Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Stelzer, M.
Role of neutrophil extracellular trap formation in liver cirrhosis and sarcopenia
Humanmedizin; [ Diplomarbeit ] Medizinische Universität Graz; 2023. pp. 61
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Stadlbauer-Köllner Vanessa
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- Abstract:
- Background: Sarcopenia is a frequent complication of liver cirrhosis (LC), but it can also occur independently of any underlying cause. The immune system plays an important role in the pathogenesis of both LC and sarcopenia. Neutrophil extracellular traps (NETs) formation has been linked to chronic inflammation, and their function can be altered in people with LC; however, NETs formation in patients with sarcopenia has not been studied yet. Here, we aimed to study if NETs formation is altered in patients with sarcopenia and LC. Method: Neutrophil granulocytes from 121 patients were isolated and stimulated with heat-inactivated E. coli (250 bacteria/cell), phorbol 12-myristate 13-acetate (PMA) (100 nM), or incubation medium in duplicates for 2 hours at 37°C. Cells were fixed with paraformaldehyde and stained with 4′,6-diamidino-2-phenylindole (DAPI). Pictures of 10 random fields of vision per slide were taken with an Olympus BX51 Fluorescence Microscope at 600x total magnification. DNA area and NETosis analysis (DANA) algorithm was used to quantify the percentage of NETs formation per patient. Results: The DANA algorithm was successfully optimized and validated. The use of DANA decreased mean NETs formation analysis time from 30 minutes per patient (if conventional counting by eye is used) to 5 minutes (if DANA is used; p=0.006). There was no significant difference in NETs formation observed between the three groups of patients upon ex vivo stimulation of neutrophils with E. coli (p=0.778) or with PMA (p=0.490), but there was a tendency toward increased spontaneous NETs formation in the sarcopenia group (p=0.056). Conclusion: Artificial intelligence (AI) based approaches can be used for standardized and efficient analysis of NETs formation, making it feasible to introduce NETs formation analysis for large-scale clinical studies and as part of routine lab analysis in clinical practice. Spontaneous NETs formation within this study tended to be increased in patients with sarcopenia compared to patients with LC and LC + S. Though this finding did not reach statistical significance and has to be validated in further studies, it is the first evidence that NETs formation might be a potential biomarker of sarcopenia and might play a role in its pathogenesis.