Nucleic acid sequences controlling lung cell-specific gene expression

Abstract

An oligonucleotide which includes at least one nucleic acid sequence which binds to at least one nuclear protein found in lung cells, such as TTF-1 protein. The oligonucleotide may be contained in a vector. The at least one nuclear protein provides for lung cell-specific expression of the vector upon binding of the at least one nucleic acid sequence to the at least one nuclear protein. Such vector may also include genes encoding therapeutic agents, and may be employed for delivering genes encoding therapeutic agents to lung cells.

What is claimed is:  
     
         1 . An oligonucleotide including a nucleic acid sequence which binds a nuclear protein found in lung cells, said nucleic acid sequence which binds to a nuclear protein found in lung cells having the following structure: 
 WXNNYZ, wherein W is cytosine, guanine, or thymine; X is cytosine, thymine, or adenine; N is adenine, cytosine, guanine, thymine; Y is adenine, thymine, or guanine; and Z is guanine, adenine, or cytosine.    
     
     
         2 . The oligonucleotide of    claim 1    wherein W is cytosine.  
     
     
         3 . The oligonucleotide of    claim 1    wherein X is thymine.  
     
     
         4 . The oligonucleotide of    claim 1    wherein Y is adenine.  
     
     
         5 . The oligonucleotide of    claim 1    wherein Z is guanine.  
     
     
         6 . The oligonucleotide of    claim 1    wherein said oligonucleotide includes a nucleic acid sequence having the following structure: 
 CTNNAG.  
 
     
     
         7 . The oligonucleotide of    claim 6    wherein said nuclear protein found in lung cells is TTF-1 protein.  
     
     
         8 . The oligonucleotide of    claim 7    wherein said oligonucleotide further includes a nucleic acid sequence which binds to HNF-3 protein.  
     
     
         9 . The oligonucleotide of    claim 1    wherein said oligonucleotide includes a nucleic acid sequence having the following structure: 
 CAAG  
 
     
     
         10 . The oligonucleotide of    claim 9    wherein said nuclear protein found in lung cells is TTF-1 protein.  
     
     
         11 . A vector including at least one nucleic acid sequence which binds to at least one nuclear protein found in lung cells, said at least one nuclear protein providing for lung cell-specific expression of said vector upon binding of said at least one nucleic acid sequence to said at least one nuclear protein.  
     
     
         12 . The vector of    claim 11    wherein said at least one nucleic acid sequence which binds to said at least one nuclear protein found in lung cells is contained in the proximal promoter region of the human surfactant protein B gene as defined by the region from base −218 to base +41 of said human surfactant protein B gene.  
     
     
         13 . The vector of    claim 12    wherein said at least one nucleic acid sequence which binds to said at least one nuclear protein found in lung cells is contained in a portion of the proximal promoter region of the human surfactant protein B gene as defined by the region from base −118 to base −64 of the human surfactant protein B gene.  
     
     
         14 . The vector of    claim 11    wherein said vector includes a nucleic acid sequence which binds to TTF-1 protein.  
     
     
         15 . The vector of    claim 14    wherein said nucleic acid sequence which binds to TTF-1 protein includes a nucleic acid sequence which has the following structure: 
 WXNNYZ, wherein W is cytosine or guanine; X is cytosine, or thymine; N is adenine, cytosine, guanine, or thymine; Y is adenine, or thymine; and Z is guanine or adenine.  
 
     
     
         16 . The vector of    claim 15    wherein W is cytosine.  
     
     
         17 . The vector of    claim 15    wherein X is thymine.  
     
     
         18 . The vector of    claim 15    wherein Y is adenine.  
     
     
         19 . The vector of    claim 15    wherein Z is guanine.  
     
     
         20 . The vector of    claim 11    wherein said vector is a viral vector.  
     
     
         21 . The vector of    claim 20    wherein said vector is an adenoviral vector.  
     
     
         22 . The vector of    claim 20    wherein said vector is a retroviral vector.  
     
     
         23 . The vector of    claim 15    wherein said nucleic acid sequence has the following structure: 
 CTNNAG.  
 
     
     
         24 . The vector of    claim 14    wherein said nucleic acid sequence which binds TTF-1 protein includes a nucleic acid sequence which has the following structure: 
 CAAG.  
 
     
     
         25 . The vector of    claim 11   , and further including at least one nucleic acid sequence encoding a therapeutic agent.  
     
     
         26 . The vector of    claim 14   , and further comprising a nucleic acid sequence which binds to HNF-3 protein.  
     
     
         27 . The vector of    claim 11    wherein said at least one nucleic acid sequence which binds to said at least one nuclear protein is contained within the promoter region of the CCSP protein gene.  
     
     
         28 . The vector of    claim 11    wherein said at least one nucleic acid sequence which binds to said at least one nuclear protein is contained within the promoter region of the surfactant protein C gene.  
     
