Summary of Study ST004432

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 PR002800. The data can be accessed directly via it's Project DOI: 10.21228/M8QV7P This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php

Study ID	ST004432
Study Title	Metabolomic profiling of NMNAT2–SARM1 pathway modulation in mouse cortical neurodegeneration
Study 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 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 Name	Niou
First Name	Zhen-Xian
Address	400 EAST SEVENTH STREET
Email	niouz@iu.edu
Phone	7022097321
Submit Date	2025-11-06
Num Groups	5
Total Subjects	30
Num Males	30
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2026-01-02
Release Version	1

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Project:

Project ID:	PR002800
Project DOI:	doi: 10.21228/M8QV7P
Project Title:	Metabolomic profiling of NMNAT2–SARM1 pathway modulation in mouse cortical neurodegeneration
Project Summary:	Neuronal NAD⁺ depletion via NMNAT2 loss reprograms brain metabolism toward fatty-acid oxidation, lipid depletion, and oxidative stress. Multi-omics profiling (untargeted metabolomics and lipidomics) of NMNAT2 conditional knockout and SARM1-deleted mouse cortex revealed coordinated changes in glycolysis, redox cofactors, and lipid pathways. Complete SARM1 deletion restored metabolic and lipid balance, supporting the NMNAT2–SARM1 axis as a key regulator of neuronal and glial metabolism.
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:	SU004593
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	Treatment	Genotype
SA523867	IUB_ZN-07	Brain cortex	cKO	Nexcre/+ NMNAT2f/f
SA523868	IUB_ZN-10	Brain cortex	cKO	Nexcre/+ NMNAT2f/f
SA523869	IUB_ZN-09	Brain cortex	cKO	Nexcre/+ NMNAT2f/f
SA523870	IUB_ZN-08	Brain cortex	cKO	Nexcre/+ NMNAT2f/f
SA523871	IUB_ZN-06	Brain cortex	cKO	Nexcre/+ NMNAT2f/f
SA523856	IUB_ZN-22	Brain cortex	cKO S/+	Nexcre/+ NMNAT2f/f Sarm1null/+
SA523857	IUB_ZN-25	Brain cortex	cKO S/+	Nexcre/+ NMNAT2f/f Sarm1null/+
SA523858	IUB_ZN-24	Brain cortex	cKO S/+	Nexcre/+ NMNAT2f/f Sarm1null/+
SA523859	IUB_ZN-23	Brain cortex	cKO S/+	Nexcre/+ NMNAT2f/f Sarm1null/+
SA523860	IUB_ZN-21	Brain cortex	cKO S/+	Nexcre/+ NMNAT2f/f Sarm1null/+
SA523861	IUB_ZN-20	Brain cortex	cKO S/+	Nexcre/+ NMNAT2f/f Sarm1null/+
SA523862	IUB_ZN-26	Brain cortex	cKO S/S	Nexcre/+ NMNAT2f/f Sarm1null/null
SA523863	IUB_ZN-27	Brain cortex	cKO S/S	Nexcre/+ NMNAT2f/f Sarm1null/null
SA523864	IUB_ZN-28	Brain cortex	cKO S/S	Nexcre/+ NMNAT2f/f Sarm1null/null
SA523865	IUB_ZN-29	Brain cortex	cKO S/S	Nexcre/+ NMNAT2f/f Sarm1null/null
SA523866	IUB_ZN-30	Brain cortex	cKO S/S	Nexcre/+ NMNAT2f/f Sarm1null/null
SA523847	IUB_ZN-16	Brain cortex	Ctrl-2	Nexcre/+ NMNAT2f/+ Sarm1null/+
SA523848	IUB_ZN-18	Brain cortex	Ctrl-2	Nexcre/+ NMNAT2f/+ Sarm1null/+
SA523849	IUB_ZN-17	Brain cortex	Ctrl-2	Nexcre/+ NMNAT2f/+ Sarm1null/+
SA523850	IUB_ZN-19	Brain cortex	Ctrl-2	Nexcre/+ NMNAT2f/+ Sarm1null/+
SA523851	IUB_ZN-01	Brain cortex	Ctrl	Nexcre/+ NMNAT2f/+
SA523852	IUB_ZN-05	Brain cortex	Ctrl	Nexcre/+ NMNAT2f/+
SA523853	IUB_ZN-04	Brain cortex	Ctrl	Nexcre/+ NMNAT2f/+
SA523854	IUB_ZN-03	Brain cortex	Ctrl	Nexcre/+ NMNAT2f/+
SA523855	IUB_ZN-02	Brain cortex	Ctrl	Nexcre/+ NMNAT2f/+

