September 2019 - Gerald Stübiger

Dr. Gerald Stübiger

MedUni Wien RESEARCHER OF THE MONTH September 2019

MALDI-MS of extracellular vesicles enables differentiation of different degrees of chemoresistance in cancer cells.

In the current work matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS) was used for the analysis of protein profiles from extracellular vesicles (EVs) of cancer cells for the first time. EVs are cellular organelles in the nanometer range (~30-200 nm), which can be found in practically all human and animal body fluids (e.g. blood, urine, saliva, etc.) carrying diverse potential biomarker molecules (e.g. DNA, RNA, proteins, lipids, etc.).

As could be demonstrated using multivariate statistics   resistant and sensitive colon cancer cells as well as different levels of chemoresistance against 5-fluorouracil could be differentiated based upon the evaluation of protein patterns of the EVs. This suggests that EVs express specific proteins directly associated with adaptation of the cells to increasing doses of chemotherapy drugs, which may be associated with their proliferation in the body in the course of metastasis. Translated to a living organism, this constitutes a strong argument for the advantages of liquid biopsy (i.e. diagnostics from body fluids).

As the experiments showed, the quantities of EVs isolated from cell supernatants would be enough to measure a characteristic protein profile from a few millilitres of blood, using MALDI-MS. Due to the universal prevalence of EVs, there is a broad range of potential clinical and diagnostic applications for this technique which can be envisioned for the future. 

Selected Literature

1. Stübiger G., Nairn M. D., Abban T. K., Openshaw M. E., Mancera L., Herzig B., Wuczkowski M., Senfter D., Mader R. M. MALDI-MS Protein Profiling of Chemoresistance in Extracellular Vesicles of Cancer Cells. Anal. Chem. (2018), 90, 13178−13182. doi: 10.1021/acs.analchem.8b03756.
2. Fuh M., Heikaus L., Schlüter H. MALDI mass spectrometry in medical research and diagnostic routine laboratories. Int. J. Mass Spectrom. 2017, 416, 96-109.
3. Vlassov A. V., Magdaleno S., Setterquist R., Conrad R. Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochim. Biophys. Acta. 2012, 1820, 940-948.
4. Kreimer S., Belov A. M., Ghiran I., Murthy S. K., Frank D. A., Ivanov A. R. Mass-Spectrometry-Based Molecular Characterization of Extracellular Vesicles: Lipidomics and Proteomics. J. Proteome Res. 2015, 14, 2367−2384.
5. Schmidt W. M., Kalipciyan M., Dornstauder E., Rizovski B., Steger G. G., Sedivy R., Mueller M. W., Mader R.M. Dissecting progressive stages of 5-fluorouracil resistance in vitro using RNA expression profiling. Int. J. Cancer 2004, 112, 200-212.
6. Stübiger G., Marchetti M., Reichel C., Nagano M., Gmeiner G., Allmaier G. Molecular structural characterization of doping relevant recombinant human erythropoietins by means of matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry Rapid Commun. Mass  Spectrom. (2005) 19, 728–742.
7. Stübiger G., Belgacem O., Rehulka P., Bicker W., Binder B., Bochkov V. Analysis of Oxidized Phospholipids by MALDI Mass Spectrometry using 6-Aza-2-thiothymine together with Matrix Additives and disposable Target Surfaces. Anal. Chem. (2010) 82, 5502-5510.
8. Stübiger G., Wuczkowski M., Bicker W., Belgacem O. Nanoparticle-based Detection of Oxidized Phospholipids by MALDI mass spectrometry – Nano-MALDI approach. Anal. Chem. (2014), 86, 6401-6409.
9. Haller E., Stübiger G., Lafitte D., Lindner W., Lämmerhofer M. Chemical Recognition of Oxidation-Specific Epitopes in Low-Density Lipoproteins by a Nanoparticle Based Concept for Trapping, Enrichment, and Liquid Chromatography-Tandem Mass Spectrometry Analysis of Oxidative Stress Biomarkers. Anal Chem. (2014), 86, 9954-9961.
10. Stübiger G., Aldover-Macasaet E., Bicker W., Sobal G., Willfort-Ehringer A., Pock K., Bochkov V., Widhalm K., Belgacem B. Targeted profiling of atherogenic phospholipids in human plasma and lipoproteins of hyperlipidemic patients using MALDI-QIT-TOF-MS/MS. Atherosclerosis (2012) 224, 177-186.
11. Frey M.K., Alias S., Winter M. P., Redwan B., Stübiger G., Panzenboeck A., Alimohammadi A., Bonderman D., Jakowitsch J., Bergmeister H., Bochkov V., Preissner K. T., Lang I. M. Splenectomy Is Modifying the Vascular Remodeling of Thrombosis. J. Am. Heart Assoc. (2014), 3:e000772.
12. Buchmayer F., Schicker K., Steinkellner T., Geier P., Stübiger G., Hamilton P. J., Jurik A., Stockner T., Yang J. W., Montgomery T., Holy M., Hofmaier T., Kudlacek O., Matthies H. J., Ecker G. F., Bochkov V., Galli A., Boehm S., Sitte H. H. Amphetamine actions at the serotonin transporter rely on the availability of phosphatidylinositol-4,5-bisphosphate. Proc. Natl. Acad. Sci. U S A (2013), 110, 11642-11647.
13. Resch U., Tsatsaronis J. A., Le Rhun A., Stübiger G., Rohde M., Kasvandik S., Holzmeister S., Tinnefeld P., Wai S. N., Charpentier E. A Two-Component Regulatory System Impacts Extracellular Membrane-Derived Vesicle Production in Group A Streptococcus. mBio (2016), 7(6), e00207-16. doi: 10.1128/mBio.00207-16.
14. Stübiger G., Wuczkowski M., Mancera L., Lopandic K., Sterflinger K., Belgacem O. Characterization of yeasts and filamentous fungi using MALDI lipid phenotyping. J. Microbiol. Methods (2016), 130, 27-37.
15. Rossiter H., Stübiger G., Gröger M., König U., Gruber F., Sukseree S., Mlitz V., Buchberger M., Oskolkova O., Bochkov V., Eckhart L., Tschachler E. Inactivation of autophagy leads to changes in sebaceous gland morphology and function. Exp. Dermatol. (2018), 27(10):1142-1151. doi: 10.1111/exd.13752.
16. Wagner R., Stübiger G., Veigel D., Wuczkowski M., Lanzerstorfer P., Weghuber J., Karteris E., Nowikovsky K., Wilfinger N., Singer C. F., Colomer R., Benhamú B., López-Rodríguez M. L., Valent P., Grunt T. W. Multi-Level Silencing of Receptor-PI3K-mTORC1 by Fatty Acid Synthase Inhibitors is Crucial for their Efficacy against Ovarian Cancer Cells. Oncotarget (2017), 8:11600-11613. doi: 10.18632/oncotarget.14591.