Genotype-based recall to study metabolic effects of genetic variation: a pilot study of <i>PPARG</i> Pro12Ala carriers

<p><b>Aim:</b> To assess practical implications of genotype-based recall (GBR) studies, an increasingly popular approach for in-depth characterization of genotype–phenotype relationships.</p> <p><b>Methods:</b> We genotyped 2500 participants from the Swedish EpiHealth cohort and considered loss-of-function and missense variants in genes with relation to cardiometabolic traits as the basis for our GBR study. Therefore, we focused on carriers and non-carriers of the <i>PPARG</i> Pro12Ala (rs1801282) variant, as it is a relatively common variant with a minor allele frequency (MAF) of 0.14. It has also been shown to affect ligand binding and transcription, and carriage of the minor allele (Ala12) is associated with a reduced risk of type 2 diabetes. We re-invited 39 Pro12Pro, 34 Pro12Ala, and 30 Ala12Ala carriers and performed detailed anthropometric and serological assessments.</p> <p><b>Results:</b> The participation rates in the GBR study were 31%, 44%, and 40%, and accordingly we included 12, 15, and 13 individuals with Pro12Pro, Pro12Ala, and Ala12Ala variants, respectively. There were no differences in anthropometric or metabolic variables among the different genotype groups.</p> <p><b>Conclusions:</b> Our report highlights that from a practical perspective, GBR can be used to study genotype–phenotype relationships. This approach can prove to be a valuable tool for follow-up findings from large-scale genetic discovery studies by undertaking detailed phenotyping procedures that might not be feasible in large studies. However, our study also illustrates the need for a larger pool of genotyped or sequenced individuals to allow for selection of rare variants with larger effects that can be examined in a GBR study of the present size.</p

Role of peroxisome proliferator-activated receptor gamma Pro12Ala polymorphism in human adipose tissue: assessment of adipogenesis and adipocyte glucose and lipid turnover

<p>The protective mechanisms of peroxisome proliferator-activated receptor gamma (PPARγ) Pro12Ala polymorphism in type 2 diabetes (T2D) are unclear. We obtained subcutaneous adipose tissue (AT) before and 3 h after oral glucose (OGTT) in carriers and non-carriers of the Ala allele (12 Pro/Pro, 15 Pro/Ala, and 13 Ala/Ala). Adipogenesis, adipocyte glucose uptake and lipolysis as well as PPARγ target gene expression were investigated and compared between the genotype groups. During fasting and post-OGTT, neither basal nor insulin-stimulated adipocyte glucose uptake differed between genotypes. Compared to fasting, a decreased hormone-sensitive lipase gene expression in Pro/Pro (p < 0.05) was accompanied with a higher antilipolytic effect of insulin post-OGTT (p < 0.01). The adipocyte size was similar across groups. Preadipocyte differentiation rates between Pro/Pro and Ala/Ala were unchanged. In conclusion, no major differences in AT differentiation, glucose uptake, lipolysis or expression of PPARγ target genes were observed between different PPARγ Pro12Ala genotypes. Albeit small, our study may suggest that other pathways in AT or effects exerted in other tissues might contribute to the Pro12Ala-mediated protection against T2D.</p

A Randomized Controlled Trial of Dapagliflozin Plus Once-Weekly Exenatide Versus Placebo in Individuals with Obesity and Without Diabetes: Metabolic Effects and Markers Associated with Bodyweight Loss

<p><b>Article full text</b></p><p><br></p><p>The full text of this article can be found here<b>. </b><a href="https://link.springer.com/article/10.1007/s13300-018-0449-6">https://link.springer.com/article/10.1007/s13300-018-0449-6</a></p><p></p><p><br></p><p><b>Provide enhanced content for this article</b></p><p><br></p><p>If you are an author of this publication and would like to provide additional enhanced content for your article then please contact <a href="http://www.medengine.com/Redeem/”mailto:[email protected]”"><b>[email protected]</b></a>.</p><p><br></p><p>The journal offers a range of additional features designed to increase visibility and readership. All features will be thoroughly peer reviewed to ensure the content is of the highest scientific standard and all features are marked as ‘peer reviewed’ to ensure readers are aware that the content has been reviewed to the same level as the articles they are being presented alongside. Moreover, all sponsorship and disclosure information is included to provide complete transparency and adherence to good publication practices. This ensures that however the content is reached the reader has a full understanding of its origin. No fees are charged for hosting additional open access content.</p><p><br></p><p>Other enhanced features include, but are not limited to:</p><p><br></p><p>• Slide decks</p><p>• Videos and animations</p><p>• Audio abstracts</p><p> </p><p>• Audio slides</p> <p> </p

Plasma level of human secretagogin was increased as a result of human islet transplant failure in mice.

