Summary of Study ST004451

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 PR002811. The data can be accessed directly via it's Project DOI: 10.21228/M89P0K This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php

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Study IDST004451
Study TitleLipidomics profiling of NMNAT2–SARM1 pathway modulation in mouse cortical neurodegeneration
Study SummaryNAD⁺ homeostasis is vital for neuronal health, as demonstrated by the opposing roles of nicotinamide mononucleotide adenylyltransferase 2 (NMNAT2), a NAD⁺-synthesizing enzyme, and sterile alpha and TIR motif-containing protein 1 (SARM1), a NAD⁺ hydrolase. Neurodegenerative insults that decrease NMNAT2 activate SARM1, leading to axonal degeneration. To determine how the NMNAT2–SARM1 axis influences brain energy and lipid metabolism, we performed untargeted metabolomics and targeted lipidomics on cerebral cortices collected from postnatal day 16-21 (P16-P21) mice. Experimental groups included control (Nex-Cre; Nmnat2f/+), NMNAT2 conditional knockout (Nex-Cre; Nmnat2f/f), and Sarm1 heterozygous or homozygous deletions in the NMNAT2 cKO background. Each condition included 4–6 biological replicates, with samples processed in parallel under identical growth and housing conditions. Loss of NMNAT2 in glutamatergic neurons resulted in a brain-wide metabolic shift from glucose to lipid catabolism, reductions in key glycolytic intermediates, and widespread depletion of cholesterol-, sphingolipid-, and phospholipid-related species. Metabolomic profiling revealed decreased NAD⁺, impaired glycolysis, and accumulation of fatty-acid oxidation and ketone metabolites. These metabolic disturbances were accompanied by altered glial expression of lipid and glucose metabolism genes, increased inflammatory signaling, and disrupted astrocytic transcriptomic profiles related to cholesterol biosynthesis and immune activation. Notably, complete SARM1 deletion normalized cortical NAD⁺ levels, restored lipid metabolic profiles, reduced ketone and FAO intermediates, and attenuated astrocytic inflammatory signatures, leading to improved axonal integrity and motor function. Together, these results demonstrate that neuronal NAD⁺ depletion triggers maladaptive, SARM1-dependent metabolic reprogramming that shifts energy use from glucose to lipids, thereby promoting inflammation and neurodegeneration.
Institute
Indiana University Bloomington
Last NameNiou
First NameZhen-Xian
Address400 EAST SEVENTH STREET, BLOOMINGTON, IN, 47405, USA.
Emailniouz@iu.edu
Phone7022097321
Submit Date2025-11-24
Num Groups5
Total Subjects30
Num Males30
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2026-01-06
Release Version1
Zhen-Xian Niou Zhen-Xian Niou
https://dx.doi.org/10.21228/M89P0K
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002811
Project DOI:doi: 10.21228/M89P0K
Project Title:Lipidomics profiling of NMNAT2–SARM1 pathway modulation in mouse cortical neurodegeneration
Project Summary:NAD⁺ homeostasis is vital for neuronal health, as demonstrated by the opposing roles of nicotinamide mononucleotide adenylyltransferase 2 (NMNAT2), a NAD⁺-synthesizing enzyme, and sterile alpha and TIR motif-containing protein 1 (SARM1), a NAD⁺ hydrolase. Neurodegenerative insults that decrease NMNAT2 activate SARM1, leading to axon loss. To understand how the NMNAT2–SARM1 axis influences brain energy metabolism, we employed multi-omics approaches to investigate the metabolic changes resulting from neuronal NMNAT2 loss. Loss of NMNAT2 in glutamatergic neurons leads to a significant metabolic shift in the cerebral cortex from glucose to lipid catabolism, reduced lipid abundance, and pronounced neurodegenerative phenotypes and motor behavioral deficits. These metabolic disturbances are accompanied by altered glial expression of enzymes regulating glucose and lipid metabolism, enhanced inflammatory signaling, and disrupted astrocytic transcriptomic profiles related to cholesterol synthesis and immune activation. Notably, SARM1 deletion in NMNAT2-deficient mice restored lipid metabolism, astrocyte transcriptomic profiles, and mitigated neurodegeneration and motor behaviors. These findings suggest that neuronal NAD⁺ depletion triggers maladaptive, SARM1-dependent metabolic reprogramming, shifting energy use from glucose to lipids, which in turn promotes inflammation and neurodegeneration. Metabolomic profiling revealed decreased NAD⁺ and glycolytic intermediates, accumulation of fatty acid oxidation and ketone metabolites, and widespread depletion of cholesterol- and phospholipid-related species, all of which were normalized by SARM1 deletion.
Institute:Indiana University Bloomington
Last Name:Niou
First Name:Zhen-Xian
Address:400 EAST SEVENTH STREET, BLOOMINGTON, IN, 47405, USA.
Email:niouz@iu.edu
Phone:7022097321

