Expression biomarkers of eczema

Transkript

1 Expression biomarkers of eczema Kilian Eyerich, MD, PhD Center of Allergy and Environment COST meeting Bern,

2 First approach: protein expression analysis in situ Advantage: very few artificial components Disadvantage: only a snap-shot; usually requires a hypothesis Used techniques: immunohistochemistry, in situ topoproteom analysis Classical immunohistochemistry: comparison of Atopy Patch Tests to pollen and epicutaneous patch tests to nickel ,5 2, 1,5 1,,5, CD3 CD8 NPT APT + APT - AE + APT - AE - CL CD45RO CD4 CL 3,5 3, CD25 CL CL 2,5 2, 1,5 1,,5, CL Eyerich K et al, Int Arch Allergy Clin Immunol 27 COST meeting Bern,

3 IgE IgE NPT APT + APT - AE + APT - AE FCeRI Ratio IFN/IL-5 Ratio IFN-g/IL-5 cells/mm² CD1a BDCA2 Epidermis CD68 CL CL CL cells/mm² 1,2 1,,8,6,4,2, , 2,5 2, 1,5 1,,5, CD1b BDCA2 Dermis CD64 CL CL COST meeting CL CL Bern, CL CL,1, IL-5 IFN-g 24h 48h 72h 96h Time point CL Ratio NPT Ratio APT NPT APT + APT - AE + APT - AE -

4 DAPI Melt IgE FceRI

5 First approach: protein expression analysis Advantage: very few artificial components Disadvantage: only a snap-shot; usually requires a hypothesis Used techniques: immunohistochemistry, in situ topoproteom analysis In situ topoproteom analysis: comparison of Atopy Patch Tests to pollen and epicutaneous patch tests to nickel SEE Epidermo-dermal Compartment Atopy Nickel Patch Epicutane t-test Epitope m ± SD m ± SD [p-value] 6838 ± ± 3168,5 BCL2 <.. CCR ± ± 14859,8185..> CD ± ± 474,2314 <.. CD11a ± ± 3132,7531..> CD ± ± 4686,2574..> CD ± ± 14438,98 <.. CD ± ± 2889,985..> CD ± ± 1635,8858 <.. CD1a 8495 ± ± 1427,273..> CD ± ± 7581,3771 <.. CD ± ± 9919,4486 <.. CD ± ± 1118,3237..> CD ± ± 15121,6337..> CD ± ± 2145,1343 <.. CD ± ± 638,733..> CD ± ± 1839,5413 <.. CD ± ± 6175,9189..> CD ± ± 3354,623..> CD ± ± 139,515 <.. CD ± ± 12334,4119 <.. CD ± ± 6855,273 <.. CD45RA 7246 ± ± 1861,912..> CD45RO 7368 ± ± 449,3221 <.. CD49f 7629 ± ± 1893,7818..> CD ± ± 738,662 <.. CD ± ± 352,64 <.. CD ± ± 136,62 <.. CD ± ± 514,2656 <.. CD ± ± 5384,9767 <.. CD62E 2239 ± ± 397,1278..> CD ± ± 1523,5365..> CD ± ± 3229,2138 <.. CD ± ± 9247,35..> CD8 416 ± ± 2362,221 <.. CLA 5621 ± ± 2966,5185..> Collagen IV ± ± 591,6178 <.. pan-keratin 4493 ± ± 18974,983 <.. Keratin ± ± 13292,3544..> Keratin ± ± 22562,575 <.. HLA-DQ 1626 ± ± 7692,4517..> HLA-DR 371 ± ± 621,5473..> Ki ± ± 452,8753 <.. MBP 2731 ± ± 1412,7425 <.. MPO 2317 ± ± 2288,2159 <.. PI 4983 ± ± 16231,446 <.. Raptiva 918 ± ± 9548,398 <.. SMA 3264 ± ± 2558,948 <.. TIA1 191 ± ± 4152,236 <.. Vimentin 2457 ± ± 8424,929 <.. COST meeting Bern,

6 COST meeting Bern,

7 APT NPT CD3 CD8 TIA-1 CD8 TIA-1 COST meeting Bern,

8 Combinatorial molecular phenotype (CMP) CD45/Bcl-2 a b c Combinatorial molecular phenotype (CMP) CLA/CD44/CD45 a b c NPT to nickel d e f d e f g h i g h i APT to pollen j k l j k l Eyerich K et al, Exp Dermatol 28 COST meeting Bern,

