Summary of Study ST000347
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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000347 |
| Study Title | Metabolomic analysis on samples from rats expressing human amylin (cardiac tissue). |
| Study Summary | Human 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 |
| Department | Sarah W. Stedman Nutrition and Metabolism Center |
| Laboratory | Metabolomics lab |
| Last Name | Ilaiwy |
| First Name | Amro |
| Address | 111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA |
| amroilaiwy@gmail.com, monte_willis@med.unc.edu | |
| Phone | 210-596-0171 |
| Submit Date | 2016-02-18 |
| Study Comments | Cardiac tissue |
| Analysis Type Detail | GC-MS |
| Release Date | 2016-03-03 |
| Release Version | 1 |
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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: | SU000367 |
| 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 |
|---|---|---|
| SA015312 | 32 | HIP |
| SA015313 | 33 | HIP |
| SA015314 | 44 | HIP |
| SA015315 | 22 | HIP |
| SA015316 | 18 | HIP |
| SA015317 | 10 | HIP |
| SA015318 | 23 | HIP |
| SA015319 | 15 | HIP |
| SA015320 | SD1 | WT |
| SA015321 | SD2 | WT |
| SA015322 | 3RT32 | WT |
| SA015323 | 2RT32 | WT |
| SA015324 | 2RT33 | WT |
| SA015325 | SD4 | WT |
| SA015326 | 3RT30 | WT |
| SA015327 | 3RT33 | WT |
| Showing results 1 to 16 of 16 |
Collection:
| Collection ID: | CO000361 |
| Collection Summary: | Cardiac tissue was harvested amd then flash frozen in a liquid nitrogen cooled biopress |
| Sample Type: | Muscle |
Treatment:
| Treatment ID: | TR000381 |
| Treatment Summary: | Fraction of cardiac tissue weighed (25–50 mg wet weight), then the finely cut up tissue quickly added to fresh pre-made buffer (50 % acetyl-nitrile, 50 % water, 0.3% formic acid) at a standard concentration of 25 mg/475 mcl buffer then fully homogenized on ice for 10–25 s and placed on dry ice/stored at - 80C |
Sample Preparation:
| Sampleprep ID: | SP000374 |
| 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 | AN000563 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 6890N |
| Column | Agilent DB5-MS (15m x 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: | CH000400 |
| 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 x 0.25mm,0.25um) |
| Chromatography Type: | GC |
MS:
| MS ID: | MS000499 |
| Analysis ID: | AN000563 |
| Instrument Name: | Agilent 5975 |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