Subject:

Subject ID:SU004613
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Genotype Treatment
SA526943cKOB-1Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/null cKO S/S
SA526944cKOB-2Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/null cKO S/S
SA526945cKOB-3Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/null cKO S/S
SA526946cKOB-4Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/null cKO S/S
SA526947cKOB-5Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/null cKO S/S
SA526937cKOA-3Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/+ cKO S/+
SA526938cKOA-6Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/+ cKO S/+
SA526939cKOA-5Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/+ cKO S/+
SA526940cKOA-4Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/+ cKO S/+
SA526941cKOA-2Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/+ cKO S/+
SA526942cKOA-1Brain cortex Nexcre/+ NMNAT2f/f Sarm1null/+ cKO S/+
SA526948cKO-2Brain cortex Nexcre/+ NMNAT2f/f cKO
SA526949cKO-5Brain cortex Nexcre/+ NMNAT2f/f cKO
SA526950cKO-4Brain cortex Nexcre/+ NMNAT2f/f cKO
SA526951cKO-3Brain cortex Nexcre/+ NMNAT2f/f cKO
SA526952cKO-1Brain cortex Nexcre/+ NMNAT2f/f cKO
SA526928CtrlA-1Brain cortex Nexcre/+ NMNAT2f/+ Sarm1null/+ Ctrl-3
SA526929CtrlA-3Brain cortex Nexcre/+ NMNAT2f/+ Sarm1null/+ Ctrl-3
SA526930CtrlA-2Brain cortex Nexcre/+ NMNAT2f/+ Sarm1null/+ Ctrl-3
SA526931CtrlA-4Brain cortex Nexcre/+ NMNAT2f/+ Sarm1null/+ Ctrl-3
SA526932Ctrl-1Brain cortex Nexcre/+ NMNAT2f/+ Ctrl
SA526933Ctrl-5Brain cortex Nexcre/+ NMNAT2f/+ Ctrl
SA526934Ctrl-4Brain cortex Nexcre/+ NMNAT2f/+ Ctrl
SA526935Ctrl-3Brain cortex Nexcre/+ NMNAT2f/+ Ctrl
SA526936Ctrl-2Brain cortex Nexcre/+ NMNAT2f/+ Ctrl
Showing results 1 to 25 of 25

Collection:

Collection ID:CO004606
Collection Summary:Animals were genotyped using ear punch tissue followed by PCR-based genotyping. All the Brain tissues were collected when mice reached postnatal day 16–21. The somatosensory cortical region was rapidly dissected immediately after euthanasia. All procedures were performed on ice to preserve tissue integrity. The dissected tissues were then snap-frozen in a dry ice/ethanol bath and stored at -80°C.
Sample Type:Brain cortex

Treatment:

Treatment ID:TR004622
Treatment Summary:No additional treatment conducted, samples are clustered by genotype. Animals were genotyped using ear punch tissue followed by PCR-based genotyping.

