Expression Inhibitor of Inflammation Promoting Factor, Screening Method for Active Ingredient Thereof, Expression Cassette Useful for Said Method, Diagnostic Agent and Diagnosis Method

Abstract

Provided are an expression inhibitor of an inflammation promoting factor based on the discovery of a new factor influencing the expression amount/level of an inflammation promoting factor, and a development tool therefor. Provided are also a diagnostic agent and a diagnosis method for immune diseases, inflammatory diseases, painful conditions and similar. More specifically provided are: an expression inhibitor of an inflammation promoting factor containing at least one kind of inhibitor selected from the group consisting of RBMS2 expression inhibitor and RBMS2 function inhibitor; a screening method using as an indicator the expression or the function of RBMS2; an expression cassette useful for said method; as well as a diagnostic agent containing a product detection agent for RBMS2 gene expression and disease detection method using as an indicator RBMS2 gene expression amount/level.

1 . A reagent for screening for an active ingredient for an inhibitor of the expression of an inflammation promoting factor, containing at least one component selected from the group consisting of an expression cassette containing a RBMS2 gene expression regulation region and a gene arranged in such a manner that the expression thereof can be regulated by the region, a vector carrying the expression cassette and a cell harboring the vector. 
     
     
         2 . A method for screening for an active ingredient for an inhibitor of the expression of an inflammation promoting factor by employing at least one item selected from the group consisting of items (i) to (iii) as an index in the presence of a test substance:
 (i) the expression amount/level of a gene of which the expression can be regulated by a RBMS2 gene expression regulation region;   (ii) the binding amount/level of RBMS2 to RNA containing an AU-rich element; and   (iii) the amount/level of mRNA containing an AU-rich element in a 3′-UTR thereof or the amount/level of a protein derived from the mRNA in a RBMS2-overexpressing cell.   
     
     
         3 . The screening method according to  claim 2 , wherein, when the value of the index in the presence of a test substance is smaller than the value of the index in the absence of the test substance, the test substance is selected as an active ingredient for an inhibitor of the expression of an inflammation promoting factor. 
     
     
         4 . The screening method according to  claim 2 , wherein the AU-rich element is an AU-rich element derived from at least one type of mRNA selected from the group consisting of IL-6 mRNA, COX-2 mRNA, IL-8 mRNA, IL-1β mRNA, TNF-α mRNA, MMP1 mRNA, IL-24 mRNA and c-Myc mRNA. 
     
     
         5 . The screening method according to  claim 2 , wherein the method includes steps (a1) to (c1):
 (a1) bringing an expression system which contains an expression cassette containing a RBMS2 gene expression regulation region and a gene arranged in such a manner that the expression thereof can be regulated by the region into contact with a test substance;   (b1) measuring, as an expression amount/level of interest, the amount/level of the gene expressed in the expression system that has been contacted with the test substance and then comparing the expression amount/level of interest with a control expression amount/level that is the expression amount/level of the gene in an expression system that has not been contacted with the test substance; and   (c1) selecting the test substance as an active ingredient for an inhibitor of the expression of an inflammation promoting factor when the expression amount/level of interest is smaller than the control expression amount/level.   
     
     
         6 . The screening method according to  claim 5 , wherein the expression system is a cell. 
     
     
         7 . The screening method according to  claim 5 , wherein the gene is a reporter gene. 
     
     
         8 . The screening method according to, wherein the method includes steps (a2) to (c2):
 (a2) bringing RNA containing an AU-rich element into contact with RBMS2 in the presence of a test substance;   (b2) measuring the binding amount/level between the RNA and the RBMS2 which are contacted with each other in the presence of a test substance as a binding amount/level of interest, and then comparing the binding amount/level of interest with a control binding amount/level that is the binding amount/level between the RNA and the RBMS2 which are contacted with each other in the absence of the test substance; and   (c2) selecting the test substance as an active ingredient for an inhibitor of the expression of an inflammation promoting factor when the binding amount/level of interest is smaller than the control binding amount/level.   
     
     
         9 . The screening method according to  claim 2 , wherein the method includes steps (a3) to (c3):
 (a3) bringing a cell which contains mRNA containing an AU-rich element in a 3′-UTR thereof and in which RBMS2 is overexpressed into contact with a test substance;   (b3) measuring the amount/level of the mRNA or a protein derived from the mRNA in the cell that has been contacted with the test substance as an amount/level of interest, and then comparing the amount/level of interest with a control amount/level that is the amount/level of the mRNA or a protein derived from the mRNA in a cell that is not contacted with the test substance; and   (c3) selecting the test substance as an active ingredient for an inhibitor of the expression of an inflammation promoting factor when the amount/level of interest is smaller than the control amount/level.   
     
     
         10 . The screening method according to  claim 9 , wherein the mRNA contains an ORF of a reporter protein. 
     
     
         11 . An inhibitor of the expression of an inflammation promoting factor containing at least one component selected from the group consisting of a RBMS2 expression inhibitor and a RBMS2 function inhibitor. 
     
     
         12 . The inhibitor of the expression of an inflammation promoting factor according to  claim 11 , wherein the RBMS2 expression inhibitor contains at least one RBMS2 expression inhibitor selected from the group consisting of RBMS2-specific siRNA, RBMS2-specific miRNA, a RBMS2-specific antisense nucleic acid, expression vectors for these components and IL-10. 
     
     
         13 . The inhibitor of the expression of an inflammation promoting factor according to  claim 11 , wherein the inflammation promoting factor of which the expression is to be inhibited is at least one component selected from the group consisting of IL-6, COX-2, IL-8, IL-1β, TNF-α, MMP1, IL-24 and c-Myc. 
     
     
         14 . The inhibitor of the expression of an inflammation promoting factor according to  claim 11 , wherein the inhibitor is used as a prophylactic or therapeutic agent for at least one disease selected from the group consisting of immune diseases, inflammatory diseases and painful diseases. 
     
     
         15 . A diagnostic agent for a disease which can be developed or worsened by an inflammation promoting factor, which contains a RBMS2 gene expression product detecting agent. 
     
     
         16 . The diagnostic agent according to  claim 15 , wherein the disease is at least one disease selected from the group consisting of immune diseases, inflammatory diseases and painful diseases. 
     
     
         17 . The diagnostic agent according to  claim 15 , wherein the inflammation promoting factor is at least one component selected from the group consisting of IL-6, COX-2, IL-1β, IL-8, TNF-α, MMP1, IL-24 and c-Myc. 
     
     
         18 - 20 . (canceled)