Summary of Study ST002179

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001387. The data can be accessed directly via it's Project DOI: 10.21228/M8FB0C This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST002179
Study Title	Impact of nitisinone on the cerebrospinal fluid metabolome of a murine model of alkaptonuria
Study Summary	Background: Nitisinone induced hypertyrosinaemia is well documented in Alkaptonuria (AKU), and there is uncertainty over whether it may contribute to a decline in cognitive function and or mood by altering neurotransmitter metabolism. The aim of this work was to evaluate the impact of nitisinone on the cerebrospinal fluid (CSF) metabolome in a murine model of AKU, with a view to providing additional insight into metabolic changes that occur following treatment with nitisinone. Methods: 17 CSF samples were collected from BALB/c Hgd-/-mice (n=8, treated with nitisinone – 4 mg/L and n=9, no treatment). Samples were diluted 1:1 with deionised water and analysed using a 1290 Infinity II liquid chromatography system coupled to a 6550 quadrupole time-of-flight mass spectrometry (Agilent, Cheadle, UK). Raw data were processed using a targeted feature extraction algorithm and an established in-house accurate mass retention time database. Matched entities (±10 ppm theoretical accurate mass and ±0.3 minutes retention time window) were filtered based on their frequency and variability. Experimental groups were compared using a moderated t-test with Benjamini-Hochberg false-discovery rate adjustment. Results: Tyrosine, acetyl-tyrosine, γ-glutamyl-tyrosine, p-hydroxyphenylacetic acid and 3-(4-hydroxyphenyl)lactic acid were shown to increase in abundance (log2 fold change 2.6-6.9, 3/5 were significant p<0.05) in the mice that received nitisinone. Several other metabolites of interest were matched but no significant differences were observed, including the aromatic amino acids phenylalanine and tryptophan, and monoamine metabolites adrenaline, 3-methoxy-4-hydroxyphenylglycol and octopamine. Conclusions: Evaluation of the CSF metabolome of a murine model of AKU showed a significant difference in the abundance of a limited number of metabolites. None of these have been reported in CSF from a murine model of AKU previously. Moreover this study confirms that some monoamine metabolites do not appear to be altered following nitisinone therapy.
Institute	University of Liverpool Institute of Life Course & Medical Sciences
Last Name	Davison
First Name	Andrew
Address	1. Department of Clinical Biochemistry and Metabolic Medicine, Liverpool Clinical Laboratories, Liverpool University Hospitals NHS Foundation Trust, Liverpool, UK; 2. Department of Musculoskeletal and Ageing Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool, UK 3. School of Exercise Science, Liverpool John Moores University, Liverpool, UK
Email	andrew.davison@liverpoolft.nhs.uk
Phone	0151 706 4011
Submit Date	2022-05-13
Num Groups	2
Total Subjects	17
Raw Data Available	Yes
Raw Data File Type(s)	d, mzML
Analysis Type Detail	LC-MS
Release Date	2022-06-08
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP002271
Sampleprep Summary:	Murine CSF samples were prepared by diluting 2 µL CSF with 2 µL deionized water (DIRECT-Q 3UV Millipore water purification system). CSF samples were pipetted directly into a 150 µL glass insert with polymer feet, which sat in a 2 mL amber screw vial (Agilent, Cheadle, UK). These vials were used to ensure maximal recovery of sample. Deionised water was also added directly to the glass insert, and mixed with the CSF sample using a pipette. The glass vials were centrifuged at 600 rpm for 10 min to ensure the diluted sample was at the bottom of the vial for sampling. 1 µL of diluted CSF was analysed in negative and positive polarities, respectively.

Sampleprep ID:

SP002271

Sampleprep Summary:

Murine CSF samples were prepared by diluting 2 µL CSF with 2 µL deionized water (DIRECT-Q 3UV Millipore water purification system). CSF samples were pipetted directly into a 150 µL glass insert with polymer feet, which sat in a 2 mL amber screw vial (Agilent, Cheadle, UK). These vials were used to ensure maximal recovery of sample. Deionised water was also added directly to the glass insert, and mixed with the CSF sample using a pipette. The glass vials were centrifuged at 600 rpm for 10 min to ensure the diluted sample was at the bottom of the vial for sampling. 1 µL of diluted CSF was analysed in negative and positive polarities, respectively.