molecular signatures order the potency of topically applied anti inflammatory drugs in atopic dermatitis patients

Molecular signatures order the potency of topically applied anti-inflammatory drugs inatopic dermatitis patients Emma Guttman-Yassky, MD PhD, Benjamin Ungar, BA, Kunal Malik, BA, Daniel

Molecular signatures order the potency of topically applied anti-inflammatory drugs in

atopic dermatitis patients

Emma Guttman-Yassky, MD PhD, Benjamin Ungar, BA, Kunal Malik, BA, Daniel

Dickstein, BA, Maria Suprun, MPH, Yeriel D Estrada, BS, Hui Xu, MSc, Xiangyu

Peng, MSc, Margeaux Oliva, BA, Dan Todd, MSc, Tord Labuda, PhD, Mayte

Suarez-Farinas, PhD, Robert Bissonnette, MD

DOI: 10.1016/j.jaci.2017.01.027

Reference: YMAI 12660

To appear in: Journal of Allergy and Clinical Immunology

Received Date: 20 September 2016

Revised Date: 21 December 2016

Accepted Date: 5 January 2017

Please cite this article as: Guttman-Yassky E, Ungar B, Malik K, Dickstein D, Suprun M, Estrada YD, Xu

H, Peng X, Oliva M, Todd D, Labuda T, Suarez-Farinas M, Bissonnette R, Molecular signatures order

the potency of topically applied anti-inflammatory drugs in atopic dermatitis patients, Journal of Allergy and Clinical Immunology (2017), doi: 10.1016/j.jaci.2017.01.027.

This is a PDF file of an unedited manuscript that has been accepted for publication As a service to our customers we are providing this early version of the manuscript The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Table E1 Adverse Events

Number of AEs per patient Frequency (%) Adverse Event

Frequency (%)

1 13 (59.09) Allergic rhinitis 1 (4.55)

2 5 (22.73) Common cold 2 (9.09)

3 2 (9.09) Conjunctivitis 1 (4.55)

4 1 (4.55) Constipation 1 (4.55)

5 1 (4.55) Cut of the finger 1 (4.55)

BD 0.05%** 1 (4.55) Headache 1 (4.55) None 21 (95.45) Hot flush 1 (4.55)

Yes 0 (0) Insect bite L forearm 1 (4.55)

No 22 (100) Intermittent Coughing 1 (4.55)

Related 0 (0) Intermittent headache 1 (4.55) Not Related 22 (100) Nausea 2 (9.09)

** Diagnosis is not related to treatment

Adverse Events/AE; Biopsy/Bx; Left/L; Related/rel.; betamethasone diproprionate 0.05%/BD 0.05%

Total Sign Score

Veh - Pim 1% 8 vs 1 1.379 (0.0307) 0.634 Veh - BD 0.05% 8 vs 1 2.69 (<0.0001) 0.634 Veh - CP 0.05% 8 vs 1 2.828 (<0.0001) 0.634 Pim 1% - BD 0.05% 8 vs 1 1.31 (0.04) 0.634 Pim 1% - CP 0.05% 8 vs 1 1.448 (0.0234) 0.634

BD 0.05% - CP 0.05% 8 vs 1 0.138 (0.828) 0.634 Veh - Pim 1% 15 vs 1 0.724 (0.255) 0.634 Veh - BD 0.05% 15 vs 1 2.724 (<0.0001) 0.634 Veh - CP 0.05% 15 vs 1 3.379 (<0.0001) 0.634 Pim 1% - BD 0.05% 15 vs 1 2 (0.0018) 0.634 Pim 1% - CP 0.05% 15 vs 1 2.655 (<0.0001) 0.634

BD 0.05% - CP 0.05% 15 vs 1 0.655 (0.303) 0.634

Target Area Assessment

Veh - Pim 1% 8 vs 1 0.517 (0.02) 0.221 Veh - BD 0.05% 8 vs 1 0.759 (0.0007) 0.221 Veh - CP 0.05% 8 vs 1 0.793 (0.0004) 0.221 Pim 1% - BD 0.05% 8 vs 1 0.242 (0.275) 0.221 Pim 1% - CP 0.05% 8 vs 1 0.276 (0.212) 0.221

BD 0.05% - CP 0.05% 8 vs 1 0.034 (0.877) 0.221 Veh - Pim 1% 15 vs 1 0.31 (0.161) 0.221 Veh - BD 0.05% 15 vs 1 1 (<0.0001) 0.221 Veh - CP 0.05% 15 vs 1 1.069 (<0.0001) 0.221 Pim 1% - BD 0.05% 15 vs 1 0.69 (0.002) 0.221 Pim 1% - CP 0.05% 15 vs 1 0.759 (0.0007) 0.221

BD 0.05% - CP 0.05% 15 vs 1 0.069 (0.756) 0.221

B

A

BD 0.5% - CP 0.5% 8 vs 1 0.463 (0.938) 5.985 Veh - Pim 1% 15 vs 1 2.189 (0.715) 5.985 Veh - BD 0.05% 15 vs 1 14.880 (0.0136) 5.985 Veh - CP 0.05% 15 vs 1 13.110 (0.0295) 5.985 Pim 1% - BD 0.05% 15 vs 1 12.691 (0.0351) 5.985 Pim 1% - CP 0.05% 15 vs 1 10.921 (0.0694) 5.985

