Methods for diagnosing and managing treatment of atopic dermatitis

Patent metadata

Field	Value
Publication number	US-11376261-B2
Application number	US-201816224143-A
Country	US
Kind code	B2
Filing date	Dec 18, 2018
Priority date	Dec 18, 2017
Publication date	Jul 5, 2022
Grant date	Jul 5, 2022

Abstract

Provided herein is a method of diagnosing and treating a subject suffering from atopic dermatitis (AD), the method comprising: (a) obtaining a skin biopsy from a subject suspected of suffering from AD; (b) determining a level of RNA expression in the skin biopsy of genes selected from the 89ADGES gene panel; (c) comparing the determined level of RNA expression of the selected genes to the level of RNA expression of the selected genes in a reference sample comprising RNA expression products from normal healthy skin cells; (d) diagnosing the subject as suffering from AD when specific genes are up-regulated compared to the reference sample and when specific genes are down-regulated compared to the reference sample; and (e) treating the subject with a therapy effective for the treatment of AD. Methods of managing treatment of a subject suffering from AD are also provided.

First claim

We claim:

1. A method of diagnosing and treating a subject suffering from atopic dermatitis (AD), the method comprising:
(a) obtaining a skin biopsy from a subject suspected of suffering from AD;
(b) determining a level of RNA expression in the skin biopsy of a plurality of genes comprising SERPINB4, AKR1B10, SERPINB3, S100A7, DEFB4A, RRM2, CCL18, PI3, COL6A6, KRT16, CCL22, NCAPG, TMPRSS4, KRT6, SELE, GALNT6, PGF, CCL17, APOBEC3A, CBLC, CD1B, IGL, CDSN, BTC, Clorf46, Clorf68, ASPRV1, PSOR1C2, IL1F7, POF1B, LOR, SLURP1, MSMB, CST6, ELMODJ, FABP7, CARD18, CTSL2, GREM1, GPLD1, HBA, ARG1, ANXA9, DKK2, KLK5, LYVE1, PSG7, AQP9, CPA3, GPD1, HSD11B1, ARHGAP18, EREG, CLDN23, FLG, SCEL, and CHP2;
(c) comparing said determined level of RNA expression of the plurality of genes to the level of expression of the plurality of genes in a reference sample comprising RNA expression products from normal healthy skin cells;
(d) diagnosing the subject as suffering from AD when SERPINB4, AKR1B10, SERPINB3, S100A7, DEFB4A, RR1112, CCL18, PI3, COL6A6, KRT16, CCL22, NCAPG, TMPRSS4, KRT6, SELE, GALNT6, PGF, CCL17, APOBEC3A, CBLC, CD1B, and IGL are up-regulated compared to the reference sample and when CDSN, BTC, Clorf46, Clorf68, ASPRV1, PSOR1C2, IL1F7, POF1B, LOR, SLURP1, MSMB, CST6, ELMOD1, FABP7, CARD18, CTSL2, GREM1, GPLD1, HBA, ARG1, ANXA9, DKK2, KLK5, LYVE1, PSG7, AQP9, CPA3, GPD1, HSD11B1, IGJ, ARHGAP18, EREG, CLDN23, FLG, SCEL, and CHP2 are down-regulated compared to the reference sample; and
(e) treating the diagnosed subject with one or more therapeutic agents selected from the group consisting of corticosteroids, betamethasone, tacrolimus, pimecrolimus, narrow-band UVB, PDE4 inhibitors, tofacitinib, dupilumab, and nemolizumab.

2. The method of claim 1 , wherein determining RNA expression levels comprises molecular methods selected from the group consisting of quantitative RT-PCR, microarray, and RNA-seq.

3. The method of claim 1 , wherein determining a level of RNA expression in the skin biopsy comprises:
(a) isolating RNA from the skin biopsy;
(b) using the isolated RNA to create cRNA;
(c) labeling the cRNA with a fluorescent dye; and
(d) hybridizing the labeled cRNA to a microarray.

