Reagents and methods for identifying and modulating expression of genes regulated by retinoids

Abstract

This invention identifies growth-inhibitory genes induced by retinoids. The invention provides reagents and methods for identifying compounds other than retinoids that induce expression of these cellular genes. The invention also provides reagents that are recombinant mammalian cells containing recombinant expression constructs that express a reporter gene under the transcriptional control of a promoter for a gene that is regulated by retinoids, and methods for using such cells to identify compounds other than retinoids that modulate expression of these cellular genes.

1 . A method for identifying a non-retinoid compound that induces expression of a retinoid-inducible gene in a mammalian cell, comprising the steps of: 
 (a) culturing the cell in the presence and absence of the compound;    (b) assaying the cell for changes in expression of at least one cellular gene whose expression is induced by a retinoid wherein the promoter does not contain a RARE site; and    (c) identifying the compound as an inducer of retinoid-induced gene expression if expression of the cellular genes of subpart (b) is higher in the presence of the compound.    
     
     
         2 . The method of  claim 1 , wherein the cellular gene is insulin-like growth factor binding protein-3 (IGFBP-3), secreted cell adhesion protein βIG-H3, epithelial protein lost in neoplasm (EPLIN), ubiquitin-like protein FAT10, Mac-2 binding protein (Mac-2 BP), Protein C inhibitor (PCI), T cell receptor gamma, retinal oxidase, Bene, HIF-2alpha/EPAS- 1, selectin L, or proteasome activator PA28 subunit α(PA28α).  
     
     
         3 . The method of  claim 1 , wherein the cellular gene is a gene wherein expression thereof in a mammalian cell is induced by a retinoid and inhibits growth of the cell thereby.  
     
     
         4 . The method of  claim 1 , wherein the cellular gene is human insulin-like growth factor binding protein-3 (IGFBP-3), secreted cell adhesion protein βIG-H3, epithelial protein lost in neoplasm (EPLIN), ubiquitin-like protein FAT10 or proteasome activator PA28 subunit α(PA28α).  
     
     
         5 . The method of  claim 1 , wherein expression of the cellular gene is detected using an immunological reagent.  
     
     
         6 . The method of  claim 1 , wherein expression of the cellular gene is detected by assaying for an activity of the cellular gene product, wherein the cellular gene is insulin-like growth factor binding protein-3 (IGFBP-3), secreted cell adhesion protein βIG-H3, epithelial protein lost in neoplasm (EPLIN), ubiquitin-like protein FAT10, Mac-2 binding protein (Mac-2 BP), Protein C inhibitor (PCI), T cell receptor gamma, retinal oxidase, Bene, HIF-2alpha/EPAS-1, selectin L, or proteasome activator PA28 subunit α(PA28α).  
     
     
         7 . The method of  claim 1 , where expression of the cellular gene is detected by hybridization to a complementary nucleic acid.