Superficial Conjunctival Cells from Dupilumab-treated Atopic Dermatitis Patients with Ocular Adverse Events Display a Transcriptomic Psoriasis Signature

Description

Dupilumab has demonstrated efficacy in the treatment of atopic dermatitis (AD). However, a subset of patients experiences ocular adverse events (OAE), including conjunctivitis and dry eye syndrome, the pathological mechanisms of which are still unknown Study design We conducted a bicentric clinical study between Toulouse and Nantes Hospitals. It is ancillary to the clinical study in Toulouse from 2017 and the multicentric DUPIOEIL study from 2019 in Toulouse and Nantes. This study was conducted in accordance with the declaration of Helsinki. Informed consent was obtained from all participants at M0. Population All patients who needed to be treated with dupilumab were aged over 18 years. Dermatology and ophthalmology examinations were performed on the same day, before treatment introduction. Four months after the beginning of dupilumab treatment (M4) the same ophthalmologist (MC in Toulouse and MW in Nantes) examined the patients again. We excluded patients who could not be consulted before treatment or at M4. The ophthalmological exams concerns the ocular surface status (slit lamp exam, Shirmer test I, Oxford scale and conjunctival impression). Occurrence of OAE was defined as an aggravation of the Oxford scale or development of a conjunctivitis at M4 compared to M0. At M4, we divided the patients in two groups: one group who developed OAE (OAE+) and a control group with patients presenting a comparable ophthalmological exam as before dupilumab treatment (OAE-). Conjunctival cells collection At M0 and M4 examinations, a conjunctival impression was performed in routine examination to analyze ocular surface. The conjunctival impression was collected by the same practitioner (MC in Toulouse and MW in Nantes) for all patients at M0 and M4. After an instillation of anesthetic drop (oxybuprocain, Théa Pharma, Clermont-Ferrand, France), a conjunctival impression device (Eyeprim, Opia technologies SAS, Paris, France) was used to collect cells from temporal superior quadrant at 2 mm from the limbus. The sampling was immediately preserved in liquid nitrogen and then stocked à - 80°C for later evaluation. RNA extraction and isolation Total RNA was directly extracted from the membranes with Qiagen MicroRNA extraction kit according to the manufacturer's instructions (Qiagen S.A.S., Cortaboeuf, France). Reverse transcriptase was performed using the Invitrogen Superscript III VILO kit according to the manufacturer’s recommendations (ThermoFisher Scientific, Villebon-sur-Yvette, France). RNA quantity and quality were determined using QBIT system (Thermofisher Scientific) and Agilent (Agilent, CA). Selected samples had an RNA integrity number above 8 and RNA concentration above 25 ng/µl. Microarray Analysis A full transcriptome microarray analysis was performed using the human Clariom™ S Assay platform (Affymetrix, Thermo Fisher Scientific,Waltham, MA, USA) at the Genotoul Get Biopuces facility (Toulouse, France). Five ng per sample was processed.

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