Summary of Study ST000792

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 PR000575. The data can be accessed directly via it's Project DOI: 10.21228/M8BM2Z 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.

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Study IDST000792
Study TitleLarge Untargeted Profiling of Myelin to Enhance Recovery of Function after SCI
Study SummaryTissue is from adult mouse spinal cord (SC). We are submitting these samples for Untargeted Profiling (unbiased metabolomics assay) and for lipid analysis. The lipid assays we request are 1) free fatty acid composition of lipids; 2) free fatty acid panel; 3) cholesterol concentration (free and bound); 4) Ceramides, including galactosyl and glucosyl; 5) sphingomyelin. The Untargeted profiling is our top priority, followed by the lipid assays as listed. All samples were snap frozen at the point of harvest and approximate weights are provided. The samples are submitted as intact pieces of tissue. There are 20 samples total, n=5 for each group that includes LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet).
Institute
Mayo Clinic
Last NameScarisbrick
First NameIsobel
Address200 First St. SW, Rochester, Minnesota, 55905, USA
Emailscarisbrick.isobel@mayo.edu
Phone507-284-0124
Submit Date2017-07-11
Analysis Type DetailLC-MS
Release Date2021-01-19
Release Version1
Isobel Scarisbrick Isobel Scarisbrick
https://dx.doi.org/10.21228/M8BM2Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR000575
Project DOI:doi: 10.21228/M8BM2Z
Project Title:Mayo Pilot and Feasibility: Targeting Myelin Metabolism to Enhance Recovery of Function after SCI
Project Summary:The loss of myelin, degeneration of the myelin producing oligodendroglia and impaired remyelination are essential features of traumatic spinal cord injury (SCI) that significantly limit patient recovery of function. The lipid rich composition of myelin, including exceptionally high levels of saturated fatty acids, underlie its essential physiological roles, including its structural and signaling properties and electrical insulation of axons to facilitate the conduction of nerve impulses. The myelin sheaths also provide metabolic support to the axons they wrap, and myelin health is therefore essential to the maintenance of axon integrity and function in the brain and spinal cord. The primary goal of this Pilot Proposal to the Mayo Clinic Metabolomics Core is to integrate highly sensitive metabolomics liquid chromatography-tandem mass spectrometry (LC/MS/MS) assays to quantify the lipid composition of the myelin membrane, with our conventional neurobehavioral approaches, enabling us to explore the metabolic basis of new interventions capable of promoting myelin regeneration and restoration of patient function. Metabolomics Core expertise in Magnetic Resonance Spectroscopy (NMR) based evaluation of key metabolites involved in CNS injury and repair (N-acetyl-L-aspartate, choline, myo-inositol, glucose/ glutamine and lactate) will also be applied to strengthen our mechanistic understanding of myelin injury and repair. Specifically, utilizing these innovative approaches we will test a novel hypothesis driven by new preliminary findings that the levels of dietary fatty acids can be optimized alone, or in combination with exercise training, to facilitate myelin regeneration and recovery of neurobehavioral function after injury to the adult spinal cord. In Aim 1, we will determine whether alterations in dietary fat, including saturated fat or omega-3 fatty acids, facilitate restoration of the myelin membrane and metabolite signatures of central nervous system repair after experimental SCI in adult mice. In Aim 2, we will determine whether exercise training alone or in combination with dietary fatty acid supplementation fosters myelin regeneration and recovery of function after experimental SCI. The proposed studies will leverage the expertise of the Mayo Metabolomics Core with that of Dr. Scarisbrick (Mayo) in myelin biology and Dr. Gomez Pinilla (UCLA) in central nervous system plasticity to investigate whether two highly targetable lifestyle variables, that is diet and exercise, can be modulated to improve myelin metabolism and functional recovery after SCI.
Institute:Mayo Clinic
Last Name:Scarisbrick
First Name:Isobel
Address:200 First St. SW, Rochester, Minnesota, 55905, USA
Email:scarisbrick.isobel@mayo.edu
Phone:507-284-0124

Subject:

Subject ID:SU000817
Subject Type:Mouse
Subject Species:Mus musculus
Taxonomy ID:10090
Species Group:Mammal

Factors:

