Summary of study ST000350

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 PR000279. The data can be accessed directly via it's Project DOI: 10.21228/M8VC8G This work is supported by NIH grant, U2C- DK119886.

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Study IDST000350
Study TitleMetabolomic analysis on samples from rats expressing human amylin (plasma).
Study SummaryHuman amylin proteotoxicity impairs protein biosynthesis, and alters major cellular signaling pathways in the heart, brain and liver of humanized diabetic rat model in vivo. The 37 amino acid hormone Amylin is co­secreted with insulin from ß cells in the pancreas. In pre­diabetic and obese humans, chronic amylin hypersecretion parallels the course of disease and is involved in the pathophysiology of beta cell destruction in the pancreas. Recent studies in rats with transgenic expression of beta cell amylin (HIP), we have discovered that human amylin is prone to misfolding and has proteotoxic effects in vivo, resulting in the induction of cell death paralleling the pathophysiology of neurodegenerative disease. These misfolded proteotoxic amylin proteins are found to migrate to both the brain and heart to induce both neurologic deficits and cardiac dysfunction. In the present study, we use non­targeted GC­MS metabolomics analysis to investigate the metabolic consequences of amyloidogenic and cytotoxic amylin oligomers and diabetes in HIP heart, brain, liver, and plasma compared to wild type controls at 1 year of age. We identified that HIP hearts had 45 significantly altered metabolites by t­test (p<0.05) compared to wildtype control hearts (0.1­34.3 fold different,N=8/group). Similarly, we identified 30 metabolites significantly different in the HIP brain by t­test (p<0.05) compared to wildtype control brains (0.2­25.2 fold different (N=~10/group). HIP livers had 58 metabolites significantly altered by t­test (p<0.05) compared to wildtype livers (0.01­99.4 fold different,N=~10/group. Pathway enrichment analysis identified a systemic alteration in protein biosynthesis in the heart and brain of HIP rats compared to wild type controls. Alterations in phenylalanine metabolism and aminoacyl­tRNA biosynthesis were specifically affected in heart and plasma. Tyrosine metabolism is affected across organs, including decreased tyrosine (heart), phenylalanine (heart, liver, brain), and increased fumarate (heart, liver, brain). Increased urea and urea cycle were identified in heart and liver. As protein degradation is a major up­regulator of the urea cycle in human rat diabetic models, these findings suggest a broader connection between amylin, diabetes, protein catabolism, and effects on the urea cycle, which may contribute to the increased morbidity and mortality in diabetics at a multi­system level beyond the effects on glucose metabolism.
Institute
Duke University
DepartmentSarah W. Stedman Nutrition and Metabolism Center
LaboratoryMetabolomics lab
Last NameIlaiwy
First NameAmro
Address111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Emailamroilaiwy@gmail.com, monte_willis@med.unc.edu
Phone210-596-0171
Submit Date2016-02-18
Study CommentsBlood plasma
Analysis Type DetailGC-MS
Release Date2016-03-03
Release Version1
Amro Ilaiwy Amro Ilaiwy
https://dx.doi.org/10.21228/M8VC8G
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000279
Project DOI:doi: 10.21228/M8VC8G
Project Title:Non targeted meatbolomic profiling of transgenic- humanized amylin producing rats
Project Type:GC-MS non targeted analysis
Project Summary:Non targeted metabolomic analysis on samples from rats expressing human amylin.
Institute:University of North Carolina at Chapel Hill
Department:McAllister Heart Institute, Department of Internal medicine
Laboratory:Multiple Centers
Last Name:Ilaiwy
First Name:Amro
Address:111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
Email:amroilaiwy@gmail.com
Phone:210-596-0171
Funding Source:NIH, Fondation Leducq

Subject:

Subject ID:SU000370
Subject Type:Animal
Subject Species:Rattus norvegicus
Taxonomy ID:10116
Species Group:Mammal

Factors:

Subject type: Animal; Subject species: Rattus norvegicus (Factor headings shown in green)

mb_sample_id local_sample_id Genotype
SA01536617HIP
SA01536725HIP
SA0153687HIP
SA01536926HIP
SA015370SRT14HIP
SA01537124HIP
SA01537243HIP
SA01537311HIP
SA01537436HIP
SA01537531HIP
SA0153762RT14WT
SA0153772RT13WT
SA015378SD2WT
SA01537941WT
SA0153802RT15WT
SA0153813RT25WT
SA015382SD1WT
SA015383STRL2WT
SA015384SD4WT
SA0153853RT39WT
Showing results 1 to 20 of 20

Collection:

Collection ID:CO000364
Collection Summary:Plasma was collected then flash frozen in a liquid nitrogen cooled biopress
Sample Type:Blood

Treatment:

Treatment ID:TR000384
Treatment Summary:-

Sample Preparation:

Sampleprep ID:SP000377
Sampleprep Summary:The samples were crash deprotonized by methanol precipitation and spiked with D27-deuterated myristic acid (D27-C14:0) as an internal standard for retention-time locking and dried. The trimethylsilyl-D27-C14:0 standard retention time (RT) was set at 16.727 min. Reactive carbonyls were stabilized at 50C with methoxyamine hydrochloride in dry pyridine. Metabolites were made volatile with TMS groups using N-methyl-N-(trimethylsilyl) trifluoroacetamide or MSTFA with catalytic trimethylchlorosilane at 50C.

Combined analysis:

Analysis ID AN000566
Analysis type MS
Chromatography type GC
Chromatography system Agilent 6890N
Column Agilent DB5-MS (15m × 0.25mm, 0.25um)
MS Type EI
MS instrument type Single quadrupole
MS instrument name Agilent 5975
Ion Mode POSITIVE
Units Peak values (Log transformed)

Chromatography:

Chromatography ID:CH000403
Chromatography Summary:GC/MS methods follow previous studies using a 6890 N GC connected to a 5975 Inert single quadrupole MS (Agilent Technologies, Santa Clara, CA) (Bonikos et al. 1975; Fiehn 2008; Kind et al. 2009). The two wall-coated, open-tubular GC columns connected in series are both from J&W/Agilent (part 122–5512), DB5-MS, 15 meters in length, 0.25 mm in diameter, with an 0.25-l m luminal film. Positive ions generated with conventional electron-ionization at 70 eV are scanned broadly from 600 to 50 m/z in the detector throughout the 45 min cycle time.
Instrument Name:Agilent 6890N
Column Name:Agilent DB5-MS (15m × 0.25mm, 0.25um)
Chromatography Type:GC

MS:

MS ID:MS000502
Analysis ID:AN000566
Instrument Name:Agilent 5975
Instrument Type:Single quadrupole
MS Type:EI
Ion Mode:POSITIVE
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