Wystąpienie ustne Magdalena Elżbieta Marzec (BIO-Wt)
TOF-SIMS TESTING OF THE MOLECULAR COMPOSITION OF URINARY EXTRACELLULAR VESICLES TO DETERMINE SYMPTOMS OF DIABETIC KIDNEY DISEASE
1 Department of Medical Physics, Institute of Physics M. Smoluchowskiego, Jagiellonian University, Łojasiewicza, 30-348 Krakow, Polska
2 Department of Metabolic Diseases, Jagiellonian University Medical College, Jakubowskiego 2, 30-688 Krakow, Polska
We analyzed extracellular vesicles (EVs) isolated from the morning urine of patients with type 1 diabetes mellitus (T1DM). EVs are spherical cellular structures surrounded by a lipid bilayer with sizes from 30 to 1000 nm. Due to the constantly growing importance of EVs in the diagnosis and treatment of diseases, more attempts are made to effectively isolate, detect and analyze of EVs . Chronic kidney disease (CKD) is currently an increasing clinical problem. Hence, there is a need to identify the early biomarkers of CKD. EVs can serve as non-invasive indicators to detect changes in a kidney and implement effective treatment before irreversible structural changes in the kidney occur. However, the preparation of EVs for nanoscale research is difficult for several reasons. First, the isolation of EVs from the conditioned medium where the concentration of EVs is low. Here, EVs were purified by a Low-Vacuum Filtration (L-VF) technique and concentrated by the ultracentrifugation process . Second, it is the isolation of individual molecular components, like lipids, which are sensitive to each stage of preparation. Therefore, the possibility of testing them in their native state, without the need for extraction, is extremely promising. In the case of Time of Flight - Secondary Ion Mass Spectrometry (ToF-SIMS), there is no need to perform such preparations or labelling, which allows the structure to be examined in a semi-native state .
The aim of the study is to develop a method of early detection of the first microvascular symptoms including CKD. The detection procedure will be based on the use of two physical methods, Raman Spectroscopy (SR) and ToF-SIMS to precisely determine the molecular compounds whose EVs composition changes during early kidney damage. The study recruited 36 patients with T1DM at the University Hospital in Krakow (no. 1072.6120.109.2019). Here we present the preliminary results of ToF-SIMS analysis of urinary EVs, showing the diversity in individual amino acids and the lipid content.
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 A. Kamińska, Molecular characteristics of platelet and urinary extracellular vesicles and their possible applications in nanomedicine, 2019, PhD thesis, Jagiellonian University, Krakow.
 M.E. Marzec, D. Wojtysiak, K. Połtowicz, J. Nowak, ToF-SIMS spectrometry to observe fatty acid profiles of breast tissues in broiler chicken subjected to varied vegetable oil diet, J. Mass Spectrom., 55 (2020).
This work was supported by the SciMat PRA MNS mini-grant the qLife grant funded under the Strategic Programme Excellence Initiative at UJ and from the CMUJ statutory funds (no. N41/DBS/000068).