Methods for diagnosing atopic dermatitis using gene classifiers
Disclosed herein are methods of detecting an altered gene expression levels in a subject suspected of having atopic dermatitis. Further described herein are methods of treating atopic dermatitis in a subject having an exhibiting an altered gene expression level.
1. A method for preparing nucleic acids from a skin sample useful for detecting atopic dermatitis in a human subject, comprising:
a) isolating nucleic acids from a skin sample obtained from the subject, where the skin sample is obtained by applying an adhesive patch to the subject's skin;
b) detecting gene expression levels of one or more target genes, comprising at least IL-31, by contacting the isolated nucleic acids with probes that recognize nucleic acids expressed by the target genes and detecting binding of the probes;
c) identifying that expression levels of the one or more target genes are increased at least two-fold compared to a normal control sample,
d) diagnosing the subject with the at least two-fold increase in gene expression as having atopic dermatitis; and
e) administering an inhibitor of one or more of IL-13, PDE4 and IL-31 to the diagnosed subject.
2. The method of claim 1 , wherein the one or more target genes are used in a gene classifier.
3. The method of claim 2 , wherein the gene classifier comprises IL-13 or TSLP.
4. The method of claim 3 , wherein the expression level comprises an upregulated expression level of IL-13 or TSLP, relative to one or more controls.
5. The method of claim 2 , wherein the gene classifier comprises IL-13 and TSLP.
6. The method of claim 5 , wherein the expression levels of IL-13, or TSLP is increased relative to one or more controls.
7. The method of claim 6 , wherein b) comprises detecting the expression levels of IL-13, IL-31, and TSLP by contacting the isolated nucleic acids with a set of probes that recognizes IL-13, IL-31, and TSLP, and detecting binding between IL-13, IL-31, and TSLP and the set of probes.
8. The method of claim 1 , further comprising:
detecting the expression level of IL-13R, IL-4R, IL-17, IL-22, IL-23A, CXCL9, CXCL10, CXCL11, S100A7, S100A8, S100A9, CCL17, CCL18, CCL19, CCL26, CCL27, or NOS2, by contacting the isolated nucleic acids with an additional probe that recognizes IL-13R, IL-4R, IL-17, IL-22, IL-23A, CXCL9, CXCL10, CXCL11, S100A7, S100A8, S100A9, CCL17, CCL18, CCL19, CCL26, CCL27, or NOS2, and detecting binding between IL-13R, IL-4R, IL-17, IL-22, IL-23A, CXCL9, CXCL10, CXCL11, S100A7, S100A8, S100A9, CCL17, CCL18, CCL19, CCL26, CCL27, or NOS2 and the additional probe.
9. The method of claim 8 , wherein IL-13 is used in a first gene classifier, and one or more of the following genes is used in a second gene classifier: IL-13R, IL-4R, IL-17, IL-22, IL-23A, CXCL9, CXCL10, CXCL11, S100A7, S100A8, S100A9, CCL17, CCL18, CCL19, CCL26, CCL27, or NOS2.
10. The method of claim 9 , wherein two or more of the following genes is used in the second gene classifier: IL-13R, IL-4R, IL-17, IL-22, IL-23A, CXCL9, CXCL10, CXCL11, S100A7, S100A8, S100A9, CCL17, CCL18, CCL19, CCL26, CCL27, or NOS2.
11. The method of claim 10 , wherein b) comprises detecting the expression levels of the two or more genes from the second gene classifier by contacting the isolated nucleic acids with a set of probes that recognizes the two or more genes from the second gene classifier, and detecting binding between the two or more genes from the second gene classifier and the set of probes.
12. The method of claim 8 , wherein IL-31 is used in a first gene classifier, and one or more of the following genes is used in a second gene classifier: IL-13R, IL-4R, IL-17, IL-22, CXCL9, CXCL10, CXCL11, S100A7, S100A8, S100A9, CCL17, CCL18, CCL19, CCL26, CCL27, or NOS2.
13. The method of claim 1 , wherein the sample comprises cells from the stratum corneum.
14. The method of claim 1 , wherein an amount of the nucleic acids isolated from the skin sample is from about 100 picograms to about 1 microgram.
15. The method of claim 1 , wherein the inhibitor comprises an antibody.
16. The method of claim 1 , wherein the inhibitor comprises one or more of lebrikizumab, tralokinumab, crisaborole, and nemolizumab.