Spotlight on a Short-Time Treatment with the IL-4/IL-13 Receptor Blocker in Patients with CRSwNP: microRNAs Modulations and Preliminary Clinical Evidence

Abstract

Already used for the treatment of some allergic and inflammatory diseases, such as asthma or atopic dermatitis, dupilumab has also been approved as add-on therapy for patients with CRSwNP, and it could represent the keystone to reducing the remission time as well as to improve healing and quality of life. On the other hand, the role of miRNAs as potential biomarkers of immune modulation is emerging. We analyzed the effects of a short-time treatment with dupilumab in patients with CRSwNP, analyzing the immune response modification as well as miRNAs modulations. First, in this early observation stage, all patients experienced remarkable improvement and were clinically stable. Indeed, we observed a significant decrease in CD4+ T cells and a significant reduction in total IgE (p < 0.05) and serum IL-8 levels (p < 0.01), indicating a reduction in the general inflammatory condition. In addition, we analyzed a panel of about 200 circulating miRNAs. After treatment, we noted a significant downregulation of hsa-mir-25-3p (p-value = 0.02415) and hsa-mir-185-5p (p-value = 0.04547), two miRNAs involved in the proliferation, inflammation, and dug-resistance, in accordance with the clinical status of patients. All these preliminary data aimed to identify new biomarkers of prognosis, identifiable with non-invasive procedures for patients. Further, these patients are still under observation, and others with different levels of responsiveness to treatment need to be enrolled to increase the statistical data.

Keywords: T cells; anti-IL-4/IL-13 receptor; antibody therapeutics; dupilumab; interleukins; miRNAs; nasal polyposis.

Figure 1

Clinical parameters pre (T0) and post (T3) treatment with Dupilumab. Patients were evaluated with regards to canonical diagnostic/prognostic scores, such as SNOT-22 (panel A), NPS (panel B), and VAS (panel C). All parameters were expressed as means ± SD. Differences between two groups (T3 vs. T0) were analyzed by paired two-tailed t-test; * p < 0.05, ** p < 0.01.

Figure 2

IgE (A) and IL-8 (B) levels in serum of patients with CRSwNP before (T0) and after (T1, T2, T3) treatment with Dupilumab; dotted line indicates the reference value threshold. (C) Flow cytometry assay of circulating CD3⁺/CD4⁺ T cells in blood of the most representative patient (Pt1) during the observation; the percentage of CD3⁺/CD4⁺ T cells was reported in the Q2 square of each plot panel. (D) Percentage of CD3⁺/CD4⁺ T cells in the overall patients at the indicated timepoints; data are reported as the mean ± s.e.m. Data were analyzed by ANOVA for repeated measures, followed by Bonferroni’s multiple comparison test; * p < 0.05, ** p < 0.01.

Figure 3

Serum miRNAs profiling of patients at baseline (T0) and after treatment (T3). Heatmap displaying the results of a hierarchical cluster analysis (HCA) conducted on the patients’ serum levels of a 98 miRNAs panel at baseline and after the end of the treatment. Log2 fold expression values of reference are reported in the upper bars.

Figure 4

(A) Volcano plot of the 98 serum miRNAs analyzed by qRT-PCR (T3 vs. T0). Significant down-regulated miRNAs are localized in the upper-left section (dots colored green). (B) Serum miR-25-3p and miR-185-5p levels at T3 compared to baseline. Values are represented in whiskers plots as fold expression. Statistical significance is indicated at the top of the graph. * p ≤ 0.05.