Following on in silico and in vitro work, the effect of MCAD deficiency on CoA metabolism was investigated. Using a recently published HILIC-MS/MS method, free and acylated CoA species could be measured simultaneously in HepG2 MCAD-KO cells. The levels of CoA biosynthesis intermediates and total CoA was also characterised by HPLC in liver samples from MCAD-KO mice exposed to energetic stress (fasting adn cold). qPCR was applied to investigate changes in the CoA metabolism that might constitute reactions to an excessive degree of CoA sequestration. In silico modelling was used to hypothesise on the potential benefits of multifactorial, simultaneous adaptations as opposed to single-node adaptations.
DOI: None
Zenodo URL: None
Created at: 16th May 2025 at 11:45
Computational model (in silico)
No description specified
Model notebooks
Unzip model notebooks and keep in the same folder. Notebook "0, generate_model.nb" needs to be run before the others can be, as the model needs to be exported as Excel files before they cvan be used to make the figures.
- model_notebooks.zip
HepG2 cells (in vitro)
The behaviour of a published MCAD-KO HepG2 cell line was compared to that of a wild-type HepG2 cell line in terms of the changes in acyl-CoA and acylcarnitine levels (measured by HILIC-MS/MS) and gene expression pertaining to CoA metabolism (characterised by qPCR). Additionally, the incorporation of label from stable isotope-labelled pantothenate (vitamine B5) was over 24 hours of exposure to an energetic stressor was also investigated.
HepG2 acylcarnitine measurements, palmitate/no-glucose (Fig. 2, Fig. S2, Fig. S7)
No description specified
- data_Fig2_FigS2_FigS7_acylcarnitine_hepG2_palm_no_gluc.xlsx
HepG2 acyl-CoA measurements, palmitate/no-glucose (Fig. 2, Fig. S3, Fig. S7)
No description specified
- data_Fig2_FigS3_FigS7_acyl_CoA_hepG2_palm_no_gluc.xlsx
HepG2 CoA labelling data, palmitate/no-glucose (Fig. 3, Fig. S8, Tables S1-S4)
Incorporation of stable isotope from pantothenate into total and free CoA pools.
- data_Fig3_FigS8_TablesS1-S4_labelling_experiment_hepG2_palm_no_gluc.xlsx
HepG2 acyl-CoA measurements, palmitate/low-glucose (Fig.S5, Fig. S7)
No description specified
- data_FigS5_FigS7_acyl_CoA_hepG2_palm_low_gluc.xlsx
HepG2 acylcarnitine measurements, palmitate/low-glucose (Fig. S5, Fig. S7)
No description specified
- data_FigS5_FigS7_acylcarnitine_hepG2_palm_low_gluc.xlsx
HepG2 acyl-CoA/acylcarnitine measurements statistical significance, palmitate/low-glucose (Fig. S6)
No description specified
- data_FigS6_acyl_CoA_acylcarnitine_significance_hepG2_palm_low_gluc.xlsx
HepG2 acyl-CoA/acylcarnitine measurements statistical significance, palmitate/no-glucose (Fig. S4)
No description specified
- data_FigS4_acyl_CoA_acylcarnitine_significance_hepG2_palm_no_gluc.xlsx
HepG2 qPCR, palmitate/no-glucose (Fig. S9)
No description specified
- data_FigS9_hepG2_qPCR_palm_no_gluc.xlsx
HepG2 qPCR, palmitate/low-glucose (Fig. S11)
No description specified
- data_FigS11_hepG2_qPCR_palm_low_gluc.xlsx
HepG2 qPCR statistical significance, palmitate/no-glucose (Fig. S10)
No description specified
- data_FigS10_qPCR_significance_hepG2_palm_no_gluc.xlsx
HepG2 qPCR statistical significance, palmitate/low-glucose (Fig. S12)
No description specified
- data_FigS12_qPCR_significance_hepG2_palm_low_gluc.xlsx
Mice (in vivo)
MCAD-KO mice (C57BL/6J background) were exposed to three different conditions: "fed", "fasting", and "fasting + cold". In this study we determined the concentration of various CoA biosynthetic intermediates (including CoA) and the expression of CoA metabolic genes in liver tissue samples from these groups of mice.
Mice CoA biosynthetic intermediates (Fig. 4C)
No description specified
- data_Fig4C_mice_CoA_biosynthesis_intermediates.xlsx
Mice qPCR (Fig. 4E-F)
The expression of various genes related to the CoA metabolism as measured by qPCR.
- data_Fig4D-E_mice_qPCR.xlsx
These checksums allow you to check a Snapshot you have downloaded hasn't been modified. For details on how to use these please visit this guide
MD5: f6680b057ff86b6c8fff93532d8a8f6e
SHA1: f8279e208aa82e2f62ed86a807fd6c4003d8d0eb
Views: 61 Downloads: 6
Created: 16th May 2025 at 11:45