dSTORM recording of EVs

EV Biogenese
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Monitoring of EV uptake in HEK cells

TFF Set-up

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Extracellular vesicles (EVs) are small membrane particles, typically measuring between 50 and 200 nanometres in diameter. They are released by a variety of cell types. By carrying diverse cargo, including proteins, lipids, and microRNAs, EVs can trigger and regulate a wide variety of biological processes. We are developing new strategies to identify biomarkers based on improved EV phenotyping. This will ensure the valid characterisation of EV populations in different biofluids and enable innovative biomarker- and functional studies. To this end, we use and develop cutting-edge methods for EV purification and characterisation. These include, among others, size-exclusion and anion-exchange chromatography, as well as fully automated tangential flow filtration, for the scalable and reliable purification of EVs. For subsequent characterisation at the level of individual vesicles, we use microscopic techniques such as dSTORM super-resolution imaging with the NanoImager (ONI), and high-throughput diffraction-limited fluorescence and interferometric imaging with the ExoView R100 platform (Unchained Labs). These purified and characterized EV populations are then employed in various biomarker studies or functional assays.

EV-based feedback loop in lung cancer
Donzelli et al., J Extracell Vesicles. 2021

EVs from liquid biopsies as biomarkers
Saul et al., Trillium 2025

microRNA-based stratification method

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MiRNAs are a class of small non-coding RNAs that contribute to the post-transcriptional regulation of gene expression. We have identified a new function of miR-574-5p. It is based on an RNA decoy for CUG RNA-binding protein 1 (CUGBP1) and inhibits its function as a repressor of microsomal prostaglandin E synthase 1 (mPGES-1), a key enzyme in prostaglandin biosynthesis. Increased intracellular miR-574-5p expression is directly associated with increased synthesis of prostaglandin E2 (PGE2), an important proinflammatory lipid mediator. The newly discovered association between miR-574-5p and PGE2 qualifies miR-574-5p as a minimally invasive biomarker to select cancer patients likely to respond to pharmacologic inhibition of PGE2. We investigate whether paracrine mechanisms of miR-574-5p play a crucial role in tumor development. To this end, we analyze whether miR-574-5p is transferred within sEV between cancer cells and cells of the tumor microenvironment to influence tumor progression. We have chosen lung cancer and neuroblastoma as comparable tumor model systems. Both tumor types are PGE2-dependent but differ in the cellular site of PGE2 synthesis The miR-574-5p/CUGBP1 decoy regulates PGE2-biosynthesis intracellularly. High intracellular miR-574-5p induces PGE2-biosynthesis. PGE2 triggers the secretion of miR-574-5p in sEV. In recipient AC cells, sEV-derived miR-574-5p activates TLR7/8 signaling, which leads to decreased miR-574-5p, mPGES-1 and PGE2-levels.

EBC collection

EBC EVs

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Exhaled Breath Condensate (EBC) is emerging as a powerful non-invasive liquid biopsy that captures aerosolized droplets from the lower airways during normal breathing. It reflects the biochemical environment of the respiratory tract and provides access to lung-derived material without invasive procedures. Within this condensate, extracellular vesicles (EVs) constitute a particularly informative molecular source; protected by their lipid membrane, they preserve various biomolecules, enabling reliable multimarker analysis and sensitively reporting physiological and pathological changes.

Our research group profiles EBC-derived EVs and associated biomarkers to identify signatures of early disease diagnosis and progression. We use a standardized EBC collection protocol employing an FDA-approved device (RT Tube) with a sampling time of 10–15 minutes, optimized for EV yield and molecular integrity, ensuring samples suitable for clinical applications such as diagnosis and therapy monitoring.

To advance this work, we are establishing an EBC Biobank in collaboration with the University Cancer Center Hamburg, comprising samples from more than one hundred lung cancer patients collected before and after surgery, as well as healthy controls, including both smokers and non-smokers. This expanding resource provides a robust foundation for validating EV-based biomarkers and advancing EBC as a clinically meaningful tool for lung cancer diagnostics.

Building on this biobank, we apply complementary analytical approaches—from nucleic acid profiling to proteomic studies—to enable rigorous biomarker discovery and mechanistic insight. In parallel, we employ super-resolution imaging (dSTORM using the ONI Nanoimager) for single-EV characterization, allowing sensitive quantification of EV marker expression at high spatial resolution.