Marina Vladimirovna Shestakova

Disturbed angiogenic activity of adipose-derived stromal cells obtained from patients with coronary artery disease and diabetes mellitus type 2

BackgroundMultipotent mesenchymal stem/stromal cells (MSC) including adipose-derived stromal cells (ADSC) have been successfully applied for cardiovascular diseases treatment. Their regenerative potential is considered due to the multipotency, paracrine activity and immunologic privilege. However, therapeutic efficacy of autologous MSC for myocardial ischemia therapy is modest. We analyzed if ADSC properties are attenuated in patients with chronic diseases such as coronary artery disease (CAD) and diabetes mellitus type 2 (T2DM).Methods and resultsADSC were isolated from subcutaneous fat tissue of patients without established cardiovascular diseases and metabolic disorders (control group, n = 19), patients with CAD only (n = 32) and patients with CAD and T2DM (n = 28). ADSC phenotype (flow cytometry) was CD90+/CD73+/CD105+/CD45−/CD31− and they were capable of adipogenic and osteogenic differentiation. ADSC morphology and immunophenotype were similar for all patients, but ADSC from patients with CAD and T2DM had higher proliferation activity and shorter telomeres compared to control patients.ADSC conditioned media stimulated capillary-like tubes formation by endothelial cells (EA.hy926), but this effect significantly decreased for patients with CAD (p = 0.03) and with CAD + T2DM (p = 0.017) compared to the control group. Surprisingly we revealed significantly higher secretion of some pro-angiogenic factors (ELISA) by ADSC: vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) for patients with CAD and HGF and placental growth factor (PlGF) for patients with CAD + T2DM. Among angiogenesis inhibitors such as thrombospondin-1, endostatin and plasminogen activator inhibitor-1 (PAI-1) level of PAI-1 in ADSC conditioned media was significantly higher for patients with CAD and CAD + T2DM compared to the control group (p < 0.01). Inhibition of PAI-1 in ADSC conditioned media by neutralizing antibodies partially restored ADSC angiogenic activity (p = 0.017).ConclusionsADSC angiogenic activity is significantly declined in patients with CAD and T2DM, which could restrict the effectiveness of autologous ADSC cell therapy in these cohorts of patients. This impairment might be due to the disturbance in coordinated network of pro- and anti-angiogenic growth factors secreted by ADSC. Changes in ADSC secretome differ between patients with CAD and T2DM and further investigation are necessary to reveal the MSC-involved mechanisms of cardiovascular and metabolic diseases and develop novel approaches to their correction using the methods of regenerative medicine.

Adiponectin and Adiponectin Receptor Gene Variants in Relation to Type 2 Diabetes and Insulin Resistance-Related Phenotypes

BACKGROUND Alterations in adiponectin-mediated pathways are known to be associated with glucose intolerance, insulin resistance (IR), obesity, and type 2 diabetes (T2D) mellitus. Genetic variations in adiponectin (ADIPOQ) and adiponectin 1 and 2 receptor (ADIPOR1 and ADIPOR2) could have effects on IR-related phenotypes and T2D. Here we examine whether the polymorphic markers rs2241766 (ADIPOQ), rs22753738 (ADIPOR1), rs11061971 and rs16928751 (both in ADIPOR2) are implicated in susceptibility to T2D in a Russian population. METHODS The polymorphic markers were genotyped in 129 T2D patients, and 117 non-diabetic controls, by polymerase chain reaction (PCR) restriction fragment length polymorphism approach. In the subjects, biochemical characteristics including serum insulin, plasma glucose and serum lipids/lipoproteins were measured and compared for correlation with the genetic variations studied. RESULTS Allele T of rs11061971 and allele A of rs16928751 showed association with higher risk of diabetes providing odds ratios (OR) of 2.05 (p = 0.0025) and 1.88 (p = 0.018), respectively. Haplotype A-G consisting of allele A of rs11061971 and allele G of rs16928751 was associated with reduced risk of T2D (OR = 0.59, pc = 0.0224). Compared to other variants, diabetic patients double homozygous for A/A of rs16928751 and G/G of rs16928751 had decreased homeostasis model assessment-insulin resistance (pc = 0.0375) and serum triglycerides (pc = 0.0285). CONCLUSIONS The variants of ADIPOR2 confer susceptibility to T2D and are associated with some IR-related phenotypes in the Russian study population.

