Česká společnost pro hmotnostní spektrometrii

(CEEPC/IPM/CMSC 2022 - FrO-17)
Stability Study of Triacetylfusarinine C and Gliotoxin in bodily fluids

Jiří Houšť ^1,2, Tomáš Pluháček ^1,2, Anton Škríba ¹, Vladimír Havlíček ^1,2 *

1. Institute of Microbiology of the Czech Academy of Sciences, Prague, Czechia
2. Department of Analytical Chemistry, Faculty of Science, Palacký University, Olomouc, Czechia

Abstrakt

Introduction: During invasive pulmonary aspergillosis (IPA), Aspergillus fumigatus (Af) produces secondary metabolites supporting proliferation within a neutropenic host. Of note, extracellular siderophore triacetylfusarinine C (TafC) captures ferric cation [1], and mycotoxin gliotoxin (Gtx) fights host immunity [2]. Detection of these specific biomarkers in bodily fluids can improve early IPA diagnosis [3]. Methods: We tested the stability of TafC/Gtx in human urine (pH 6.44) and serum and rat urine (pH 7.86) at 37 °C in a one-week due course. We collected all data with Dionex UltiMate 3000 HPLC system coupled to a SolariX 12T FTICR and evaluated with DataAnalysis 5.0 and OriginPro 2021b. We calculated the TafC/Gtx half-life (t1/2) after data fitting with linear and exponential decay functions. Results: In human urine, TafC decay followed the zero-order kinetics with a t1/2 = 29.8 days. On the other hand, TafC decomposed 58.4x (t1/2 = 12.2 h) and 48.9x (t1/2 = 14.6 h) faster in the basic pH rat urine and human serum, respectively. The TafC degraded to triacetylfusarinine B and N2-acetyl-N5-anhydromevalonyl-N5-hydroxy-L-ornithine residues, which are simultaneously detected in bodily fluids for diagnostic purposes. Similarly, the most stable matrix for Gtx was human urine (t1/2 = 5.8 days) compared to rat urine and human serum, where 80 and 94% of Gtx decomposed within one day into unknown metabolites, respectively. Conclusion: A slightly acidic human urine represents a suitable matrix for preserving the clinically valuable Af virulence factors secreted during proliferation in a host. The simultaneous detection of secreted secondary metabolites and their degradation products in urine could play an essential role in IPA diagnostics even in the early stages of infection.

* Korespondující autor: jiri.houst@biomed.cas.cz

Literatura

1. Misslinger M. et al.: Biochim Biophys Acta Mol Cell Res 1868(1), 118885 (2021).
2. Sugui J.A. et al.: Sci Rep 7(1), 10977 (2017).
3. Patil R.H. et al.: Mass Spectrom Rev (2021). DOI: 10.1002/mas.21755

Poděkování:

We acknowledge the support from Czech Science Foundation, grant No. 21-17044S.