Research & Innovation
Publications
BEATS – MIGRAINE: Biomarker Establishment of Altered Targets in Skin for Migraine
PMU Authors
Janine Zwicklhuber, Martin Wolf, Rodolphe Poupardin, Lisa Maria Jakob, Stefan Leis, Daniela Weber, Barbara Kofler, Elisabeth Russe, Dirk Strunk, Sarah Hochmann
All Authors
Janine Zwicklhuber, Martin Wolf, Rodolphe Poupardin, Lisa Maria Jakob, Stefan Leis, Daniela Weber, Barbara Kofler, Alena Rückl, Mark Rinnerthaler, Christof Regl, Astrid Obermayer, Elisabeth Russe, Dirk Strunk, Sarah Hochmann
Abstract
Objective
Migraine affects approximately one in five women under 50, making it the 2nd leading cause of disability worldwide. Despite its high burden, diagnosis remains solely dependent on clinical criteria due to the lack of reliable biomarkers. This project aims to address the existing gap by identifying a novel biomarker signature using skin biopsies and extracellular vesicles (EVs) derived from blood plasma.
Methods
Skin biopsies and blood samples were collected from migraine patients and matched migraine-free controls. Isolated primary keratinocytes and fibroblasts were characterized via flow cytometry (FCM) and skin sections were analyzed by immunohistochemistry (IHC). Plasma EVs were isolated using size exclusion chromatography and characterized per MISEV standards, including nanoparticle tracking analysis (NTA), FCM, transmission electron microscopy (TEM) and Western blotting (WB). Proteomic comparison was performed using TMT-based mass spectrometry. Mitochondrial activity and lipid droplet content in primary cells was assessed by IHC and FCM. Ongoing analyses include qPCR, next-generation sequencing and metabolomics.
Results
To date, a total of 66 skin biopsies from migraine patients and 93 from migraine-free controls have been collected, yielding 34 keratinocyte and 45 fibroblast primary cell lines. IHC confirmed the expression of sensory receptors OR1L1, OR5W2 and TRPM8 in both groups. Functional analysis of primary fibroblasts using MitoTracker and BODIPY revealed group-specific differences in mitochondrial activity and lipid droplet-associated pathways, as determined by FCM and IHC. EV characterization by NTA, FCM and TEM confirmed particle concentration, size distribution and vesicular morphology. WB demonstrated enrichment of the EV tetraspanin markers CD9, CD63 and CD81 in the EV⁺ fractions, while ApoA1 was restricted to the EV⁻ fractions, indicating successful depletion of lipoprotein contaminants. Comparative proteomic analysis of EVs (n=10/group) showed upregulation in immune, vascular, and immunoglobulin-related proteins in migraine.
Conclusions
The established workflows for primary cell isolation and multimodal EV characterization provide a strong foundation for the identification of migraine-associated biomarkers. The presence of cutaneous sensory receptors supports their potential involvement in migraine pathophysiology. Moreover, successful proteomic profiling of lipoprotein-depleted EVs offers a promising avenue towards the development of diagnostic tools and novel therapeutic targets for refractory migraine.
Keywords
Biomarker, MIGRAINE, extracellular vesicles (EVs)