Summary of Study ST000648
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 PR000460. The data can be accessed directly via it's Project DOI: 10.21228/M8M027 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000648 |
| Study Title | Effects of NO Donor Therapy on the Dystrophic Mouse Muscle Amino Acids (part IV) |
| Study Summary | For this aim, we will only use male mdx mice. We will study three groups treated for 7 days with vehicle, naproxcinod (i.e., NO-naproxen), or naproxen (n = 10 each group). Two hours after the final treatment, half the mice in each group will be run to exhaustion on a treadmill. The heart and gastrocnemius, soleus, and quadriceps muscles of one hindlimb will be sent to the Mayo Clinic Metabolomics Resource Core. The heart and quadriceps muscle will be used for untargeted metabolomics profiling (LC/MS) while the gastrocnemius and soleus muscles will be used for targeted analyses of amino acids plus amino metabolites, non-esterified fatty acids, and citric acid cycle intermediates. |
| Institute | Mayo Clinic |
| Last Name | Thomas |
| First Name | Gail |
| Address | Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033 |
| gthomas4@hmc.psu.edu | |
| Phone | 717-531-0003, ext. 287087 |
| Submit Date | 2017-06-23 |
| Analysis Type Detail | LC-MS |
| Release Date | 2019-07-17 |
| Release Version | 1 |
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Project:
| Project ID: | PR000460 |
| Project DOI: | doi: 10.21228/M8M027 |
| Project Title: | The dystrophic muscle metabolome: effects of exercise and NO donor therapy |
| Project Summary: | In Duchenne and Becker muscular dystrophy (DMD, BMD), loss of the cytoskeletal protein dystrophin weakens the sarcolemma and disrupts cellular signaling, rendering the diseased muscles susceptible to contractioninduced damage. We and others have shown that loss of neuronal nitric oxide synthase (nNOSμ) from the sarcolemma of dystrophin-deficient muscle causes functional muscle ischemia during exercise due to unopposed sympathetic vasoconstriction, thereby exacerbating fatigue and injury of the diseased muscles. Genetic and pharmacologic strategies targeting nNOSμ-NO signaling ameliorate functional muscle ischemia, as well as many other features of the dystrophic phenotype in the mdx mouse model of DMD/BMD. These findings suggest that the therapeutic benefit of NO likely extends beyond its vascular effects. A growing body of evidence indicates that NO directly influences muscle metabolism through effects on glucose transport as well as mitochondrial biogenesis and function. Both nNOS-/- mice and mdx mice exhibit muscle mitochondrial dysfunction, decreased resistance to fatigue, and exercise-induced muscle injury, suggesting a causal role of nNOSμ-NO deficiency. However, the specific metabolic changes resulting from reduced NO signaling that might render dystrophic muscle susceptible to fatigue and use-dependent injury remain poorly defined. Therefore, the goal of this pilot metabolomics study is to identify the unique biochemical profiles of skeletal and cardiac muscles of mdx mice to gain further mechanistic insight into the pathophysiological role of NO deficiency in muscular dystrophy. In Aim 1, we will characterize the skeletal and cardiac muscle metabolomes of mdx and nNOS-/- mice at rest and following a single bout of treadmill exercise with the goal of discovering common metabolic signatures caused by loss of NO signaling. In Aim 2, we will evaluate the potential of a NO donor drug that is under development as a therapeutic for DMD/BMD to improve the skeletal and cardiac muscle metabolomes in mdx mice. As a result of this pilot study, we hope to gain new understanding of the metabolic derangements in dystrophin-deficient muscle, insight into the therapeutic effects of NO replacement, and to identify new pathogenic mechanisms and putative therapeutic targets that will form the basis of future grant applications. |
| Institute: | Mayo Clinic |
| Last Name: | Thomas |
| First Name: | Gail |
| Address: | Penn State Hershey Heart and Vascular Institute Penn State College of Medicine 500 University Drive, MC H047 Hershey, PA 17033 |
