Detection for HPV-Induced invasive cancers and their precursor lesions with invasive potential

Abstract

This invention provides a method of detecting an HPV-induced invasive cancer or precursor lesion thereof associated with tumor suppressor lung cancer 1 (TSLC1) in a subject in need thereof. The method comprises contacting a cell component of a test cell of the subject with a reagent that detects the level of the cell component in the test cell and determining a modification in the level of the cell component in the test cell as compared with a comparable healthy cell. The cell component indicates the level of TSLC1 in the cell, and a decrease in the level of TSLC1 indicates the presence of an HPV-induced invasive cancer or precursor lesion.

1 . A method of detecting an HPV-induced invasive cancer or precursor lesion thereof associated with tumor suppressor lung cancer 1 (TSLC1) in a subject in need thereof, the method comprising contacting a cell component of a test cell of the subject with a reagent that detects the level of the cell component in the test cell and determining a modification in the level of the cell component in the test cell as compared with a comparable healthy cell, wherein the cell component indicates the level of TSLC1 in the cell and wherein a decrease in the level of TSLC1 indicates the presence of an HPV-induced invasive cancer or precursor lesion thereof. 
     
     
         2 . A method according to  claim 1 , wherein the HPV-induced invasive cancer or precursor lesion thereof is invasive cervical cancer or a premalignant cervical lesion with invasive potential. 
     
     
         3 . A method according to  claim 1 , wherein the HPV-induced invasive cancer is a high-risk HPV-induced invasive cancer. 
     
     
         4 . A method according to  claim 2 , wherein the HPV-induced invasive cancer is a high-risk HPV-induced invasive cancer. 
     
     
         5 . A method according to  claim 3 , wherein the cell component is a nucleic acid associated with production of TSLC1 polypeptide, the reagent targets the nucleic acid in the test cell, and the nucleic acid encodes the TSLC1 and regulatory regions. 
     
     
         6 . A method according to  claim 5 , wherein the nucleic acid is RNA. 
     
     
         7 . A method according to  claim 6 , wherein the reagent is a methylation sensitive restriction endonuclease. 
     
     
         8 . A method according to  claim 5 , wherein the reagent is a restriction endonuclease. 
     
     
         9 . A method according to  claim 6 , wherein the reagent is a nucleic acid probe or primer that binds to the nucleic acid, having a detectable label. 
     
     
         10 . A method according to  claim 8 , wherein the nucleic acid probe has a one of the following nucleotide sequences:
 a) a polynucleotide sequence capable of hybridizing under stringent conditions to the 5′ regulatory region or the coding region of the TSCL1 sequence as set forth in  FIG. 1 ;   b) a polynucleotide sequence having at least 70% identity to the polynucleotide of a);   c) a polynucleotide sequence complementary to the polynucleotide sequence of a); or   d) a polynucleotide sequence comprising at least 15 bases of a polynucleotide sequence of a) or b).   
     
     
         11 . A method according to  claim 10 , wherein the cell component is a polypeptide and the reagent targets the polypeptide in the test cell, and wherein the polypeptide is TSLC1 and the reagent is an anti-TSLC1 antibody. 
     
     
         12 . A method according to  claim 10 , wherein the method of detecting evaluates the methylation status of the TSLC1 promoter. 
     
     
         13 . A method according to  claim 10 , wherein the target cellular component is a protein. 
     
     
         14 . A method according to  claim 10 , wherein the reagent is an anti-TSLC1 antibody. 
     
     
         15 . A method of treating HPV-induced invasive cancers and their precursor lesions associated with modification of TSLC1 production in cells in a subject afflicted with such a cancer or lesion, the method comprising contacting cells of the subject with a therapeutically effective amount of a reagent that increases TSLC1 level in the cells of the subject. 
     
     
         16 . A method according to  claim 14 , wherein the reagent includes a polynucleotide sequence comprising a TSLC1 sense polynucleotide sequence, and wherein the polynucleotide is a native, unmethylated TSLC1 sense sequence. 
     
     
         17 . A method according to  claim 15 , wherein a nonmethylatable analog is substituted for cytidine within the TSLC1 sense sequence, and wherein the nonmethylatable analog preferably being is 5-azacytadine. 
     
     
         18 . A method according to  claim 16 , wherein the polynucleotide sequence is contained in an expression vector, and the expression vector is a plasmid, a viral particle or a phage. 
     
     
         19 . A molecular diagnostic marker for detection of progression to invasiveness of HPV-induced premalignant lesions associated with tumor suppressor lung cancer 1 (TSLC1) and for detection of future metastatic potential of HPV-induced premalignant lesions and carcinomas associated with tumor suppressor lung cancer 1 (TSLC1), wherein the marker indicates TSLC1 promoter methylation, expression of mRNA associated with production of TSLC1 polypeptide, or both. 
     
     
         20 . A kit for use in a method of detecting HPV-induced invasive cancers and their precursor associated with tumor suppressor lung cancer 1 (TSLC1) in test cells of a subject, the kit comprising means to collect test cells and the molecular diagnostic marker of  claim 19 . 
     
     
         21 . A kit for use in a method of detecting HPV-induced invasive cancers and their precursor lesions associated with tumor suppressor lung cancer 1 (TSLC1) in test cells of a subject, the kit comprising primers and probes capable of hybridizing to TSLC1 nucleotide sequence of  FIG. 1 , TSLC1 antibodies, or methylation sensitive restriction enzymes recognizing the sequence as described in  FIG. 1 . 
     
     
         22 . A kit according to  claim 20 , wherein the test cells are cervical cells.