Th17 differentiation markers for acne and uses thereof

Abstract

A method is described for using ROR gamma t or ROR alpha to diagnose acne and/or to screen inhibitors of Th17 differentiation. Specifically described are methods of inhibiting ROR gamma t or ROR alpha and use of the screened inhibitors in acne treatment.

1 . An acne marker, the marker comprising DNA or mRNA encoding ROR gamma t, or a corresponding protein. 
     
     
         2 . An acne marker, the marker comprising DNA or mRNA encoding ROR alpha, or a corresponding protein. 
     
     
         3 . An acne marker, the marker comprising DNA or mRNA encoding ROR gamma t or a corresponding protein, DNA or mRNA encoding ROR alpha or a corresponding protein and/or at least one marker selected from the group consisting of IL-6, IL-17A, IL-17F, IL21, IL-22, IL-23A, IL-26, and CCL20. 
     
     
         4 . A method for diagnosing acne, the method comprising the following steps of:
 a) detecting a level of expression of the marker of  claim 1 , and/or at least one markers selected from the group consisting of IL-17A, IL-17F, IL-22, and CCL20 in a sample from an individual,   b) detecting a level of expression of the marker of  claim 1 , and/or at least one marker selected from the group consisting of IL-17A, IL-17F, IL-22, and CCL20 in a sample from a healthy individual,   c) comparing a difference in level of expression of at least one marker and for which the level of expression is significantly higher than the level of expression in the healthy individual; and   d) wherein an overexpression of at least one of the markers of step c) is an indicator of acne, thus diagnosing acne.   
     
     
         5 . A method for diagnosing acne, the method comprising the following steps of:
 a) detecting a level of expression of the marker of  claim 1  in a sample from an individual,   b) detecting a level of expression of the marker of  claim 1  in a sample from a healthy individual,   c) comparing a difference in level of expression of at least one marker and for which the level of expression is significantly higher than the level of expression in the healthy individual; and   d) wherein an overexpression of at least one of the markers of step c) is an indicator of acne, thus diagnosing acne.   
     
     
         6 . A method for monitoring progression or variation of acne, the method comprising the following steps of:
 a) taking a biological sample from an individual,   b) analyzing a level of expression of at least one proposed markers, and/or at least one marker selected from the group consisting of IL-6, IL-17A, IL-17F, IL-22, and CCL20 in a sample and in which a variation in expression of at least one of the markers is an indicator of the progression of acne.   
     
     
         7 . A method for monitoring the efficacy of a treatment intended for treating acne, the method comprising the following steps of:
 a) administering a desired treatment to an individual identified as having one or more symptoms of acne,   b) taking a biological sample from the individual,   c) analyzing a level of expression of the marker of  claim 1  and/or at least one marker selected from the group consisting of IL-17A, IL-17F, IL-22, and CCL20, in which a variation in expression of at least one of the markers is an indicator of efficacy of the treatment of acne.   
     
     
         8 . An in vitro screening method of Th17 cells differentiation inhibitors, the method comprising determining the capacity of a candidate to inhibit or down regulate expression and/or to inhibit biological function, including transactivation activity, of the marker of  claim 1 . 
     
     
         9 . An in vitro screening method of Th17 cells differentiation inhibitors for identifying drug candidates, the method comprising the following steps of:
 a) collecting at least two biological samples: wherein one sample mimics an acne lesion, and the other sample mimics a healthy condition;   b) contacting at least one sample or a mixture of samples with one or more drug candidates to be tested;   c) detecting expression or biological function of at least one proposed markers, and/or at least one expression marker selected from the group consisting of: IL-17A, IL-17F, IL-22, and CCL20 in the biological sample or mixture obtained in b);   d) selecting drug candidates that inhibit expression or biological function of IL-17A, IL-17F, IL-22, CCL20 measured in said sample or mixture obtained in b) and comparing the levels with a sample not mixed with the drug candidate.   
     
     
         10 . An in vitro screening method of Th17 cell inhibitors for drug candidate identification, the method comprising the following steps of:
 a) collecting at least two biological samples: wherein one sample mimics an acne lesion, and the other sample mimics a healthy condition;   b) contacting at least one sample or a mixture of samples with one or more drug candidates to be tested;   c) detecting expression or biological function of the marker as defined in  claim 1  in the biological sample or mixture obtained in step b); and   d) selecting drug candidates that are capable of inhibiting expression or biological function of the marker as defined in  claim 1  measured in said sample or mixture obtained in step b) and comparing levels or biological function with a sample not mixed with the drug candidate.   
     
     
         11 . A method of preparing a composition for treating acne and/or acne associated disorders, the method comprising preparing the composition using inhibitors identified by screening methods as defined in  claim 10 . 
     
     
         12 . A method of preparing a composition for treating acne or acne associated disorders, the method comprising preparing the composition using inhibitors of markers of  claim 1  identified by screening methods for preparing the composition selected from the group consisting of N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-benzenesulfonamide, 2 oxysterol (oxygenated sterols), especially 24S-hydroxycholesterol 24(S), 25-epoxycholesterol and 7-oxygenated sterols, Methyl 2-cyano-3,12-dioxooleana-1,9(11)dien-28-oate or Bardoxolone methyl, (8S,9R,10R,13R,14S,16R,17R)-17-[(E,2R)-2,6-dihydroxy-6-methyl-3-oxohept-4-en-2-yl]-2,16dihydroxy-4,4,9,13,14-pentamethyl-8,10,12,15,16,17-hexahydro-7H-cyclopenta[a]phenanthrene-3,11-dione, 5-(4-chlorophenyl)-6-ethylpyrimidine-2,4-diamine, gamma-D-glutamyl-L-tryptophan, 8-hydroxy-3-methyl-3,4-dihydro-2H-benzo[a]anthracene-1,7,12-trione,5,7dihydroxy-2-(4-hydroxyphenyl)-4-oxo-4H-chromen-3-olate, methyl-N-[4-(trifluoromethyl)phenyl]-1,2-oxazole-4-carboxamide or Leflunomide, and N-[(E)-(3-methylphenyl)methylideneamino]-6-morpholin-4-yl-2-(2-pyridin-2-ylethoxy)pyrimidin-4-amine.