Methods and Guidelines for Measurement of Glucagon in Plasma

Holst, Jens J.; Albrechtsen, Nicolai J. Wewer

doi:10.3390/ijms20215416

Cited by 40 publications

(31 citation statements)

References 59 publications

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“…The radioimmunoassay (4305) also measures proglucagon 1–61 because of the identical C-terminus, but this component plays a very minor role under normal circumstances and has glucagon-like bioactivity, if any [ 24 ]. We are therefore convinced that we have used the ideal assay for measurement of glucagon in this study [ 11 ].…”

Section: Discussionmentioning

confidence: 99%

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Genome-wide association study of circulating levels of glucagon during an oral glucose tolerance test

et al. 2021

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Background In order to explore the pathophysiology underlying type 2 diabetes we examined the impact of gene variants associated with type 2 diabetes on circulating levels of glucagon during an oral glucose tolerance test (OGTT). Furthermore, we performed a genome-wide association study (GWAS) aiming to identify novel genomic loci affecting plasma glucagon levels. Methods Plasma levels of glucagon were examined in samples obtained at three time points during an OGTT; 0, 30 and 120 min, in two separate cohorts with a total of up to 1899 individuals. Cross-sectional analyses were performed separately in the two cohorts and the results were combined in a meta-analysis. Results A known type 2 diabetes variant in EYA2 was significantly associated with higher plasma glucagon level at 30 min during the OGTT (Beta 0.145, SE 0.038, P = 1.2 × 10–4) corresponding to a 7.4% increase in plasma glucagon level per effect allele. In the GWAS, we identified a marker in the MARCH1 locus, which was genome-wide significantly associated with reduced suppression of glucagon during the first 30 min of the OGTT (Beta − 0.210, SE 0.037, P = 1.9 × 10–8), equivalent to 8.2% less suppression per effect allele. Nine additional independent markers, not previously associated with type 2 diabetes, showed suggestive associations with reduced glucagon suppression during the first 30 min of the OGTT (P < 1.0 × 10–5). Conclusions A type 2 diabetes risk variant in the EYA2 locus was associated with higher plasma glucagon levels at 30 min. Ten additional variants were suggestively associated with reduced glucagon suppression without conferring increased type 2 diabetes risk.

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Section: Discussionmentioning

confidence: 99%

“…The analytical detection limit was 1 pmol/l, sensitivity around 1 pmol/l and intra- and inter-assay coefficients of variation were < 6% and < 15%, respectively. The applied methodology was recently evaluated [ 11 ]. Glycated haemoglobin A1c (HbA1c) was measured by high-performance liquid chromatography (HPLC) as described previously [ 5 ].…”

Section: Methodsmentioning

confidence: 99%

Genome-wide association study of circulating levels of glucagon during an oral glucose tolerance test

et al. 2021

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“…This indicates that S. epidermidis treatment impacts hyperinsulinemia, a key driver of insulin resistance, metabolic syndrome, and other disorders including cardiomyopathy 24 . Unfortunately we were unsuccessful, despite many different attempts and strategies, to consistently measure serum GLP-1 levels in mice in this experiment or other positive control experiments 25 . Thus, we cannot conclude that the improved health of the animals correlates with increased levels of GLP-1 in vivo and will likely need to move to a larger animal model to address this question.…”