     
         29 . A method of effecting expression of a therapeutic agent in lung cells of a host, comprising: 
 administering to a host a vector including at least one nucleic acid sequence which binds to at least one nuclear protein found in lung cells, and at least one nucleic acid sequence encoding a therapeutic agent, said at least one nuclear protein providing for lung cell-specific expression of said vector upon binding of said at least one nucleic acid sequence to said at least one nuclear protein, said vector being administered in an amount effective to produce a therapeutic effect in said host.    
     
     
         30 . The method of    claim 29    wherein said at least one nucleic acid sequence which binds to said at least one nuclear protein found in lung cells is contained in the proximal promoter region of the human surfactant protein B gene as defined by the region from base −218 to +41 of said human surfactant protein B gene.  
     
     
         31 . The method of    claim 30    wherein said at least one nucleic acid sequence which binds to said at least one nuclear protein found in lung cells in contained in a portion of the proximal promoter region of the human surfactant protein B gene as defined by the region from base −118 to base −64 of the human surfactant protein B gene.  
     
     
         32 . The method of    claim 29    wherein said vector includes a nucleic acid sequence which binds to TTF-1 protein.  
     
     
         33 . The method of    claim 32    wherein said nucleic acid sequence which binds to TTF-1 protein includes a nucleic acid sequence having the following structure: 
 WXNNYZ, wherein W is cytosine or guanine; X is cytosine, or thymine; N is adenine, cytosine, guanine, or thymine; Y is adenine, or thymine; and Z is guanine or adenine.  
 
     
     
         34 . The method of    claim 33    wherein W is cytosine.  
     
     
         35 . The method of    claim 33    wherein X is thymine.  
     
     
         36 . The method of    claim 33    wherein Y is adenine.  
     
     
         37 . The method of    claim 33    wherein Z is guanine.  
     
     
         38 . The method of    claim 32    wherein said vector further includes a nucleic acid sequence which binds to HNF-3 protein.  
     
     
         39 . The method of    claim 32    wherein said vector is a viral vector.  
     
     
         40 . The method of    claim 39    wherein said vector is an adenoviral vector.  
     
     
         41 . The method of    claim 39    wherein said vector is a retroviral vector.  
     
     
         42 . The method of    claim 33    wherein said nucleic acid sequence includes a nucleic acid sequence having the following structure: 
 CTNNAG.  
 
     
     
         43 . The method of    claim 32    wherein said nucleic acid sequence includes a nucleic acid sequence having the following structure: 
 CAAG.  
 
     
     
         44 . A method of detecting cancer which has originated in the lung or thyroid, comprising: 
 obtaining a sample of cancer cells from a patient;    contacting said cancer cells with a nucleic acid sequence which includes at least one nucleic acid sequence which binds to TTF-1 protein; and    determining the binding of said nucleic acid sequence, which includes at least one nucleic acid sequence which binds to TTF-1 protein, to said cancer cells.    
     
     
         45 . A method of treating a patient suffering from a cancer which has originated in the lung, comprising: 
 administering to said patient a vector including at least one nucleic acid sequence which binds to TTF-1 protein and a negative selective marker, whereby said vector transduces said cells of said cancer which has originated in the lung; and    administering to said patient an interaction agent which interacts with the protein expressed by said negative selective marker, said interaction agent being administered in an amount effective to kill the cells of said cancer which has originated in the lung.    
     
     
         46 . The method of    claim 45    wherein said vector is a retroviral vector.  
     
     
         47 . The method of    claim 45    wherein said negative selective marker is the Herpes Simplex thymidine kinase gene.  
     
     
         48 . A method of detecting cancer which has originated in the lung, comprising: 
 obtaining a sample of cancer cells from a patient;    contacting said cancer cells with at least one antibody which recognizes an epitope of a protein selected from the group consisting of nuclear proteins found in lung cells and lung surfactant proteins; and    determining the binding of said at least one antibody which recognizes an epitope of a protein selected from the group consisting of nuclear proteins found in lung cells and lung surfactant proteins, to said cancer cells.    
     
     
         49 . An isolated polynucleotide comprising a member selected from the group consisting of: 
 (a) a polynucleotide encoding human TTF-1 protein;    (b) a polynucleotide having at least 90% homology to the polynucleotide of (a);    (c) a polynucleotide encoding a protein having at least 90% homology to human TTF-1 protein;    (d) a polynucleotide capable of hybridizing to any one of polynucleotides (a), (b), or (c); and    (e) a polynucleotide fragment of any one of polynucleotides (a), (b), (c), or (d).    
     
     
         50 . The polynucleotide of    claim 49    which comprises nucleotides 199 to 569 and 1,533 to 2,372 of the sequence shown in FIG. 39.  
     
     
         51 . A method of detecting cancer which has originated in the lung or thyroid, comprising: 
 obtaining a sample of cancer cells from a patient;    contacting nucleic acids of said cancer cells with one or more of polynucleotides (a), (b), (c), (d), or (e) of    claim 49   ; and    determining the binding of said one or more polynucleotides to said nucleic acids of said cancer cells.