Contraception in perimenopausal women with diabetes mellitus

Aim. To assess the effect of combined oral contraceptives (COCs) and intrauterine devices (IUDs) on carbohydrate and lipid metabolism and hemostasis in perimenopausal diabetic women. Methods. The open randomized study included a total of 113 diabetic women using COCs with different estrogen/progestogen profiles – ethinylestradiol (EE) 20 μg/desogestrel 150 μg, EE 30 μg/desogestrel 150 μg and EE 30 μg/gestodene 75 μg – and levonorgestrel-releasing or copper IUDs. Average daily insulin requirements, levels of glycosylated hemoglobin, total cholesterol, triglycerides, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol, the state of coagulation hemostatis and fibrinolytic activity were determined at baseline and after 3, 6, 9 and 12 months of contraception. The control group was composed of 40 age-matched diabetic women who did not use any methods of contraception. Results. Neither COCs nor IUDs influenced glycosylated hemoglobin and had little or no influence on the elevation in the requirements for insulin preparations. The majority of the preparations did not exert any unfavorable effect on the blood lipid profile. Taking COCs was accompanied by increased intravascular activation of blood platelets and to a lesser degree by alterations in parameters of hemostatic homeostasis. The use of IUDs had a neutral effect on blood coagulation and fibrinolysis systems. Conclusion. Comparing lipid levels and hemostatic variables as a function of glycosylated hemoglobin level, we conclude that diabetes control has greater influence on these parameters than the type and dose of steroids involved in the contraceptive devices.

National register of diabetes mellitus in Russian Federation.

Clinical and epidemiological monitoring of diabetes in Russian Federation (RF) is carried out by the National diabetes register, which methodological and organizational reference center is Federal Endocrinology Research Centre. Since 2014 initiated the translation the National diabetes register on online software system in order to increase the effectiveness of the register as a scientific and analytical platform. The article include the analysis of DM prevalence, incidence,DM-related mortality,the prevalence of diabetic complications and analysis of effectiveness of diabetes care (included HbA 1c ) and according to the data of online register.

Once-weekly administration of dulaglutide, a glucagon-like peptide-1 receptor agonist, as monotherapy and combination therapy: review of the AWARD studies

For the purpose of exploring the development, pharmacology and clinical trial program related to dulaglutide, we conducted a nonsystematic review of dulaglutide, focusing on the AWARD (Assessment of Weekly Administration of LY2189265 [dulaglutide] in Diabetes Assessment) program of randomized, phase 3 studies. Dulaglutide is a glucagon-like peptide 1 (GLP-1) receptor agonist that causes a variety of antidiabetogenic actions by acting on the incretin system. Steady-state plasma concentrations of dulaglutide are achieved between 2 and 4 weeks following a once-weekly administration. The AWARD 1–6 studies were conducted in patients with different treatment needs, ranging from patients with mild diabetes who can be treated with diet management and exercise to patients for whom target glycemic control cannot be achieved with conventional insulin treatment. Changes in HbA1c from baseline to the primary endpoint assessment (primary efficacy outcome) with dulaglutide were generally dose-dependent and significantly greater than those of active comparators and placebos (AWARD 1–5) or non-inferior to the comparators (AWARD 6). The results of secondary outcome measures demonstrated that glycemic control attained with dulaglutide was sustained for long-term and that a greater proportion of patients treated with dulaglutide achieved a target HbA1c level of <7.0% compared with placebo and/or active comparators. Reduction in fasting serum glucose, glucagon levels and body weight after dulaglutide treatment were noted in most AWARD studies. Although treatment-emergent adverse events were common with dulaglutide, the incidence was similar to that of active comparators in most AWARD studies, and serious adverse events were generally infrequent, except in patients with more severe disease or with prolonged therapy. Hypoglycemic and immunogenic events were also infrequent. In patients with type 2 diabetes, dulaglutide improves glycemic control and leads to clinically useful reduction in body weight in a range of treatment settings.