Sample Preparation:

Sampleprep ID:SP004619
Sampleprep Summary:Samples stored at -80°C was thawed on ice. Multi-point sample and weigh 20 mg of sample, homogenize (30 HZ) for 20 s with a steel ball and the centrifuge (3000 rpm, 4°C) for 30 s. Then add 1 mL of the extraction solvent (MTBE: MeOH =3:1, v/v) containing internal standard mixture. After whirling the mixture for 15 min, 200 μL of ultrapure water was added. Vortex for 1 min and centrifuge at 12,000 rpm for 10 min. 200 μL of the upper organic layer was collected and evaporated using a vacuum concentrator. The dry extract was dissolved in 200 μL reconstituted solution (ACN: IPA=1:1, v/v) to LC-MS/MS analysis.

Combined analysis:

Analysis ID AN007454 AN007455
Chromatography ID CH005645 CH005645
MS ID MS007146 MS007147
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Agilent 1290 Agilent 1290
Column Thermo Accucore HILIC (100 x 2.1 mm, 2.6 μm) Thermo Accucore HILIC (100 x 2.1 mm, 2.6 μm)
MS Type ESI ESI
MS instrument type Triple TOF Triple TOF
MS instrument name ABI Sciex Triple Quad 6500+ ABI Sciex Triple Quad 6500+
Ion Mode POSITIVE NEGATIVE
Units Peak area (arbitrary units) Peak area (arbitrary units)

Chromatography:

Chromatography ID:CH005645
Instrument Name:Agilent 1290
Column Name:Thermo Accucore HILIC (100 x 2.1 mm, 2.6 μm)
Column Temperature:45°C
Flow Gradient:80% A, 20% B at 0 min; 70% A, 30% B at 2 min; 40% A, 60% B at 4 min; 15% A, 85% B at 9 min; 10% A, 90% B at 14 min; 5% A, 95% B at 15.5 min; 5% A, 95% B at 17.3 min; 80% A, 20% B at 17.5 min; 80% A, 20% B at 20 min.
Flow Rate:0.35 mL/min
Solvent A:60% Acetonitrile/40% Water; 0.1% formic acid; 10 mmol/L ammonium formate
Solvent B:90% Isopropyl alcohol/10% Acetonitrile; 0.1% formic acid; 10 mmol/L ammonium formate
Chromatography Type:HILIC

MS:

MS ID:MS007146
Analysis ID:AN007454
Instrument Name:ABI Sciex Triple Quad 6500+
Instrument Type:Triple TOF
MS Type:ESI
MS Comments:LIT and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (QTRAP), QTRAP® 6500+ LC-MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operating in positive and negative ion mode and controlled by Analyst 1.6.3 software (Sciex). The ESI source operation parameters were as follows: ion source, turbo spray; source temperature 500°C; ion spray voltage (IS) 5500 V(Positive),-4500 V(Negative); Ion source gas 1 (GS1), gas 2 (GS2), curtain gas (CUR) were set at 45, 55, and 35 psi, respectively. Instrument tuning and mass calibration were performed with 10 and 100 μmol/L polypropylene glycol solutions in QQQ and LIT modes, respectively. QQQ scans were acquired as MRM experiments with collision gas (nitrogen) set to 5 psi. DP and CE for individual MRM transitions was done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the lipids eluted within this period.
Ion Mode:POSITIVE
  
MS ID:MS007147
Analysis ID:AN007455
Instrument Name:ABI Sciex Triple Quad 6500+
Instrument Type:Triple TOF
MS Type:ESI
MS Comments:LIT and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (QTRAP), QTRAP® 6500+ LC-MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operating in positive and negative ion mode and controlled by Analyst 1.6.3 software (Sciex). The ESI source operation parameters were as follows: ion source, turbo spray; source temperature 500°C; ion spray voltage (IS) 5500 V(Positive),-4500 V(Negative); Ion source gas 1 (GS1), gas 2 (GS2), curtain gas (CUR) were set at 45, 55, and 35 psi, respectively. Instrument tuning and mass calibration were performed with 10 and 100 μmol/L polypropylene glycol solutions in QQQ and LIT modes, respectively. QQQ scans were acquired as MRM experiments with collision gas (nitrogen) set to 5 psi. DP and CE for individual MRM transitions was done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the lipids eluted within this period.
Ion Mode:NEGATIVE
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