Primary and Exploratory Endpoint (Clinical Efficacy) Analysis
It was assumed that hives were not self-limiting and any improvement in
outcomes during the study were because of interventions. For primary
end-point analysis (i.e., UAS7), data from subjects completing the study
(n=9) were analyzed; whereas for the CU-QoL total score exploratory
end-point, an intent-to-treat (n=12) and complete responder (n=9)
analysis was performed. A repeated-measures mixed-model regression
analysis using the ‘glimmix ’ procedure was used to fit a
generalized-linear-mixed model for comparison of UAS7 and CU-QoL total
scores between visits and between responder phenotypes adjusted for
their baseline severity of disease. Correlational analysis between UAS7
and CU-QoL total scores were performed to associate clinical
improvements with improved QoL.
Secondary Endpoint
Analysis
Cell Infiltrates in Lesional and Non-Lesional Skin Biopsies.Quantitative analysis of mononuclear and mast cells observed around
postcapillary venules in lesional skin vs. non-lesional skin at visits 2
and 5 were performed to determine the effect of benralizumab on cell
infiltration as its possible therapeutic mechanism in CSU.
Peripheral Blood White Cell Counts. Relative white cell counts
in peripheral blood (eos%, basophil%) over the entire duration of the
study was performed to determine any significant associations of changes
in eos% or basophil%, due to treatment with benralizumab, with the
corresponding mean ΔUAS7 and differential gene expression in peripheral
blood pre- and post-benralizumab.
Comparison Between
Responders And Non-Responders To Benralizumab. Analysis of the effect
of benralizumab on the differential white cell count in peripheral blood
between responders and non-responders was performed to determine
effect.
Differential Gene
Expression In Blood. For differential gene expression in peripheral
blood, the RNA sequence reads were aligned to the reference genome using
the TopHat aligner 29, and reads aligning to each
known transcript were counted using Bioconductor packages for
next-generation sequencing data analysis. 30 DEGs were
analyzed in Ingenuity Pathway Analysis (IPA) platform (Qiagen) for
determination of pathway enrichment and upstream regulators.
RESULTS
Subject Characteristics. Twelve CSU and 6 non-CSU (control)
subjects were enrolled. The mean age of CSU subjects (F:M-9:3,
Black:White-2:10) was 47.3 years. The average duration (with 95% CI) of
CSU symptoms was 7.0 (0.2-18) years. Most CSU subjects were prescribed
diphenhydramine or hydroxyzine as rescue therapy. Nine subjects
completed the study. Three (25%) of the 12 subjects dropped out of the
study after the first benralizumab dose (one due to personal issues, one
lost to follow-up and one due to non-response); among them, two showed a
trend to response and one was a non-responder who had previously also
not responded to omalizumab.