Diagnostic and screening methods for atopic dermatitis

Abstract

The present disclosure provides methods for identifying agents that are candidate agents for treating atopic dermatitis. The present disclosure provides methods for diagnosing atopic dermatitis. The present disclosure provides compositions and methods for treating atopic dermatitis.

1 .- 47 . (canceled) 
     
     
         48 . A method of treating atopic dermatitis in an individual, the method comprising:
 obtaining a skin sample of the individual containing a keratinocyte;   detecting a level of a gene product of one or more of ORAI1, STIM1, and NFAT in the keratinocyte present in the skin sample, wherein an elevated level of the gene product of one or more of ORAI1, STIM1, and NFAT in the keratinocyte, compared to a normal control level, indicates that the individual has atopic dermatitis; and   treating the individual with an effective amount of an agent that reduces the level of the gene product of one or more of ORAI1, STIM1, and NFAT in the keratinocyte present in the skin sample.   
     
     
         49 . The method of  claim 48 , wherein the one or more of an ORAI1 gene product, a STIM1 gene product, and an NFAT1 gene product is an mRNA, and wherein the level of the mRNA is determined by a quantitative polymerase chain reaction. 
     
     
         50 . The method of  claim 48 , wherein the one or more of an ORAI1 gene product, a STIM1 gene product, and an NFAT1 gene product in the keratinocyte is a polypeptide. 
     
     
         51 . The method of  claim 48 , wherein said treating comprises administering a nucleic acid agent that reduces the level of ORAI1 in the keratinocyte in the individual. 
     
     
         52 . The method of  claim 48 , wherein said treating comprises administering a nucleic acid agent that reduces the level of STIM1 in the keratinocyte in the individual. 
     
     
         53 . The method of  claim 51 , wherein the nucleic acid agent is an antisense nucleic acid or a short interfering RNA. 
     
     
         54 . The method of  claim 52 , wherein the nucleic acid agent is an antisense nucleic acid or a short interfering RNA. 
     
     
         55 . The method of  claim 53 , wherein the nucleic acid agent comprises at least one modified nucleotide. 
     
     
         56 . The method of  claim 54 , wherein the nucleic acid agent comprises at least one modified nucleotide. 
     
     
         57 . The method of  claim 55 , wherein the modified nucleotide is a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, a 2′-amino-modified nucleotide, a 2″-alkyl-modified nucleotide, a morpholino nucleotide, a phosphoramidate, or a non-natural base comprising nucleotide. 
     
     
         58 . The method of  claim 48 , wherein the one or more of an ORAI1 gene product, a STIM1 gene product, and an NFAT1 gene product in the keratinocyte is determined using an immunological assay. 
     
     
         59 . The method of  claim 58 , wherein the immunological assay is an enzyme-linked immunosorbent assay. 
     
     
         60 . The method of  claim 58 , wherein the immunological assay is a radioimmunoassay. 
     
     
         61 . The method of  claim 58 , wherein the immunological assay is a protein Western Blot. 
     
     
         62 . The method of  claim 58 , wherein the immunological assay is an assay that detects a fusion partner of a fusion protein. 
     
     
         63 . The method of  claim 48 , wherein detecting the level of the one or more of an ORAI1 gene product, a STIM1 gene product, and an NFAT1 gene product in the keratinocyte is determined by assessing the level of Thymic Stromal Lymphopoietin polypeptide (TSLP) TSLP produced by the keratinocyte. 
     
     
         64 . The method of  claim 63 , wherein assessing the level of TSLP produced by the keratinocyte is carried out using a calcium-sensing dye. 
     
     
         65 . A method of detecting a gene product of one or more of an ORAI1, a STIM1, and an NFAT1 present in a skin sample of an individual, the method comprising:
 a) obtaining a keratinocyte from the skin sample of the individual;   b) assaying for an expression level of the gene product of the one or more of an ORAI1, a STIM1, and an NFAT1 in the keratinocyte present in the skin sample; and   c) detecting whether the gene product of the one or more of an ORAI1, a STIM1, and an NFAT1 is present in the keratinocyte at an expression level that is higher than the expression level of a normal control level of the gene product of one or more of an ORAI1, a STIM1, and an NFAT1 in a keratinocyte control sample.   
     
     
         66 . The method of  claim 65 , wherein the one or more of an ORAI1 gene product, a STIM1 gene product, and an NFAT1 gene product is an mRNA, and wherein the level of the mRNA is determined by a quantitative polymerase chain reaction. 
     
     
         67 . The method of  claim 65 , wherein detecting the level of the one or more of an ORAI1 gene product, a STIM1 gene product, and an NFAT1 gene product in the keratinocyte is determined by assessing the level of Thymic Stromal Lymphopoietin polypeptide (TSLP) TSLP produced by the keratinocyte.