<p>(a) Plasma glucose levels in human islet-transplanted nu/nu mice. Mice (n = 7) were injected with alloxan to ablate the mouse beta cells, at day -3, followed by transplantation with human islets (1000 IEQ) under the kidney capsule at day 0. Plasma glucose level was monitored over 40 days. (b) Human secretagogin level (c) and human C-peptide level were measured in mouse plasma of both normoglycemic and hyperglycemic mice 40 days after human islet transplantation. Data are presented as mean±SD and statistical differences were calculated using Student's unpaired t-test.</p

Clinical and biochemical characteristics of study participants and correlations between characteristics and the secretagogin (SCGN) level.

<p>Clinical and biochemical characteristics of study participants and correlations between characteristics and the secretagogin (SCGN) level.</p

Secretagogin was released from human beta cells after induction of cellular stress.

<p>(a) Secretagogin release, (b) insulin secretion and (c) intracellular secretagogin level of EndoC-βH1 cells treated for 24h with stress inducers, either tunicamycin (10 μg/ml), thapsigargin (1 μM) or cytokine cocktail (IFN-γ (40 ng/ml), IL1-β (20 ng/ml), TNF-α (40 ng/ml)). All inducers were dissolved in DMSO (1:1000) and control cells were incubated in DMSO (1:1000). Data are presented as mean±SD, statistical differences were calculated using one-way ANOVA analysis, n = 4. **** <i>p</i><0.0001.</p

Abundant expression of secretagogin in human islets but not in exocrine tissue.

<p>Total protein extracts of human islets analyzed by two-dimensional difference in gel electrophoresis (DIGE). Protein spots identified as secretagogin by mass spectrometry (nano LC-FTICR MS/MS) are denoted in red. (b) Total protein extracts of exocrine pancreatic tissue analyzed by DIGE. (c) Western blot analysis of total extracts of human islets and exocrine pancreatic tissue and (d) conditioned media from human islets, immunostained with a polyclonal rabbit anti-secretagogin antibody.</p

Silencing of secretagogin in EndoC cells increased ER stress-induced apoptosis.

<p>Caspase 3/7 activity in EndoC-βH1 cells pretreated (48h) with scrambled control or secretagogin siRNA followed by stress induction by either tunicamycin, thapsigargin or cytokine cocktail (IFN-γ, IL1-β, TNF-α) for 24h. All substances were dissolved in DMSO (1:1000) and control cells were incubated in DMSO (1:1000). Scr. siRNA = Cells treated with scrambled siRNA. siSCGN = Secretagogin knock down by siRNA. Data are presented as mean ± SD and statistical differences were calculated using two-way ANOVA analysis, n = 4. ** <i>p</i> = 0.003; **** <i>p</i>< 0.0001.</p

Demographic information of the donors of pancreatic tissue used for immunohistochemical analysis.

<p>Demographic information of the donors of pancreatic tissue used for immunohistochemical analysis.</p

Design and Optimization of Pyrazinecarboxamide-Based Inhibitors of Diacylglycerol Acyltransferase 1 (DGAT1) Leading to a Clinical Candidate Dimethylpyrazinecarboxamide Phenylcyclohexylacetic Acid (AZD7687)

A new series of pyrazinecarboxamide DGAT1 inhibitors was designed to address the need for a candidate drug with good potency, selectivity, and physical and DMPK properties combined with a low predicted dose in man. Rational design and optimization of this series led to the discovery of compound <b>30</b> (AZD7687), which met the project objectives for potency, selectivity, in particular over ACAT1, solubility, and preclinical PK profiles. This compound showed the anticipated excellent pharmacokinetic properties in human volunteers