BD 0.05% - CP 0.05% 15 vs 1 -1.770 (0.768) 5.985

Vehicle/Veh; Pimecrolimus 1%/Pim 1%; Betamethasone Diproprionate 0.05%/BD

0.05%; Clobetasol Proprionate 0.05%/CP 0.05%; Standard error of the mean/SEM

Spearman Correlation with

Total Sign Score Reduction

Total Area Assessment/TAA

Spearman Correlation with Epidermal Thickness

CD11c 0.704 <0.001 FCεRI 0.595 <0.001 CD3 0.568 <0.001 IL-13 0.558 <0.001 DC-LAMP 0.548 <0.001 TSS 0.533 <0.001 CXCL10 0.53 <0.001 S100A7 0.524 <0.001 CCL22 0.522 <0.001 K16 0.511 <0.001 MMP12 0.5 <0.001 TAA 0.494 <0.001 Langerin 0.491 <0.001 CXCL9 0.475 <0.001 CXCL1 0.474 <0.001 CCL26 0.471 <0.001 S100A8 0.47 <0.001 CCL17 0.448 <0.001 PI3 0.444 <0.001 IL-22 0.434 <0.001 CCL18 0.431 <0.001 S100A12 0.424 <0.001 S100A9 0.413 <0.001 CD206 0.41 <0.001 IFNγ 0.345 <0.001 IL-12/23p40 0.333 <0.001 IL-31 0.315 <0.01 IL-23p19 0.261 <0.01 IL-17A 0.244 <0.01 FLG -0.211 <0.05 TSC22 -0.509 <0.001 TXNIP -0.512 <0.001 FKBP5 -0.563 <0.001 LOR -0.609 <0.001

Total Sign Score/TSS; Total Area Assessment/TAA

Table E4 Antibodies used for immunohistochemical analyses

FceR1 Thermo Scientific CRA1(AER37) IgG2b, kappa 1:100

a

Antibodies used in this experiment were mouse monoclonal unless stated otherwise

Table E6 Mean trans-epidermal water loss values across treatment days

Treatment Visit Day Mean TEWL Value (g/m 2 /h) Standard Error

Table E7 Change from baseline (day 1) for each investigational product

Total Sign Score

Interval

Vehicle 8 vs 1 -0.897 (0.047) (-1.781, -0.012) Vehicle 15 vs 1 -2.207 (<0.0001) (-3.092, -1.322) Pimecrolimus 1% 8 vs 1 -2.276 (<0.0001) (-3.161, -1.391) Pimecrolimus 1% 15 vs 1 -2.931 (<0.0001) (-3.816, -2.046) Betamethasone 0.05% 8 vs 1 -3.586 (<0.0001) (-4.471, -2.702) Betamethasone 0.05% 15 vs 1 -4.931 (<0.0001) (-5.816, -4.046) Clobetasol 0.05% 8 vs 1 -3.724 (<0.0001) (-4.609, -2.840) Clobetasol 0.05% 15 vs 1 -5.586 (<0.0001) (-6.471, -4.702)

Target Area Assessment

Interval

Vehicle 8 vs 1 -0.173 (0.027) (-0.481, 0.135) Vehicle 15 vs 1 -0.587 (0.0002) (-0.895, -0.279) Pimecrolimus 1% 8 vs 1 -0.691 (<0.0001) (-0.998, -0.383) Pimecrolimus 1% 15 vs 1 -0.897 (<0.0001) (-1.205, -0.590) Betamethasone 0.05% 8 vs 1 -0.932 (<0.0001) (-1.240, -0.625) Betamethasone 0.05% 15 vs 1 -1.588 (<0.0001) (-1.895, -1.280) Clobetasol 0.05% 8 vs 1 -0.967 (<0.0001) (-1.275, -0.659) Clobetasol 0.05% 15 vs 1 -1.656 (<0.0001) (-1.964, -1.349)

Trans-epidermal Water Loss

Interval

Vehicle 8 vs 1 -15.786 (0.00024) (-24.136, -7.436) Vehicle 15 vs 1 -18.957 (<0.0001) (-27.307, -10.608) Pimecrolimus 1% 8 vs 1 -19.377 (<0.0001) (-27.727, -11.027) Pimecrolimus 1% 15 vs 1 -21.146 (<0.0001) (-29.496, -12.796) Betamethasone 0.05% 8 vs 1 -29.344 (<0.0001) (-37.695, -20.993) Betamethasone 0.05% 15 vs 1 -33.837 (<0.0001) (-42.188, -25.486) Clobetasol 0.05% 8 vs 1 -29.807 (<0.0001) (-38.158, -21.457) Clobetasol 0.05% 15 vs 1 -32.067 (<0.0001) (-40.417, -23.717)

A

B

C