Development of a gas chromatography/mass spectrometry based metabolomics protocol by means of statistical experimental design

Background: Epigenetic regulation may mediate lipotoxic effects on ␤-cells in type 2 diabetes. Results: Lipotoxicity impairs insulin secretion, and alters metabolism, gene expression, and histone marks in INS-1 832/13 ␤-cells. Conclusion: Perturbed insulin secretion results from epigenetic, genetic, and metabolic adaptations to increased fatty acid metabolism in ␤-cells. Significance: Elucidation of the link between lipotoxicity and insulin secretion is crucial for understanding the pathogenesis of type 2 diabetes.

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“…Experimental designs were created, modeled, and evaluated in MODDE 8.0.1 (Umetrics, Umeå, Sweden) as previously described in detail (34).…”

Section: Methodsmentioning

confidence: 99%

Coordinate Changes in Histone Modifications, mRNA Levels, and Metabolite Profiles in Clonal INS-1 832/13 β-Cells Accompany Functional Adaptations to Lipotoxicity

Malmgren

Sartipy

Danielsson

et al. 2013

Journal of Biological Chemistry

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“…Metabolite profiling was performed by GC/MS as previously described in detail (14,15). Briefly, metabolites were extracted from 20 μL serum after addition of 80 μL methanol and a set of stable isotope-labeled internal standards.…”

Section: Methodsmentioning

confidence: 99%

Metabolite Profiling Reveals Normal Metabolic Control in Carriers of Mutations in the Glucokinase Gene (MODY2)

et al. 2013

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Mutations in the gene encoding glucokinase (GCK) cause a mild hereditary form of diabetes termed maturity-onset diabetes of the young (MODY)2 or GCK-MODY. The disease does not progress over time, and diabetes complications rarely develop. It has therefore been suggested that GCK-MODY represents a metabolically compensated condition, but experimental support for this notion is lacking. Here, we profiled metabolites in serum from patients with MODY1 (HNF4A), MODY2 (GCK), MODY3 (HNF1A), and type 2 diabetes and from healthy individuals to characterize metabolic perturbations caused by specific mutations. Analysis of four GCK-MODY patients revealed a metabolite pattern similar to that of healthy individuals, while other forms of diabetes differed markedly in their metabolite profiles. Furthermore, despite elevated glucose concentrations, carriers of GCK mutations showed lower levels of free fatty acids and triglycerides than healthy control subjects. The metabolite profiling was confirmed by enzymatic assays and replicated in a cohort of 11 GCK-MODY patients. Elevated levels of fatty acids are known to associate with β-cell dysfunction, insulin resistance, and increased incidence of late complications. Our results show that GCK-MODY represents a metabolically normal condition, which may contribute to the lack of late complications and the nonprogressive nature of the disease.

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“…Metabolites were extracted from 40 mL of plasma in methanol/water (80:20, volume for volume), as previously described in detail (7,8), but with the inclusion of Val-Tyr-Val, leucine-enkephalin, and reserpine at 0.81 ng/mL as internal standards. After centrifugation, the supernatant was divided into two aliquots for analysis by gas chromatography/ time-of-flight mass spectrometry (GC/TOF-MS) and ultrahigh-performance liquid chromatography/quadrupole-TOF-MS (UHPLC/QTOF-MS).…”

Section: Sample Preparationmentioning

confidence: 99%

Metabolite Profiling of LADA Challenges the View of a Metabolically Distinct Subtype

et al. 2016

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Latent autoimmune diabetes in adults (LADA) usually refers to GAD65 autoantibodies (GADAb)-positive diabetes with onset after 35 years of age and no insulin treatment within the first 6 months after diagnosis. However, it is not always easy to distinguish LADA from type 1 or type 2 diabetes. In this study, we examined whether metabolite profiling could help to distinguish LADA ( = 50) from type 1 diabetes ( = 50) and type 2 diabetes ( = 50). Of 123 identified metabolites, 99 differed between the diabetes types. However, no unique metabolite profile could be identified for any of the types. Instead, the metabolome varied along a C-peptide-driven continuum from type 1 diabetes via LADA to type 2 diabetes. LADA was more similar to type 2 diabetes than to type 1 diabetes. In a principal component analysis, LADA patients overlapping with type 1 diabetes progressed faster to insulin therapy than those overlapping with type 2 diabetes. In conclusion, we could not find any unique metabolite profile distinguishing LADA from type 1 and type 2 diabetes. Rather, LADA was metabolically an intermediate of type 1 and type 2 diabetes, with those patients closer to the former showing a faster progression to insulin therapy than those closer to the latter.

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Development of a gas chromatography/mass spectrometry based metabolomics protocol by means of statistical experimental design

Cited by 26 publications

References 27 publications

Coordinate Changes in Histone Modifications, mRNA Levels, and Metabolite Profiles in Clonal INS-1 832/13 β-Cells Accompany Functional Adaptations to Lipotoxicity

Coordinate Changes in Histone Modifications, mRNA Levels, and Metabolite Profiles in Clonal INS-1 832/13 β-Cells Accompany Functional Adaptations to Lipotoxicity

Metabolite Profiling Reveals Normal Metabolic Control in Carriers of Mutations in the Glucokinase Gene (MODY2)

Metabolite Profiling of LADA Challenges the View of a Metabolically Distinct Subtype

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