Showing results 1 to 25 of 25

Showing results 1 to 25 of 25

Collection:

Collection ID:	CO004586
Collection Summary:	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:	TR004602
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:	SP004599
Sampleprep Summary:	Samples stored at -80°C was thawed on ice and homogenized in a ball-mill grinder at 30 Hz for 20s. 400 μL solution (Methanol : Water = 7:3, V/V) containing internal standard was mixed with 20 mg of ground sample and mixed in a shaker at 2500 rpm for 5 min. The mixture was placed on ice for 15 min and centrifuged at 12000 rpm for 10 min (4°C). 300 μL of the supernatant was collected and placed in -20°C for 30 min. The sample was then centrifuged at 12000 rpm for 3 min (4°C). A 200 μL aliquot of the supernatant was used for LC-MS

Sampleprep ID:

SP004599

Sampleprep Summary:

Samples stored at -80°C was thawed on ice and homogenized in a ball-mill grinder at 30 Hz for 20s. 400 μL solution (Methanol : Water = 7:3, V/V) containing internal standard was mixed with 20 mg of ground sample and mixed in a shaker at 2500 rpm for 5 min. The mixture was placed on ice for 15 min and centrifuged at 12000 rpm for 10 min (4°C). 300 μL of the supernatant was collected and placed in -20°C for 30 min. The sample was then centrifuged at 12000 rpm for 3 min (4°C). A 200 μL aliquot of the supernatant was used for LC-MS

Combined analysis:

Analysis ID	AN007413	AN007414
Chromatography ID	CH005617	CH005617
MS ID	MS007105	MS007106
Analysis type	MS	MS
Chromatography type	HILIC	HILIC
Chromatography system	Agilent 1290	Agilent 1290
Column	Waters ACQUITY UPLC BEH Amide (150 x 2.1 mm, 1.7 μm)	Waters ACQUITY UPLC BEH Amide (150 x 2.1 mm, 1.7 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:	CH005617
Instrument Name:	Agilent 1290
Column Name:	Waters ACQUITY UPLC BEH Amide (150 x 2.1 mm, 1.7 μm)
Column Temperature:	40°C
Flow Gradient:	0-3.5 min: 95% A, 5% B; 3.5-5.5 min: 70% A, 30% B; 5.5-6.5 min: 5% A, 95% B; 6.5-10 min: 95% A, 5% B.
Flow Rate:	0.40 mL/min
Solvent A:	60% acetonitrile/30% water/10% methanol; 20 mM ammonium formate; pH 10.6
Solvent B:	40% acetonitrile/60% water; 20 mM ammonium formate; pH 10.6
Chromatography Type:	HILIC

MS:

MS ID:	MS007105
Analysis ID:	AN007413
Instrument Name:	ABI Sciex Triple Quad 6500+
Instrument Type:	Triple TOF
MS Type:	ESI
MS Comments:	The original data file acquired by LC-MS was converted to mzML format by ProteoWizard. Peak extraction, peak alignment and retention time correction were performed by XCMS program. The peaks with missing rate >50% in each group of samples were filtered. The blank values were filled with KNN, and the peak area was corrected by SVR method. The metabolites were annotated by searching the MetwareBio’s in-house database, integrated public database, prediction database and metDNA. Finally, substances with a comprehensive identification score above 0.5 and a CV value of QC samples less than 0.3 were extracted, and then positive and negative mode were combined (substances with the highest qualitative grade and the lowest CV value were retained) to obtain the all_sample_data.xlsx file.
Ion Mode:	POSITIVE

MS ID:	MS007106
Analysis ID:	AN007414
Instrument Name:	ABI Sciex Triple Quad 6500+
Instrument Type:	Triple TOF
MS Type:	ESI
MS Comments:	The original data file acquired by LC-MS was converted to mzML format by ProteoWizard. Peak extraction, peak alignment and retention time correction were performed by XCMS program. The peaks with missing rate >50% in each group of samples were filtered. The blank values were filled with KNN, and the peak area was corrected by SVR method. The metabolites were annotated by searching the MetwareBio’s in-house database, integrated public database, prediction database and metDNA. Finally, substances with a comprehensive identification score above 0.5 and a CV value of QC samples less than 0.3 were extracted, and then positive and negative mode were combined (substances with the highest qualitative grade and the lowest CV value were retained) to obtain the all_sample_data.xlsx file.
Ion Mode:	NEGATIVE