleotide encodes a polypeptide having an ability to elicit an immunostimulatory response, an anti-angiogenic response, an anti-proliferative response, a cell suicide response, or combinations thereof.  
     
     
         3 . The method of  claim 2  wherein the polynucleotide encodes a polypeptide having an ability to elicit both an immunostimulatory response and an anti-angiogenic response.  
     
     
         4 . The method of  claim 1 , wherein the polynucleotide encodes a functional IL2 polypeptide.  
     
     
         5 . The method of  claim 4 , wherein the IL2 polypeptide comprises: 
 (a) a polypeptide encoded by a nucleic acid molecule of SEQ ID NO:9 or 11;    (b) a polypeptide encoded by a nucleic acid substantially identical to SEQ ID NO:9 or 11;    (c) a polypeptide having an amino acid sequence set forth as SEQ ID NO:10 or 12;    (d) a polypeptide having an amino acid sequence substantially identical to SEQ ID NO:10 or 12; or    (e) a polypeptide that is a biological equivalent of SEQ ID NO:10 or 12.    
     
     
         6 . The method of  claim 4 , wherein the polynucleotide comprises: 
 (a) a nucleotide sequence set forth as SEQ ID NO:9 or 11; or    (b) a nucleotide sequence that is substantially identical to SEQ ID NO:9 or 11.    
     
     
         7 . The method of  claim 1 , wherein the heat-inducible promoter comprises an hsp70 promoter.  
     
     
         8 . The method of  claim 7 , wherein the hsp70 promoter comprises a human hsp70B promoter.  
     
     
         9 . The method of  claim 8 , wherein the hsp70B promoter comprises: 
 (a) the nucleotide sequence set forth as SEQ ID NO:13, or functional portion thereof; or    (b) a nucleotide sequence substantially identical to SEQ ID NO:13.    
     
     
         10 . The method of  claim 1 , wherein the anti-tumor activity is characterized by an immune response and an anti-angiogenic response.  
     
     
         11 . The method of  claim 1 , further comprising providing radiotherapy at the site at or near the tumor, whereby anti-tumor activity is enhanced when compared to anti-tumor activity observed in response to radiotherapy alone.  
     
     
         12 . A method for treating a disorder or condition in a warm-blooded vertebrate, the method comprising: 
 (a) providing a construct comprising a polynucleotide encoding a functional IL2 polypeptide, wherein the polynucleotide is operatively linked to a heat-inducible promoter;    (b) administering the construct to a warm-blooded vertebrate at or near a site in need of treatment; and    (c) applying heat to the site to express the polypeptide, whereby the disorder or condition is treated.    
     
     
         13 . The method of  claim 12 , wherein the disorder or condition is a proliferative or angiogenic disorder.  
     
     
         14 . The method of  claim 12 , wherein the IL2 polypeptide comprises: 
 (a) a polypeptide encoded by a nucleic acid molecule of SEQ ID NO:9 or 11;    (b) a polypeptide encoded by a nucleic acid substantially identical to SEQ ID NO:9 or 11;    (c) a polypeptide having an amino acid sequence set forth as SEQ ID NO:10 or 12;    (d) a polypeptide having an amino acid sequence substantially identical to SEQ ID NO:10 or 12; or    (e) a polypeptide that is a biological equivalent of SEQ ID NO:10 or 12.    
     
     
         15 . The method of  claim 12 , wherein the polynucleotide comprises: 
 (a) a nucleotide sequence set forth as SEQ ID NO:9 or 11; or    (b) a nucleotide sequence that is substantially identical to SEQ ID NO:9 or 11.    
     
     
         16 . The method of  claim 12 , wherein the heat-inducible promoter comprises an hsp70 promoter.  
     
     
         17 . The method of  claim 16 , wherein the hsp70 promoter comprises a human hsp70B promoter.  
     
     
         18 . The method of  claim 17 , wherein the hsp70B promoter comprises: 
 (a) the nucleotide sequence set forth as SEQ ID NO:13, or functional portion thereof; or    (b) a nucleotide sequence substantially identical to SEQ ID NO:13.    
     
     
         19 . The method of  claim 12 , wherein the site in need of treatment is a tumor.  
     
     
         20 . The method of  claim 19 , wherein treating the disorder comprises promoting tumor regression.  
     
     
         21 . The method of  claim 20 , further comprising providing radiotherapy at a site at or near a tumor, whereby anti-tumor activity is enhanced compared to anti-tumor activity in response to radiotherapy alone.  
     
     
         22 . A method for treating a disorder or condition in a warm-blooded vertebrate, the method comprising: 
 (a) providing a construct comprising a polynucleotide encoding a functional IL12 polypeptide, wherein the polynucleotide is operatively linked to a heat-inducible promoter;    (b) administering the construct to a warm-blooded vertebrate at or near a site in need of treatment; and    (c) applying heat to the site to express the polypeptide, whereby the disorder or condition is treated.    
     
     
         23 . The method of  claim 22 , wherein the disorder or condition is a proliferative or angiogenic disorder or condition.  
     
     
         24 . The method of  claim 22 , wherein the functional IL12 polypeptide comprises a first subunit and a second subunit.  
     
     
         25 . The method of  claim 24 , wherein the first subunit comprises: 
 (a) a polypeptide encoded by a nucleic acid molecule of SEQ ID NO: 1 or 5;    (b) a polypeptide encoded by a nucleic acid substantially identical to SEQ ID NO:1 or 5;    (c) a polypeptide having an amino acid sequence set forth as SEQ ID NO:2 or 6;    (d) a polypeptide having an amino acid sequence substantially identical to SEQ ID NO:2 or 6; or    (e) a polypeptide that is a biological equivalent of SEQ ID NO:2 or 6.    
     
     
         26 . The method of  claim 24 , wherein the second subunit comprises: 
 (a) a polypeptide encoded by a nucleic acid molecule of SEQ ID NO:3 or 7;    (b) a polypeptide encoded by a nucleic acid substantially identical to SEQ ID NO:3 or 7;    (c) a polypeptide having an amino acid sequence set forth as SEQ ID NO:4 or 8;    (d) a polypeptide having an amino acid sequence substantially identical to SEQ ID NO:4 or 8; or    (e) a polypeptide that is a biological equivalent of SEQ ID NO:4 or 8.    
     
     
         27 . The method of  claim 22 , wherein the polynucleotide comprises: 
 (a) a nucleotide sequence set forth as SEQ ID NO:1, 3, 5, or 7; or    (b) a nucleotide sequence that is substantially identical to SEQ ID NO:l, 3, 5, or 7.    
     
     
         28 . The method of  claim 22 , wherein the heat-inducible promoter comprises an hsp70 promoter.  
     
     
         29 . The method of  claim 28 , wherein the hsp70 promoter comprises a human hsp70B promoter.  
     
     
         30 . The method of  claim 29 , wherein the hsp70B promoter comprises: 
 (a) the nucleotide sequence set forth as SEQ ID NO:13, or functional portion thereof; or    (b) a nucleotide sequence substantially identical to SEQ ID NO:13.    
     
     
         31 . The method of  claim 22 , wherein the site in need of treatment is a tumor.  
     
     
         32 . The method of  claim 31 , wherein treating the disorder comprises promoting tumor regression.  
     
     
         33 . The method of  claim 32 , wherein the tumor regression is characterized by an immune response and an anti-angiogenic response.  
     
     
         34 . The method of  claim 33 , further comprising providing radiotherapy to the tumor, whereby anti-tumor activity is enhanced compared to anti-tumor activity in response to radiotherapy alone.