Claire L. McTernan

Resistin, central obesity, and type 2 diabetes

Resistin, an adipocyte-derived cytokine, causes insulin resistance and glucose intolerance in mice. We investigated whether resistin expression was higher in human abdominal adipose tissue than other adipose tissue depots. We extracted RNA from 32 adipose tissue samples (13 subcutaneous abdominal, seven omentum, six thigh, and six breast). Quantitative PCR was used to determine resistin mRNA expression. Resistin mRNA concentrations were similar in both the subcutaneous abdominal and omental depots. The abdominal depots showed a 418% increase in resistin mRNA expression compared with the thigh. Increased resistin expression in abdominal fat could explain the increased risk of type 2 diabetes associated with central obesity.

CA-Repeat Polymorphism in Intron 1 of HSD11B2: Effects on Gene Expression and Salt Sensitivity

Mutations in the HSD11B2 gene encoding the kidney (11-HSD2) isozyme of 11beta-hydroxysteroid dehydrogenase cause apparent mineralocorticoid excess, a form of familial hypertension. Because the hypertension associated with AME is of the salt-sensitive type, it seemed possible that decreases in 11-HSD2 activity might be associated with salt sensitivity. To examine this, Italians with mild hypertension underwent a protocol consisting of a rapid intravenous saline infusion and subsequent furosemide diuresis. To determine whether there were genetic associations between HSD11B2 and salt sensitivity, 198 Italians were genotyped for a CA repeat polymorphism (11 alleles) in the first intron. Increased differences in mean arterial pressure between the sodium loaded and depleted states were correlated with shorter CA repeat length (R=0.214, P=0. 0025). The effect behaved as a recessive trait. This suggested that decreased HSD11B2 expression was associated with shorter CA repeat length. Furthermore, activity of renal 11-HSD2 as measured by an increase in the ratio of urinary-free cortisol/urinary-free cortisone was lower in 33 salt-sensitive subjects (urinary-free cortisol/urinary-free cortisone 0.89+/-0.04 [mean+/-SE]) compared with 34 salt-resistant subjects (0.71+/-0.04, P<0.001). However, when minigenes containing either 14 or 23 CA repeats were transfected into rabbit or human kidney cortical collecting duct cells, the construct with 14 repeats was instead expressed at levels 50% higher than those of the construct with 23 repeats, as determined by reverse transcription-polymerase chain reaction. We conclude that polymorphisms in HSD11B2 and decreased 11-HSD2 activity are associated with sensitivity to sodium loading, but a functional explanation for these associations remains to be elucidated.

Late-Onset Apparent Mineralocorticoid Excess Caused by Novel Compound Heterozygous Mutations in the HSD11B2 Gene

Abstract—Mutations in the gene encoding 11&bgr;-hydroxysteroid dehydrogenase type 2, 11&bgr;-HSD2 (HSD11B2), explain the molecular basis for the syndrome of apparent mineralocorticoid excess (AME), characterized by severe hypertension and hypokalemic alkalosis. Cortisol is the offending mineralocorticoid in AME, as the result of a lack of 11&bgr;-HSD2–mediated cortisol to cortisone inactivation. In this study, we describe mutations in the HSD11B2 gene in 3 additional AME kindreds in which probands presented in adult life, with milder phenotypes including the original seminal case reported by Stewart and Edwards. Genetic analysis of the HSD11B2 gene revealed that all probands were compound heterozygotes, for a total of 7 novel coding and noncoding mutations. Of the 7 mutations detected, 6 were investigated for their effects on gene expression and enzyme activity by the use of mutant cDNA and minigene constructs transfected into HEK 293 cells. Four missense mutations resulted in enzymes with varying degrees of activity, all <10% of wild type. A further 2 mutations generated incorrectly spliced mRNA and predicted severely truncated, inactive enzyme. The mothers of 2 probands heterozygous for missense mutations have presented with a phenotype indistinguishable from “essential” hypertension. These genetic and biochemical data emphasize the heterogeneous nature of AME and the effects that heterozygosity at the HSD11B2 locus can have on blood pressure in later life.

Rosiglitazone inhibits the insulin‐mediated increase in PAI‐1 secretion in human abdominal subcutaneous adipocytes

Objective:  The aim of this study was to investigate the effect of insulin and an insulin‐sensitizing agent, rosiglitazone (RSG), on the production of plasminogen‐activator inhibitor‐1 (PAI‐1) in isolated subcutaneous abdominal adipocytes. Human tissue‐type plasminogen activator (t‐PA) was also measured to assess changes in overall thrombotic risk.

Insulin increases angiotensinogen expression in human abdominal subcutaneous adipocytes

Summary  The renin–angiotensin system is an important regulator of blood pressure, and blockade of this system improves blood pressure in obesity and type 2 diabetes. Recently, components of the system have been described in adipose tissue. However, to date no study has investigated the influence of varying insulin concentrations on angiotensinogen (AGT) protein expression in human subcutaneous abdominal fat. Isolated subcutaneous adipocytes were treated with insulin (1–1000 nm) for 48 h. As part of the studies, a novel AGT antibody was developed and validated by Western blotting and immunohistochemistry. Western blotting was performed on the protein extracted from the adipocytes treated with insulin to determine AGT expression. Increasing doses of insulin raised AGT protein expression in a dose‐dependent manner (control 1.0 ± 0.0 (mean ± s.e.) – protein expression standardized relative to control; 1 nm insulin: 2.64 ± 0.0.32↑***; 100 nm insulin: 4.37 ± 0.57↑***; 1000 nm insulin: 6.50 ± 0.97↑***; ***p < 0.001, n = 3). In conclusion, increasing insulin doses stimulates AGT production. In this study, protein analysis suggests that hyperinsulinaemia may be an important factor in obesity‐related hypertension.

Possible association but no linkage of the HSD11B2 gene encoding the kidney isozyme of 11β-hydroxysteroid dehydrogenase to hypertension in Black people

OBJECTIVE The HSD11B2 (HSD11K) gene encoding the kidney isozyme of 11β‐hydroxysteroid dehydrogenase is mutated in the syndrome of apparent mineralocorticoid excess, an autosomal recessive form of salt‐sensitive hypertension. This gene is thus a logical candidate locus for risk of essential hypertension.

Assessment of the non-HLA-DR-DQ contribution to IDDM1 in British Caucasian families: analysis of LMP7 polymorphisms.

SUMMARY

Analysis of the role of DPB1-encoded amino acids in the genetic predisposition to type I diabetes mellitus

The role of the DPB1 gene in genetic susceptibility to type I diabetes has yet to be elucidated. Studies of DPB1 alleles are conflicting. Analysis at the amino acid level, rather than consideration of allelic polymorphism, has been informative in determining disease susceptibility encoded by the DRB1 and DQ genes. In this study, therefore, amino acid variation at polymorphic sites of the DPbeta peptide chain encoded by the second exon of the DPB1 gene was analyzed in diabetic and control subjects from white Caucasian, North Indian Asian, and Jamaican populations. Human leukocyte antigen genotypes and haplotypes were analyzed using a logistic-regression approach and the data were conditioned for the effects on disease risk of the DRB1, DQA1, and DQB1 genes. Eight DPbeta amino acid residues were significantly associated with type I diabetes independent of DR and DQ (DPbeta 9, 33, 35, 36, 55, 56, 57, and 69). None of these residues, however, correlated consistently with disease risk in all three racial groups. This contrasts with findings for the DRbeta, DQalpha and DQbeta peptide chains, where the identity of the amino acid at particular sites has been found to correlate with predisposition to type I diabetes.