MicroRNA profiling in women with migraine: effects of CGRP-targeting treatment

Abstract

Background: Migraine lacks biomarkers that can trace the biological pathways of the disease and predict the effectiveness of treatments. Monoclonal antibodies targeting calcitonin gene-related peptide pathway - including erenumab - offer the opportunity of investigating potential migraine biomarkers due to their specific mechanism of action in preventing both episodic (EM) and chronic (CM) migraine. Our study aims at evaluating the expression levels of circulating microRNAs (miRNAs) according to migraine type, before and after treatment with erenumab and based on treatment response, in order to identify miRNAs with potential role as epigenetic biomarkers.

Methods: The study included women aged 25-50 years with EM or CM treated with erenumab according to clinical indications. MiRNAs expression levels were assessed before (baseline) and after a 16-week treatment with erenumab, 140 mg every four weeks (post-treatment). An extensive miRNAs profiling was performed by qRT-PCR in small, pooled groups of ≤ 8 women each, classified according to migraine frequency (EM and CM) and the degree of response to erenumab. The expression levels of selected miRNAs were also validated using single miRNA assays in each woman with EM and CM.

Results: During the study, 36 women with migraine (19 with EM and 17 with CM) out of 40 who were initially screened, performed the assessment of miRNA expression at baseline and post-treatment, Erenumab treatment significantly improved migraine burden in both EM and CM. MiRNA profiling revealed differential expression levels of a wide set of miRNAs (hsa-let-7d-3p, hsa-miR-106b-3p, hsa-miR-122-5p, hsa-miR-143-3p, hsa-miR-144-3p, hsa-miR-16-5p, hsa-miR-181a-5p, hsa-miR-221-3p, hsa-miR-25-3p, hsa-miR-29b-2-5p, hsa-miR-326, miR-363-3p, hsa-miR-424-5p, hsa-miR-485-3p, hsa-miR-532-5p, hsa-miR-543, hsa-miR-629-5p, hsa-miR-660-5p, hsa-miR-92a-3p) depending on treatment response. Among them, single miRNA assays confirmed the progressive decrease of hsa-miR-143-3p expression levels in relation to increasing response to erenumab in women with EM (7 with low, 6 with medium, and 6 with high response; p = 0.02). Additionally, single assays showed higher hsa-miR-34a-5p and hsa-miR-382-5p expression levels at baseline in women with CM compared with those with EM (p = 0.0002 and p = 0.0007, respectively), as well as their expression level decrease in women with CM from baseline to follow-up (p = 0.04 and p = 0.02, respectively).

Conclusions: Our study suggests that targeting the CGRP pathway in migraine changes the expression levels of certain miRNAs. These miRNA levels are linked to the levels of response to CGRP receptor blockage. Future research challenges include assigning specific functions to the modulated miRNAs to unravel pathways modulated by the disease and the treatment.

Trial registration: The study was registered in clinicaltrials.gov with code NCT04659226 and in the Novartis database with code CAMG334AIT05T.

Keywords: Biomarkers; Epigenetics; Erenumab; MicroRNA; Migraine prevention; Monoclonal antibodies.

Fig. 1

Flowchart of womens’ inclusion

Fig. 2

Mean change in efficacy outcomes of erenumab in women with episodic migraine (a, b) and chronic migraine (c, d) in the present study. *p < 0.05 **p < 0.001. MMDs indicates monthly migraine days; MHDs, monthly headache days; MMedDs, monthly medication days; MMedInts, monthly medication intakes; MIDAS, Migraine Impact and Disability Assessment Scale; HIT-6, Headache Impact Test-6; BDI, Beck Depression Inventory; ASC-12, Allodynia Symptom Checklist-12; PSQI, Pittsburgh Sleep Quality Index

Fig. 3

Change in levels of microRNAs at baseline (a) and post-treatment (b) in women with episodic migraine according to erenumab response. Change in levels of microRNAs at baseline (c) and post-treatment (d) in women with chronic migraine according to erenumab response. *p < 0.05; significant p-values are in bold. RQ indicates relative quantification

Fig. 4

Boxplot showing miRNA expression levels in individual women with episodic or chronic migraine according to erenumab response, validated using single miRNA assays: miR-363-3p in women with episodic migraine at baseline (a), miR-143-3p in women with episodic migraine post-treatment (b), miR-29b-2-5p in women with chronic migraine at baseline (c) and miR-326 in women with chronic migraine post-treatment (d). Statistically significant p-values are reported

Fig. 5

Boxplot showing miR-34a-5p and miR-382-5p expression levels in women with episodic or chronic migraine, validated using single miRNA assays. a,b miR-34a-5p and miR-382-5p expression levels in women with chronic migraine compared to those with episodic migraine. c,d miR-34a-5p and miR-382-5p expression levels in women with chronic migraine post-treatment. e MiR-382-5p expression levels in women with episodic migraine post-treatment. Only statistically significant p-values are reported

Fig. 6

Results of the Receiver-Operating Characteristic curve analysis of miR-34a-5p and miR-382-5p to discriminate women with chronic migraine from those with episodic migraine and to assess the difference from baseline to post-treatment in each of the two groups. The most promising combination is highlighted in bold and shown in the plot. Abbreviations: Area Under the Curve (AUC), sensitivity (SE), specificity (SP), accuracy (ACC), optimal cutoff (Opt Cut-off)