| Email: | gthomas4@hmc.psu.edu |
| Phone: | 717-531-0003, ext. 287087 |
Subject:
| Subject ID: | SU000671 |
| 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 | Time point | Grouping |
|---|---|---|---|
| SA036529 | ms5862-6 | Post Run | RUN-Naproxcinod |
| SA036530 | ms5862-8 | Post Run | RUN-Naproxcinod |
| SA036531 | ms5862-10 | Post Run | RUN-Naproxcinod |
| SA036532 | ms5862-7 | Post Run | RUN-Naproxcinod |
| SA036533 | ms5862-9 | Post Run | RUN-Naproxcinod |
| SA036534 | ms5862-18 | Post Run | RUN-Naproxen |
| SA036535 | ms5862-20 | Post Run | RUN-Naproxen |
| SA036536 | ms5862-19 | Post Run | RUN-Naproxen |
| SA036537 | ms5862-17 | Post Run | RUN-Naproxen |
| SA036538 | ms5862-16 | Post Run | RUN-Naproxen |
| SA036539 | ms5862-30 | Post Run | RUN-Vehicle |
| SA036540 | ms5862-27 | Post Run | RUN-Vehicle |
| SA036541 | ms5862-29 | Post Run | RUN-Vehicle |
| SA036542 | ms5862-28 | Post Run | RUN-Vehicle |
| SA036543 | ms5862-26 | Post Run | RUN-Vehicle |
| SA036544 | ms5862-3 | Sedentary | SED-Naproxcinod |
| SA036545 | ms5862-2 | Sedentary | SED-Naproxcinod |
| SA036546 | ms5862-4 | Sedentary | SED-Naproxcinod |
| SA036547 | ms5862-5 | Sedentary | SED-Naproxcinod |
| SA036548 | ms5862-1 | Sedentary | SED-Naproxcinod |
| SA036549 | ms5862-13 | Sedentary | SED-Naproxen |
| SA036550 | ms5862-14 | Sedentary | SED-Naproxen |
| SA036551 | ms5862-15 | Sedentary | SED-Naproxen |
| SA036552 | ms5862-12 | Sedentary | SED-Naproxen |
| SA036553 | ms5862-11 | Sedentary | SED-Naproxen |
| SA036554 | ms5862-21 | Sedentary | SED-Vehicle |
| SA036555 | ms5862-22 | Sedentary | SED-Vehicle |
| SA036556 | ms5862-23 | Sedentary | SED-Vehicle |
| SA036557 | ms5862-24 | Sedentary | SED-Vehicle |
| SA036558 | ms5862-25 | Sedentary | SED-Vehicle |
| Showing results 1 to 30 of 30 |
Collection:
| Collection ID: | CO000665 |
| Collection Summary: | Blood and Tissue Harvesting: Mice will be euthanized and blood will be taken by cardiac puncture, centrifuged, and the plasma will be stored at -80ºC. The heart, diaphragm, and muscles of both hindlimbs (gastrocnemius, soleus, quadriceps) will be dissected and snap frozen in liquid nitrogen. Tibialis anterior and plantaris muscles from both hindlimbs also will be dissected and one set will be snap frozen in liquid nitrogen while the other set will be mounted in OCT and frozen in isopentane cooled by liquid nitrogen. Cryosections will be used to evaluate disease activity and exercise-induced muscle injury by staining with: (a) hematoxylin and eosin to assess gross morphology, cellular infiltration, and necrosis, (b) anti-F4/80 to label macrophages, and (c) anti-IgG or IgM to label damaged muscle fibers. |
| Sample Type: | Heart tissue |
Treatment:
| Treatment ID: | TR000685 |
| Treatment Summary: | Studies will be performed in 12-16 week old male mdx mice, C57BL10 control mice, and nNOS-/- mice obtained from Jackson Laboratory. All protocols will be approved by the Penn State College of Medicine Institutional Animal Care and Use Committee. Drug Treatment: Mice will be treated with vehicle, naproxcinod (20 mg/kg/day) or equimolar naproxen (12.5mg/kg/day) once a day for 7 consecutive days. To avoid the stress of oral gavage or ip injections in mdx mice, drug or vehicle will be administered in a small volume of peanut butter. Most mice readily consume the dosed peanut butter within 30 min. Terminal experiments will be performed 2 hours after the final dose is consumed. |
Sample Preparation:
| Sampleprep ID: | SP000678 |
| Sampleprep Summary: | mouse gastrocnemius amino acids |
Combined analysis:
| Analysis ID | AN000980 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity |
| Column | Waters Acquity BEH C18 (150 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Thermo Quantum Ultra |
| Ion Mode | POSITIVE |
| Units | picomoles/mg |
Chromatography:
| Chromatography ID: | CH000705 |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity BEH C18 (150 x 2.1mm,1.7um) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000875 |
| Analysis ID: | AN000980 |
| Instrument Name: | Thermo Quantum Ultra |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
