Methods of screening for cell proliferation or neoplastic disorders

Abstract

The invention relates to methods and compositions for identifying subjects having, or predisposed to having, a neoplastic or cell proliferation or neoplastic disorder. The methods are applicable to any type of tissue sample and can be conducted on otherwise normal tissue.

What is claimed is: 
     
         1 . A method of determining predisposition of a subject to developing a neoplastic or cell proliferation disorder comprising determining the ratio of undifferentiated to differentiated cells in the same or different sample from the subject, wherein the ratio of undifferentiated to differentiated cells, as compared to a reference ratio, is indicative of a predisposition for developing a neoplastic or cell proliferation disorder. 
     
     
         2 . The method of  claim 1 , further comprising identifying a cell displaying abnormal expression of a target gene that directly or indirectly results from loss of imprinting. 
     
     
         3 . The method of  claim 2 , wherein the target gene is selected from H19 or IGF2. 
     
     
         4 . The method of  claim 2 , wherein the target gene is selected from the group consisting of Igf1R, IRS-1, IRS-2, PI3K, Akt, p70S6 kinase, FOXO, GSK3, MDM2, mTOR, Cyclin D1, c-Myc, Shc, Grb2, SOS, Ras, Raf, MEK, Erk, and MAPK gene. 
     
     
         5 . The method of  claim 2 , wherein the method comprises analyzing the biological sample for a change in the methylation status of a target gene, or a polymorphism thereof. 
     
     
         6 . The method of  claim 5 , wherein the change in methylation is hypomethylation. 
     
     
         7 . The method of  claim 5 , wherein the method comprises analyzing the biological sample for hypomethylation of both a DMR of the H19 gene and a DMR of the IGF2 gene. 
     
     
         8 . The method of  claim 3 , wherein the reference ratio is generated from tissue obtained from a subject comprising cells displaying normal imprinting of at least one of the H19 gene and the IGF2 gene. 
     
     
         9 . The method of  claim 1 , wherein determining a change in the balance or ratio of undifferentiated to differentiated cells in the sample comprises identifying a biomarker associated with a differentiated or undifferentiated cell. 
     
     
         10 . The method of  claim 9 , wherein the biomarker is selected from the group consisting of Shh (Sonic hedgehog), Tcf4, Lef1, Twist, EphB2, EphB3, Hes1, Notch1, Hoxa9, Dkk1, Tle6, Tcf3, Bmi1, Kit, Musashi1 (Msi1), Cdx1, Hes5, Oct4, Ki-67, □-catenin, Noggin, BMP4, PTEN (phosphorylated PTEN), Akt (phosphorylated Akt), Villin, Aminopeptidase N (anpep), Sucrase isomaltase (SI), Ephrin-B1 (EfnB1), Cdx2, Crip, Apoa1, Aldh1b1, Calb3, Dgat1, Dgat2, Clu, Hephaestin, Gas1, Ihh (Indian hedgehog), Intrinsic factor B12 receptor, IFABP, and KLF4. 
     
     
         11 . The method of  claim 1 , wherein determining the ratio of undifferentiated to differentiated cells in the sample comprises:
 a) imaging the sample using immunohistochemical identification of biomarker molecules specifically associated with a differentiated or undifferentiated cell population;   b) imaging the sample using standard microscopy and distinguishing differentiated from undifferentiated cells using morphologic measurements;   c) imaging the sample using immunohistochemical identification of proliferation antigens and their distribution within colonic crypts;   d) imaging the sample using immunoflourescent identification of molecules specific to a biomarker associated with a differentiated or undifferentiated cell population;   e) measuring RNA levels;   f) measuring gene expression;   g) whole genome expression analyses; or   h) allele specific expression.   
     
     
         12 . The method of  claim 1 , wherein the cells are epithelial cells. 
     
     
         13 . The method of  claim 12 , wherein the epithelial cells are obtained from a rectal Pap test. 
     
     
         14 . The method of  claim 12 , wherein the epithelial cells are obtained from intestinal tissue. 
     
     
         15 . The method of  claim 14 , wherein the intestinal tissue is obtained from the colon. 
     
     
         16 . The method of  claim 14 , wherein the epithelial cells are obtained from the lumen of the intestinal tissue. 
     
     
         17 . The method of  claim 16 , wherein the epithelial cells are obtained from the crypts of the lumen. 
     
     
         18 . The method of  claim 1 , wherein the cell proliferation or neoplastic disorder is associated with a solid tumor. 
     
     
         19 . The method of  claim 18 , wherein the solid tumor is an adenoma. 
     
     
         20 . The method of  claim 19 , wherein the adenoma is colorectal cancer. 
     
     
         21 . The method of  claim 20  wherein the subject is not previously known to have a colorectal neoplasm. 
     
     
         22 . The method of  claim 1 , further comprising correlating the ratio derived from the subject with the subject's family genetic history. 
     
     
         23 . The method of  claim 1 , wherein the subject is subjected to additional tests selected from the group consisting of chest X-rays, colorectal examinations, endoscopic examination, MRI, CAT scanning, gallium scanning, and barium imaging. 
     
     
         24 . The method of  claim 1 , wherein the subject is a human. 
     
     
         25 . The method of  claim 1 , wherein the cancer is pancreatic cancer.