Section: Discussionmentioning

confidence: 97%

Discovery of a bacterial peptide as a modulator of GLP-1 and metabolic disease

Tomaro-Duchesneau

LeValley

Roeth

et al. 2020

Sci Rep

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Early work in rodents highlighted the gut microbiota's importance in metabolic disease, including Type II Diabetes Mellitus (T2DM) and obesity. Glucagon-like peptide-1 (GLP-1), an incretin secreted by L-cells lining the gastrointestinal epithelium, has important functions: promoting insulin secretion, insulin sensitivity, and β-cell mass, while inhibiting gastric emptying and appetite. We set out to identify microbial strains with GLP-1 stimulatory activity as potential metabolic disease therapeutics. Over 1500 human-derived strains were isolated from healthy individuals and screened for GLP-1 modulation by incubating bacterial cell-free supernatants with NCI H716 L-cells. Approximately 45 strains capable of increasing GLP-1 were discovered. All GLP-1 positive strains were identified as Staphylococcus epidermidis by 16S rRNA sequencing. Mass spectrometry analysis identified a 3 kDa peptide, Hld (delta-toxin), present in GLP-1 positive supernatants but absent in GLP-1 neutral supernatants. Studies in NCI-H716 cells and human jejunal enteroids engineered to make more enteroendocrine cells demonstrated that Hld alone is sufficient to enhance GLP-1 secretion. When administered in high-fatfed mice, Hld-producing S. epidermidis significantly reduced markers associated with obesity and T2DM. Further characterization of Hld suggests GLP-1 stimulatory action of Hld occurs via calcium signaling. The presented results identify a novel host-microbe interaction which may ultimately lead to the development of a microbial peptide-based therapeutic for metabolic disease. Metabolic disorders, including Type 2 Diabetes Mellitus (T2DM) and obesity, pose a serious public health concern both nationally and globally. According to the Centers for Disease Control and Prevention (CDC), 9.4% of the population of the United States has Diabetes, including 25.2% of those aged 65 years or older 1. Obesity is also a major health concern, with more than one-third of American adults considered obese 2. Current treatment approaches, including intensive lifestyle modifications, diet intervention, and pharmacologics, have proven unsuccessful in controlling the global increase of metabolic disorders. Therefore, a novel approach to combat metabolic disorders is needed. A number of research groups have recently demonstrated the role of the human gut microbiota in metabolism and metabolic disease, leading to the attempt to develop microbial therapeutics. Bäckhed et al. initially spearheaded this research; using germ-free rodents, it was demonstrated that when the microbiota of conventionally raised animals was transplanted into germ-free rodents, the latter developed an increase in adiposity and insulin resistance 3. Studies by a number of different groups also demonstrated the importance of the microbiota in metabolism, described in a recent review 4. Specifically, the gut microbiota has been shown to play an important role in gut hormone modulation, including GLP-1. Administration of prebiotics, non-digestible food ingredients that stimulate the grow...

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“…Plasma acetaminophen analysis was based on amidase hydrolysis, oxidation, and linkage to tetrahydroquinoline, producing a color shift measured by reflectance photometry (670 nm) (Vitros 5.1 FS; Ortho-Clinical Diagnostics). Plasma glucagon was measured with an in-house radioimmunoassay (antibody 4305) directed against the COOH-terminal ( 38 ), which has been validated thoroughly as recently discussed ( 39 ).…”

Section: Methodsmentioning

confidence: 99%

The Role of Glucagon in the Acute Therapeutic Effects of SGLT2 Inhibition

et al. 2020

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Sodium–glucose cotransporter 2 inhibitors (SGLT2i) effectively lower plasma glucose (PG) concentration in patients with type 2 diabetes, but studies have suggested that circulating glucagon concentrations and endogenous glucose production (EGP) are increased by SGLT2i, possibly compromising their glucose-lowering ability. To tease out whether and how glucagon may influence the glucose-lowering effect of SGLT2 inhibition, we subjected 12 patients with type 2 diabetes to a randomized, placebo-controlled, double-blinded, crossover, double-dummy study comprising, on 4 separate days, a liquid mixed-meal test preceded by single-dose administration of either 1 ) placebo, 2 ) the SGLT2i empagliflozin (25 mg), 3 ) the glucagon receptor antagonist LY2409021 (300 mg), or 4 ) the combination empagliflozin + LY2409021. Empagliflozin and LY2409021 individually lowered fasting PG compared with placebo, and the combination further decreased fasting PG. Previous findings of increased glucagon concentrations and EGP during acute administration of SGLT2i were not replicated in this study. Empagliflozin reduced postprandial PG through increased urinary glucose excretion. LY2409021 reduced EGP significantly but gave rise to a paradoxical increase in postprandial PG excursion, which was annulled by empagliflozin during their combination (empagliflozin + LY2409021). In conclusion, our findings do not support that an SGLT2i-induced glucagonotropic effect is of importance for the glucose-lowering property of SGLT2 inhibition.

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Methods and Guidelines for Measurement of Glucagon in Plasma

Cited by 40 publications

References 59 publications

Genome-wide association study of circulating levels of glucagon during an oral glucose tolerance test

Genome-wide association study of circulating levels of glucagon during an oral glucose tolerance test

Discovery of a bacterial peptide as a modulator of GLP-1 and metabolic disease

The Role of Glucagon in the Acute Therapeutic Effects of SGLT2 Inhibition

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