{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000792","ANALYSIS_ID":"AN001261","VERSION":"1","CREATED_ON":"July 12, 2017, 2:45 pm"},

"PROJECT":{"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"},

"STUDY":{"STUDY_TITLE":"Large Untargeted Profiling of Myelin to Enhance Recovery of Function after SCI","STUDY_SUMMARY":"Tissue 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_NAME":"Scarisbrick","FIRST_NAME":"Isobel","ADDRESS":"200 First St. SW, Rochester, Minnesota, 55905, USA","EMAIL":"scarisbrick.isobel@mayo.edu","PHONE":"507-284-0124"},

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"Sample ID":"nhilic-1jun16-016-r002_1",
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"Additional sample data":{"type":"NHILIC"}
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{
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"Sample ID":"nhilic-1jun16-017-r001_1",
"Factors":{"Group":"LFD","rep":"1"},
"Additional sample data":{"type":"NHILIC"}
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{
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"Sample ID":"nhilic-1jun16-017-r002_1",
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{
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"Sample ID":"nhilic-1jun16-018-r001_1",
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"Sample ID":"nhilic-1jun16-018-r002_1",
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{
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{
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{
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"Sample ID":"nhilic-1jun16-020-r001_1",
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"Additional sample data":{"type":"NHILIC"}
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{
"Subject ID":"MS6006-20",
"Sample ID":"nhilic-1jun16-020-r002_1",
"Factors":{"Group":"LFD","rep":"2"},
"Additional sample data":{"type":"NHILIC"}
},
{
"Subject ID":"MS6006-4",
"Sample ID":"philic-31may16-004-r001",
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"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-4",
"Sample ID":"philic-31may16-004-r002",
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"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-5",
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{
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},
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"Subject ID":"MS6006-8",
"Sample ID":"philic-31may16-008-r002",
"Factors":{"Group":"HFHS","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-9",
"Sample ID":"philic-31may16-009-r001",
"Factors":{"Group":"Keto","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-9",
"Sample ID":"philic-31may16-009-r002",
"Factors":{"Group":"Keto","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-10",
"Sample ID":"philic-31may16-010-r001",
"Factors":{"Group":"HFHS","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-10",
"Sample ID":"philic-31may16-010-r002",
"Factors":{"Group":"HFHS","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-11",
"Sample ID":"philic-31may16-011-r001",
"Factors":{"Group":"LFD","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-11",
"Sample ID":"philic-31may16-011-r002",
"Factors":{"Group":"LFD","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-12",
"Sample ID":"philic-31may16-012-r001",
"Factors":{"Group":"HFD","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-12",
"Sample ID":"philic-31may16-012-r002",
"Factors":{"Group":"HFD","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-13",
"Sample ID":"philic-31may16-013-r001",
"Factors":{"Group":"HFHS","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-13",
"Sample ID":"philic-31may16-013-r002",
"Factors":{"Group":"HFHS","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-14",
"Sample ID":"philic-31may16-014-r001",
"Factors":{"Group":"HFD","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-14",
"Sample ID":"philic-31may16-014-r002",
"Factors":{"Group":"HFD","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-15",
"Sample ID":"philic-31may16-015-r001",
"Factors":{"Group":"Keto","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-15",
"Sample ID":"philic-31may16-015-r002",
"Factors":{"Group":"Keto","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-16",
"Sample ID":"philic-31may16-016-r001",
"Factors":{"Group":"HFD","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-16",
"Sample ID":"philic-31may16-016-r002",
"Factors":{"Group":"HFD","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-17",
"Sample ID":"philic-31may16-017-r001",
"Factors":{"Group":"LFD","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-17",
"Sample ID":"philic-31may16-017-r002",
"Factors":{"Group":"LFD","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-18",
"Sample ID":"philic-31may16-018-r001",
"Factors":{"Group":"Keto","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-18",
"Sample ID":"philic-31may16-018-r002",
"Factors":{"Group":"Keto","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-19",
"Sample ID":"philic-31may16-019-r001",
"Factors":{"Group":"HFHS","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-19",
"Sample ID":"philic-31may16-019-r002",
"Factors":{"Group":"HFHS","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-20",
"Sample ID":"philic-31may16-020-r001",
"Factors":{"Group":"LFD","rep":"1"},
"Additional sample data":{"type":"PHILIC"}
},
{
"Subject ID":"MS6006-20",
"Sample ID":"philic-31may16-020-r002",
"Factors":{"Group":"LFD","rep":"2"},
"Additional sample data":{"type":"PHILIC"}
}
],
"COLLECTION":{"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."},

"TREATMENT":{"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)."},

"SAMPLEPREP":{"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."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Agilent 1290 Infinity","COLUMN_NAME":"Waters Acquity BEH Amide (150 x 2.1mm, 1.7um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Agilent 6550 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_RESULTS_FILE":"ST000792_AN001261_Results.txt UNITS:intensity"}

}