The PPARγ Pro12Ala variant is associated with insulin sensitivity in Russian normoglycaemic and type 2 diabetic subjects

The second isoform of the PPARγ2 is specific for adipose tissue. In adipocytes, this isoform is involved in the regulation of adipogenesis and lipid storage, insulin and glucose metabolism. Pro12Ala, a missense mutation in exon 2 of PPARG, reduces transcriptional activity of PPARγ2 and is shown to be associated with increased insulin sensitivity and protection from T2D. Previously, this polymorphism has never been assessed in a Russian population for its relationship to T2D, insulin resistance, and diabetes-related metabolic traits. In this study, we tested 588 Russian T2D patients and 597 normoglycaemic controls. Carriers of the Pro12 allele and subjects homozygous for Pro/Pro had significantly increased risk of developing T2D (OR 1.43 and 2.04, respectively). In Pro/Pro homozygotes, adjustment for potential confounding risk factors resulted in reducing the OR value from 2.04 to 1.69, but the association remained significant (p=0.046).The Pro/Pro genotype also showed association with increased levels of fasting insulin (p=0.019) in non-diabetic controls and elevated serum triglycerides (p=0.019) in T2D patients. Compared with other genotypes, non-diabetic and diabetic subjects homozygous for Pro/Pro had a significantly higher HOMA-IR score and reduced ISI value. This observation strongly supports the implication of the PPARG Pro12Ala in insulin resistance and T2D in a Russian population.

Regulation of Adipose Tissue Stem Cells Angiogenic Potential by Tumor Necrosis Factor-Alpha

Tissue regeneration requires coordinated “teamwork” of growth factors, proteases, progenitor and immune cells producing inflammatory cytokines. Mesenchymal stem cells (MSC) might play a pivotal role by substituting cells or by secretion of growth factors or cytokines, and attraction of progenitor and inflammatory cells, which participate in initial stages of tissue repair. Due to obvious impact of inflammation on regeneration it seems promising to explore whether inflammatory factors could influence proangiogenic abilities of MSC. In this study we investigated effects of TNF‐α on activity of adipose‐derived stem cells (ADSC). We found that treatment with TNF‐α enhances ADSC proliferation, F‐actin microfilament assembly, increases cell motility and migration through extracellular matrix. Exposure of ADSC to TNF‐α led to increased mRNA expression of proangiogenic factors (FGF‐2, VEGF, IL‐8, and MCP‐1), inflammatory cytokines (IL‐1β, IL‐6), proteases (MMPs, uPA) and adhesion molecule ICAM‐1. At the protein level, VEGF, IL‐8, MCP‐1, and ICAM‐1 production was also up‐regulated. Pre‐incubation of ADSC with TNF‐α‐enhanced adhesion of monocytes to ADSC but suppressed adherence of ADSC to endothelial cells (HUVEC). Stimulation with TNF‐α triggers ROS generation and activates a number of key intracellular signaling mediators known to positively regulate angiogenesis (Akt, small GTPase Rac1, ERK1/2, and p38 MAP‐kinases). Pre‐treatment with TNF‐α‐enhanced ADSC ability to promote growth of microvessels in a fibrin gel assay and accelerate blood flow recovery, which was accompanied by increased arteriole density and reduction of necrosis in mouse hind limb ischemia model. These findings indicate that TNF‐α plays a role in activation of ADSC angiogenic and regenerative potential. J. Cell. Biochem. 117: 180–196, 2016.