9 No. No. TIA1 CD13 BCL2 CLA CD68 HLA-DQ CD3 HLA-DR CD45 CD44 Epidermis CD45 CD68 CD71 CD2 CD3 CD52 CD29 CD7 CD58 CD34 CD45RA CD45RO CD138 CD38 53 * * * * * * 1 1 * * * 1 * 1 * * * * * * * 23 ± ± Keratin 1 Ki67 Klinik und Poliklinik für Dermatologie und Allergologie/ Zentrum Allergie und Umwelt Combinatorial phenotype motifs significantly higher expressed in NPT Nickel Epicutane Atopy Patch Gap between the cohorts Ratio mni-map max(m) t-test # m ± sd m ± sd [PEN] [PEN] [PEN] [p-value] 1 * 1 * * * * ± ± # * 1 * * 1 * ± ± * * * * * 7314 ± ± * * * * ± ± * * 1 * * 2454 ± ± * 1 * * * 63 ± ± Combinatorial phenotype motifs significantly higher expressed in APT Nickel Epicutane Atopy Patch Gap between the cohorts Ratio mni-map max(m) t-test # m ± sd m ± sd [PEN] [PEN] [PEN] [p-value] 7 1 * 1 * * * * 1 * * * * * * * * * * * * * 198 ± ± # 1 * * * * * * * * * * * * * * * * * 943 ± ± * 1 * 1 * * * 1 * * * * * * * * * * * * 781 ± ± * 1 * * 1 * * * * * * * * * * * * * * * * 1181 ± ± * 1 1 * 1 * * * * * * * * * * * * * * * 533 ± ± * 1 * * 1 * * * 1 * * * * * * * * * * * * 52 ± ± # * * * * * * * * * * * * 1 1 * * * * * * * 94 ± ± * 1 * * 1 1 * * * * * * * * * * * * * * * 548 ± ± * 1 * * 1 * * * * 1 * * * * * * * * * * * 416 ± ± * 1 * * 1 1 * * * 1 * * * * * * * * * * * 341 ± ± * * * * * * 1 * * * * * * * 1 * * * * * * 36 ± ± * * * * * * 1 * * * * 1 * * 1 * * * * * * 24 ± ± # 1 1 * * * * * * * * 1 * * * * * * * * * * 54 ± ± * * * * * 1 * * * * * * * * * * * * * 6 ± ± * * * * * * * 1 * * * * * * * * * * * 7 ± ± * * * * * 1 * * * * * * * * * * * * * * 215 ± ± * 1 1 * * * * * * * * * * * 1 * * * * * 76 ± ± * * * * * * * * * * * * 1 * * * * 1 * * * 21 ± ± * * * * * * * * * * * * * * 1 * * * * 31 ± ± * * * * * * * * * * * 1 1 * * * * 1 * * * 13 ± ± * * 1 1 * * * * * * * * * * * * * * * * * 42 ± ± * * * * * * * * * * * * * 1 * * * * * 25 ± ± * * * * * * 1 1 * * * * * 1 * * * * * * * 4 ± ± COST meeting Bern, * * * * * * * * * * * * * * 1 * 1 * * 15 ± 498 ±

10 Second approach: RNA expression analysis in situ Advantage: whole genome approach does not require a hypothesis Disadvantage: system is not robust (patients have to be selected very carefully) Used techniques: comparative whole genome expression arrays of NPT to nickel (n=1) and atopic eczema (n=6)/ nummular eczema (n=8) COST meeting Bern,

11 The heterogeneity of eczema Quaranta M et al, Sci Transl Med accepted COST meeting Bern,

12 The heterogeneity of eczema Significant in naturally occuring eczema only Significant in both Significant in inducible eczema only Immunology IL-1b AIM2 Numerous chemokines (CCL2/3/7/8/17/19/26, CXCL1/2/8/9/1/11) Hyluronic acid/ extracellular matrix In total 54 immune related genes Quaranta M et al, Sci Transl Med accepted COST meeting Bern,

13 The possibility to investigate two inflammatory skin diseases in parallel 6 3 SCORAD PASI Eyerich S et al, N Engl J Med 211 COST meeting Bern,

14 Diagnostic translation Quaranta M et al, Sci Transl Med accepted COST meeting Bern,

15 Thank you very much! Department for dermatology and allergy Biederstein, Munich Director: Johannes Ring Natalie Garzorz, Felix Laufer Claudia Traidl-Hoffmann, Rüdiger Hein ZAUM Center of allergy and environment, Munich Director: Carsten Schmidt-Weber Stefanie Eyerich Natalie Garzorz, Martina Mattii, Jana Sänger Jeroen Buters, Heidrun Behrendt Take home messages Possibilities to identify biomarkers of eczema subtypes are protein as well as RNA expression analyses Conventional immunohistochemistry identifies differences between APT and NPT regarding cytokine profiles and IgE In situ topoproteome analysis demonstrates more cytotoxicity in NPT as compared to APT, while more naive/ premature and more activated T cells infiltrate APT lesions A hypothesis-free investigation of induced versus naturally occurring eczema indicates new possible target genes belonging to the epidermal compartment, in particular LCE molecules Helmholtz Center Institute for computational biology, Munich Director: Fabian Theis Bettina Knapp Laboratorio di Immunologia (IDI-IRCCS), Rome Director: Andrea Cavani Francesca Nasorri, Teresa Carbone, Cristina Albanesi, Claudia Scarponi, Gianna Zambruno, Francesca Cianfarani Imperial College, London Stephen Durham, Fan Chung King s College, London Venu Pullabhatla Department for dermatology, allergology and venereology, Kiel Stephan Weidinger, Hans-Jörg Baurecht, Elke Rodriguez Financial support Bayerische Akademie der Wissenschaften Kommission Klinische Forschung A comprehensive approach within a strong community is promising and addresses urgent medical needs COST meeting Bern,