Symposium Sessions
Day 3, June 12（Fri.） 15:50-16:10 Room B (4F 411+412)
- 3B-S2-1550
Metabolome alteration analysis in Niemann-Pick disease type C model cells using liquid chromatography/tandem mass spectrometry
(1Tohoku Univ., 2Tohoku Univ. Hosp.)
oMasahiro Watanabe1, Masamitsu Maekawa1,2, Nariyasu Mano1,2
Niemann-Pick disease type C (NPC) is a lysosomal lipid storage disorder caused by loss of NPC1/NPC2 function, resulting in lipid accumulation and dysfunction in multiple organs, including the central nervous system and liver. Although various lipid biomarkers have been identified, the global metabolic consequences of lipid accumulation remain unclear. Here, we profiled metabolite alterations in HepG2-based NPC1-knockout model cells using a stepwise workflow combining non-targeted metabolomics (NM) for discovery and targeted metabolomics (TM). NM highlighted candidate metabolites associated with NPC, and TM suggested alterations in multiple amino acids and related metabolites. Based on these TM findings, we further focused on tryptophan metabolism and developed a liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of tryptophan-derived metabolites encompassing the kynurenine and serotonin pathways. Optimization of LC conditions improved retention and separation, including isobaric separation. The method is being applied to quantify metabolites in NPC model cells and culture supernatants. Currently, proteomic analysis of pathway-related proteins is underway to integrate metabolite changes with enzyme expression and clarify the relationship between NPC pathology and tryptophan metabolic remodeling.