Contrast-induced nephropathy in patients with type 2 diabetes during coronary angiography: Risk-factors and prognostic value

AIMS To determine risk factors, prognostic, value prevention of development of contrast-induced nephropathy (CIN) after percutaneous coronary intervention (PCI) in patients with type 2 diabetes mellitus (T2DM). MATERIALS AND METHODS We have retrospectively analyzed the incidence of CIN developed after PCI in 151 patients T2DM and 50 patients without diabetes. All patients were subjected to thorough clinical examination (including serum creatinine level before and 48 hours after intervention). RESULTS CIN developed more frequently after PCI in patients with T2DM than in patients of the same age without diabetes at the same baseline renal function, volume of contrast media and hydration status. The risk of developing CIN in patients with T2DM is associated with: heart failure, anemia, volume of contrast media, diuretics use in the peri-procedure period, multiple coronary artery disease, need of interventional procedures. TIDM patients with CIN had faster decline of renal function, more often developed cardiovascular diseases and had lower 24 month survival rate. CONCLUSIONS High risk of CIN development and its prognostic significance in patients with T2DM determine the necessity of individually evaluated risks for preventive measures during contrast media interventions.

Prevalence of type 2 diabetes mellitus (T2DM) in the adult Russian population (NATION study).

AIM To estimate type 2 diabetes mellitus (T2DM) prevalence in Russian adults. METHODS NATION is a national, epidemiological, cross-sectional study, conducted in Russia. In adults (aged 20-79 years), recruitment was stratified by age, sex, geographic region and settlement type to obtain a representative sample. Recruitment was in public areas with high numbers of people. T2DM was diagnosed by glycated haemoglobin A1c (HbA1c) levels (diabetes: HbA1c ≥6.5% [≥48mmol/mol]; pre-diabetes: HbA1c ≥5.7 to <6.5% [≥39 to <48mmol/mol]). Socio-demographic and anthropometric data were collected. RESULTS Blood samples from 26,620 subjects were available. Overall, 5.4% were diagnosed with T2DM (previously diagnosed: 2.5%; previously undiagnosed: 2.9%); 19.3% were pre-diabetic. T2DM prevalence increased with age (up to 70 years) and was higher among females than males (6.1% vs. 4.7%, p<0.001). The estimated proportion of subjects with pre-diabetes and T2DM tended to increase with increasing body mass index. T2DM prevalence was higher in rural versus urban populations (6.7% vs. 5.0%, p<0.001). CONCLUSION In the Russian adult population, 19.3% had pre-diabetes, T2DM prevalence was 5.4%, and 54% of subjects with diabetes were previously undiagnosed. These results may help to develop a new T2DM predictive, preventative and management programme in Russia.

The prevalence of type 2 diabetes mellitus in the adult population of Russia (NATION study)

Aim . To estimate the prevalence of type 2 diabetes mellitus (T2DM) in Russian adults. Materials and methods . NATION was a national, epidemiological, observational, cross-sectional study. The sample of adults (20–79 years old) was stratified by age, sex, geographic region and settlement type to obtain a representative sample. Recruitment was performed in public areas with large number of people. T2DM was diagnosed by glycated haemoglobin A1c (HbA1c) levels (diabetes: HbA1c≥6.5%; pre-diabetes: HbA1c≥5.7% to <6.5%). Socio-demographic and anthropometric data were collected. Results . Blood samples from 26,620 subjects were available. Overall, 5.4% were diagnosed with T2DM (2.5% were previously diagnosed and 2.9% were previously undiagnosed); 19.3% were pre-diabetic. T2DM prevalence increased with age (up to 70 years), and no significant difference was revealed between females and males (5.6% vs. 5.1%). The estimated prevalence of pre-diabetes and T2DM tended to increase with increasing BMI. T2DM prevalence varied by geographic region and was higher in rural areas than in urban areas (6.7% vs. 5.0%, p < 0.001). Conclusion . Approximately one in five adult Russians had pre-diabetes, 5.4% had T2DM and about half of the diabetic subjects were previously undiagnosed. These results demonstrate the need for new programs in the Russian Federation to predict, prevent and manage T2DM.