4. A method of diagnosing and treating a subject suffering from atopic dermatitis (AD), the method comprising:
(a) obtaining a skin biopsy from a subject suspected of suffering from AD;
(b) generating a skin biopsy transcriptional profile comprising RNA expression levels of a plurality of genes comprising SERPINB4, AKR1B10, SERPINB3, S100A7, DEFB4A, RRM2, CCL18, PI3, COL6A6, KRT16, CCL22, NCAPG, TMPRSS4, KRT6, SELE, GALNT6, PGF, CCL17, APOBEC3A, CBLC, CD1B, IGL, CDSN, BTC, Clorf46, Clorf68, ASPRV1, PSOR1C2, IL1F7, POF1B, LOR, SLURP1, MSMB, CST6, ELMODJ, FABP7, CARD18, CTSL2, GREM1, GPLD1, HBA, ARG1, ANXA9, DKK2, KLK5, LYVE1, PSG7, AQP9, CPA3, GPD1, HSD11B1, IGJ, ARHGAP18, EREG, CLDN23, FLG, SCEL, and CHP2;
(c) comparing the skin biopsy transcriptional profile to an AD signature transcriptional profile of the plurality of genes;
(d) diagnosing the subject as suffering from AD when the skin biopsy transcriptional profile and the AD signature transcriptional profile are at least 25% concordant; and
(e) treating the diagnosed subject with one or more therapeutic agents selected from the group consisting of corticosteroids, betamethasone, tacrolimus, pimecrolimus, narrow-band UVB, PDE4 inhibitors, tofacitinib, dupilumab, and nemolizumab.

5. The method of claim 4 , wherein generating a skin biopsy transcriptional profile comprises:
(a) determining RNA expression levels in the skin biopsy for the selected genes; and
(b) comparing the RNA expression levels of the selected genes in the skin biopsy to control RNA expression levels from normal healthy skin cells to generate the skin biopsy transcriptional profile.

6. The method of claim 4 , wherein determining RNA expression levels comprises molecular methods selected from the group consisting of quantitative RT-PCR, microarray, and RNA-seq.

7. The method of claim 4 , wherein determining RNA expression levels in the skin biopsy comprises:
(a) isolating RNA from the skin biopsy;
(b) using the isolated RNA to create cRNA;
(c) labeling the cRNA with a fluorescent dye; and
(d) hybridizing the labeled cRNA to a microarray.

8. The method of claim 1 , wherein determining the level of RNA expression in the skin biopsy of the plurality of genes of step (b) further comprises determining the level of RNA expression of one or more of S100A8, RGS1, KYNU, TYMP, IL27RA, GPR171, GZMB, EHBP1L1, TACC3, LCE2B, FADS1, CRCT1, SERPINB7, RNASE7, GAL, SLC46A2, HBB, CORIN, SCGB2A1, OGN, LOC100130476, LGR5, C15orf48, FAR2, HPGDS, SLIT2, SFRP2, MUC15, OMD, LOX, or DIO2; and
wherein step (d) further comprises diagnosing the subject as suffering from AD when one or more of S100A8, RGS1, KYNU, TYMP, IL27RA, GPR171, GZMB, EHBPIL1, or TACC3 is upregulated compared to the reference sample; and/or diagnosing the subject as suffering from AD when one or more of LCE2B, FADS1, CRCT1, SERPINB7, RNASE7, GAL, SLC46A2, HBB, CORIN, SCGB2A1, OGN, LOC100130476, LGR5, C15orf48, FAR2, HPGDS, SLIT2, SFRP2, MUC15, OMD, LOX or DIO2 is down-regulated compared to the reference sample.

9. The method of claim 4 , wherein the plurality of genes further comprises one or more of S100A8, RGS1, KYNU, TYMP, IL27RA, GPR171, GZMB, EHBPIL1, TACC3, LCE2B, FADS1, CRCT1, SERPINB7, RNASE7, GAL, SLC46A2, HBB, CORIN, SCGB2A1, OGN, LOC100130476, LGR5, C15orf48, FAR2, HPGDS, SLIT2, SFRP2, MUC15, OMD, LOX, or DIO2.

Assignees

• Univ Cincinnati
• Cincinnati Childrens Hospital Medical Center
• Childrens Hospital Med Ct

Inventors

• Ghosh Debajyoti
• Mersha Tesfaye
• Bernstein Jonathan A

Classifications

• C12Q1/6883

  for diseases caused by alterations of genetic material · CPC title

• G16B30/10

  Sequence alignment; Homology search · CPC title

• C12Q2600/158

  Expression markers · CPC title

• G16B20/00

  ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations · CPC title

• C12Q2600/106

  Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism · CPC title