여름정기학술대회
2022여름초록
발표자 및 발표 내용
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공동저자
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접수자
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The
endogenous amino acids (AAs) level can reflect biological metabolism and
function including diseases and damage. A large number of systemically
administered drugs have the potential to cause drug-induced lung injury, and
the AAs level can be expected as effective biomarkers of toxicity in organs
including lung. Lung organoids provide an excellent platform to model injury to
and repair of lungs. This study investigates and porposes the endogenous AAs
level as biomarkers for toxicity of lung using organoids model. For AA analysis,
we describe the comprehensive profiling methods of seventeen AAs (proline, valine,
leucine, phenylalanine, isoleucine, tyrosine, histidine, methionine, aspartic
acid, threonine, arginine, tryptophan, glutamic acid, glutamine, alanine, glycine,
and serine) based on the isotope dilution liquid chromatography-mass
spectrometry (ID-LC-MS). The two methods for both derivatized AAs and non-derivatized
AAs were validated and verified with certified reference materials, followed by
analysis of lung organoids and the cell media. The samples for both methods
were treated by protein precipitation reagents, 15 % v/v 5-sulfosalycylic acid,
or 75 % v/v acetonitrile. All target AA residues were
clearly separated within 10 min in both LC-MS conditions. The lung
organoids were treated with drugs (Bleomycin 2 µg/mL for 48 h, and Bleomycin 5 µg/mL
for 72 h) for inducing lung injury, and the AA contents in both cell lysate and
media were quantified and normalized by protein concentration. The AA levels
were compared between drug treated group and control group, and some residues
were classified as candidate biomarkers of toxicity monitoring. The reliability
of the target residues will be confirmed with repeated trials, and it will be
proposed as a new marker for toxicity monitoring in lung organoids.
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