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

mb_sample_id local_sample_id Group rep
SA043617nC18-2jun16-012-r001HFD 1
SA043618philic-31may16-007-r001HFD 1
SA043619nhilic-1jun16-007-r001_1HFD 1
SA043620nhilic-1jun16-012-r001_1HFD 1
SA043621nhilic-1jun16-014-r001_1HFD 1
SA043622nC18-2jun16-016-r001HFD 1
SA043623nhilic-1jun16-016-r001_1HFD 1
SA043624philic-31may16-012-r001HFD 1
SA043625nC18-2jun16-014-r001HFD 1
SA043626pC18-31may16-016-r001HFD 1
SA043627philic-31may16-016-r001HFD 1
SA043628nC18-2jun16-007-r001HFD 1
SA043629philic-31may16-014-r001HFD 1
SA043630pC18-31may16-007-r001HFD 1
SA043631pC18-31may16-014-r001HFD 1
SA043632pC18-31may16-012-r001HFD 1
SA043633nhilic-1jun16-014-r002_1HFD 2
SA043634pC18-31may16-012-r002HFD 2
SA043635nhilic-1jun16-007-r002_1HFD 2
SA043636nC18-2jun16-016-r002HFD 2
SA043637pC18-31may16-016-r002HFD 2
SA043638pC18-31may16-007-r002HFD 2
SA043639pC18-31may16-014-r002HFD 2
SA043640nhilic-1jun16-012-r002_1HFD 2
SA043641nhilic-1jun16-016-r002_1HFD 2
SA043642nC18-2jun16-012-r002HFD 2
SA043643philic-31may16-007-r002HFD 2
SA043644philic-31may16-012-r002HFD 2
SA043645nC18-2jun16-007-r002HFD 2
SA043646philic-31may16-014-r002HFD 2
SA043647nC18-2jun16-014-r002HFD 2
SA043648philic-31may16-016-r002HFD 2
SA043649pC18-31may16-013-r001HFHS 1
SA043650philic-31may16-013-r001HFHS 1
SA043651philic-31may16-019-r001HFHS 1
SA043652philic-31may16-010-r001HFHS 1
SA043653nC18-2jun16-004-r001HFHS 1
SA043654philic-31may16-004-r001HFHS 1
SA043655nhilic-1jun16-019-r001_1HFHS 1
SA043656nhilic-1jun16-013-r001_1HFHS 1
SA043657nhilic-1jun16-010-r001_1HFHS 1
SA043658nhilic-1jun16-008-r001_1HFHS 1
SA043659pC18-31may16-019-r001HFHS 1
SA043660pC18-31may16-010-r001HFHS 1
SA043661philic-31may16-008-r001HFHS 1
SA043662nhilic-1jun16-004-r001_1HFHS 1
SA043663nC18-2jun16-008-r001HFHS 1
SA043664pC18-31may16-008-r001HFHS 1
SA043665pC18-31may16-004-r001HFHS 1
SA043666nC18-2jun16-010-r001HFHS 1
SA043667nC18-2jun16-019-r001HFHS 1
SA043668nC18-2jun16-013-r001HFHS 1
SA043669nhilic-1jun16-019-r002_1HFHS 2
SA043670nhilic-1jun16-004-r002_1HFHS 2
SA043671philic-31may16-010-r002HFHS 2
SA043672nhilic-1jun16-008-r002_1HFHS 2
SA043673nhilic-1jun16-010-r002_1HFHS 2
SA043674nhilic-1jun16-013-r002_1HFHS 2
SA043675philic-31may16-004-r002HFHS 2
SA043676pC18-31may16-010-r002HFHS 2
SA043677nC18-2jun16-010-r002HFHS 2
SA043678nC18-2jun16-019-r002HFHS 2
SA043679nC18-2jun16-008-r002HFHS 2
SA043680philic-31may16-008-r002HFHS 2
SA043681pC18-31may16-004-r002HFHS 2
SA043682pC18-31may16-013-r002HFHS 2
SA043683pC18-31may16-019-r002HFHS 2
SA043684pC18-31may16-008-r002HFHS 2
SA043685philic-31may16-019-r002HFHS 2
SA043686nC18-2jun16-004-r002HFHS 2
SA043687philic-31may16-013-r002HFHS 2
SA043688nC18-2jun16-013-r002HFHS 2
SA043689pC18-31may16-009-r001Keto 1
SA043690pC18-31may16-005-r001Keto 1
SA043691nhilic-1jun16-005-r001_1Keto 1
SA043692philic-31may16-005-r001Keto 1
SA043693nC18-2jun16-018-r001Keto 1
SA043694nhilic-1jun16-015-r001_1Keto 1
SA043695nC18-2jun16-005-r001Keto 1
SA043696pC18-31may16-018-r001Keto 1
SA043697pC18-31may16-015-r001Keto 1
SA043698nC18-2jun16-015-r001Keto 1
SA043699philic-31may16-015-r001Keto 1
SA043700philic-31may16-009-r001Keto 1
SA043701nhilic-1jun16-018-r001_1Keto 1
SA043702philic-31may16-018-r001Keto 1
SA043703nhilic-1jun16-009-r001_1Keto 1
SA043704nC18-2jun16-009-r001Keto 1
SA043705nhilic-1jun16-018-r002_1Keto 2
SA043706nC18-2jun16-009-r002Keto 2
SA043707philic-31may16-018-r002Keto 2
SA043708nC18-2jun16-005-r002Keto 2
SA043709philic-31may16-015-r002Keto 2
SA043710philic-31may16-009-r002Keto 2
SA043711philic-31may16-005-r002Keto 2
SA043712nhilic-1jun16-015-r002_1Keto 2
SA043713pC18-31may16-005-r002Keto 2
SA043714pC18-31may16-018-r002Keto 2
SA043715nhilic-1jun16-009-r002_1Keto 2
SA043716nhilic-1jun16-005-r002_1Keto 2
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Collection:

Collection ID:CO000811
Collection Summary:Uninjured or SCI mice will be randomly assigned to one of four groups, LF (low fat diet); HF (high fat diet); HFHS (high fat high sucrose diet); and Keto (ketogenic diet). All diets will be obtained from Research Diets, NJ USA43,45. Dietary fat supplementation will be initiated at 1 week after SCI. This time point for intervention was chosen to provide a meaningful timeframe for clinical translation. Also, a 1 week period will allow time for mice to recover prior to providing access to wheel running (Aim 2). The impact of dietary fat supplementation on myelin metabolism will be examined after a period of 7 weeks, including determination of (i) the lipid profile of the myelin membrane using LC/MS/MS; and (ii) metabolic markers of spinal cord metabolism by NMR. Results will be correlated with (iii) cellular and molecular markers of spinal cord pathophysiology including the appearance of OPCs, oligodendroglia and myelin, axon health, astrogliosis and inflammation; and (iv) the extent of sensorimotor recovery. (v) In addition, to gauge the impact of the dietary fat on systemic metabolic status, Insulin and Glucose Resistance Tests will be performed at 7 weeks. Food intake (g/day) and body weight gain (% initial weight) will be measured daily until the endpoint of each experiment. Mice will be housed individually in a temperature-controlled facility with a 12:12-h light-dark cycle and ad libitum access to each diet and water. A Power Analysis was performed based on histological outcomes in mice with contusion compression injury. To detect a difference between groups of 20%, which would very meaningful, a group size of 8 will be needed to achieve a power of 0.85. An additional 2 mice per group has been added to account for mortality. The Mayo Clinic Institutional Animal Care and Use Committee has approved of the proposed studies.
Sample Type:Spinal cord

Treatment:

Treatment ID:TR000831
Treatment Summary:To test the hypothesis that optimizing dietary fat will facilitate myelin repair after SCI, the diet of uninjured adult female C57BL6/J mice (12 week, 22-25g, Jackson), or those with experimental contusion-compression SCI of the lumbosacral spinal cord (L2-L3) (Fejota Clip 3g Force, applied for 30s)32,47 will be supplemented with saturated fat. The 3g Clip produces moderate SCI including demyelination and clinical impairment and we recently published a detailed methodology. At 1 week after injury, the 3g injured mice are expected to have an average Basso Mouse Scale score (BMS)=5 on a 9 point scale such that they have frequent plantar stepping with no or some coordination. This level of impairment was chosen to provide a sufficient window to observe recovery, and to be at a level compatible with examination of exercise training by wheel running (Aim 2).

Sample Preparation:

Sampleprep ID:SP000824
Sampleprep Summary:large scale profiling of mouse spinal cord Lipids will be quantified in myelin isolated in high yield and purity by subcellular fractionation from the lumbosacral spinal cord. While there are no absolutely ‘myelin-specific’ lipids, galactocerebroside is the most typical of myelin in the adult nervous system being directly proportional to the amount of myelin. Sulfatide is another galactolipid enriched in myelin. Together with cholesterol, these form 78% of the total amount of lipid in the myelin membrane and each will be quantified using LC/MS/MS. A highly sensitive assay for galactocerebroside was recently established by the Mayo Metabolomics Core and can be implemented immediately. The LC/MS/MS panel for free fatty acids, including the very long chain fatty acids found in myelin is also routinely performed by the Core. Cholesterol will be quantified using an NMR-based approach by the Mayo Dept. of Laboratory Medicine Clinical Core. Additionally, we have a plan in place with the Metabolomics Core to develop LC/MS/MS assays for sulfatide and sphingomyelin during the Pilot proposal. Having quantitative assays for each of these key myelin lipids will facilitate our goal to comprehensively profile myelin lipid metabolism and will form foundational assays for a future NIH grant focused on myelin metabolism.

Combined analysis:

Analysis ID AN001260 AN001261 AN001262 AN001263
Analysis type MS MS MS MS
Chromatography type Reversed phase Reversed phase HILIC HILIC
Chromatography system Agilent 1290 Infinity Agilent 1290 Infinity Agilent 1290 Infinity Agilent 1290 Infinity
Column Waters Acquity BEH Amide (150 x 2.1mm,1.7um) Waters Acquity BEH Amide (150 x 2.1mm,1.7um) Waters Acquity HSS C18 (150 x 2.1mm,1.8um) Waters Acquity HSS C18 (150 x 2.1mm,1.8um)
MS Type ESI ESI ESI ESI
MS instrument type QTOF QTOF QTOF QTOF
MS instrument name Agilent 6550 QTOF Agilent 6550 QTOF Agilent 6550 QTOF Agilent 6550 QTOF
Ion Mode POSITIVE NEGATIVE POSITIVE NEGATIVE
Units intensity intensity intensity intensity

Chromatography:

Chromatography ID:CH000878
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
Chromatography Type:Reversed phase
  
Chromatography ID:CH000879
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity HSS C18 (150 x 2.1mm,1.8um)
Chromatography Type:HILIC

MS:

MS ID:MS001153
Analysis ID:AN001260
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS001154
Analysis ID:AN001261
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
  
MS ID:MS001155
Analysis ID:AN001262
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS001156
Analysis ID:AN001263
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
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