ИЗУЧЕНИЕ АССОЦИАЦИИ ПОЛИМОРФИЗМА RS5219 ГЕНА KCNJ11 С РИСКОМ РАЗВИТИЯ САХАРНОГО ДИАБЕТА 2-ГО ТИПА

Background: Type 2 diabetes mellitus (T2DM) represents from 90 to 95% of all diabetes and usually occurs in obese individuals above 40 years of age, is highly prevalent, associated with high morbidity and mortality from complications involving, first of all, the cardiovascular system. The risk of T2DM is determined by combined effects of genetic and environmental factors. Genes associated with T2DM have been identified, including the gene of ATPdependent potassium channel (KCNJ11); the prevalence of its polymorphisms may have some regional characteristics.Aim: To study an association between rs5219 KCNJ11  gene polymorphisms and the risk of T2DM in the population of the Moscow Region.Materials and methods: The study involved 1050  subjects, including 311  men and 739 women, 139 of whom (17 men and 122 women) had T2DM. Genotyping of rs5219 KCNJ11 gene polymorphisms was performed with the use of allele-specific amplification, the real-time detection and TaqMan-probes complementary to the DNA polymorphism sites.Results: The analysis of rs5219 KCNJ11 polymorphism frequencies showed that 14.2% of patients had TT genotype, 44.8 – CT genotype, and 41.1% – normal (wild) CC genotype. The prevalence of the mutant T allele was 36.6%, that of the C allele – 63.4%. The frequency of the mutant T allele in patients with obesity (body mass index≥30  kg/m²) was not significantly different from that in patients without obesity (body mass index30 kg/m²) (38.8% and 35.7%, respectively, odds ratio (OR) 1.14, 95%  confidence interval (CI) 0.907–1.439, p=0.26). At the same time, energy expenditure at rest per kg of lean body mass was significantly lower in men who have rs5219 KCNJ11 gene polymorphism, both in homoand heterozygotes. The frequency of the T allele and TT genotype in diabetic patients was higher than in the control group. An association between TT genotype and the risk of T2DM was found (OR  2.35, CI 1.018–5.43, p=0.04).Conclusion: In the population of the Moscow Region, gene polymorphism rs5219 KCNJ11 contributes to the risk of developing T2DM which is most obvious and statistically significant in homozygotes.Актуальность. Сахарный диабет (СД) 2-го типа составляет 90–95%  всех случаев СД, как правило, развивается у  людей старше 40  лет, страдающих ожирением, отличается высокой распространенностью, заболеваемостью и  смертностью от осложнений, в  первую очередь со стороны сердечно-сосудистой системы. Риск развития СД 2-го  типа определяется совместным действием генетических и  средовых факторов. Идентифицированы гены, ассоциированные с  СД 2-го  типа, к  ним относится ген АТФ-зависимого калиевого канала (KCNJ11), распространенность полиморфизма которого имеет региональные особенности. Цель  – изучение ассоциации полиморфизма rs5219 гена KCNJ11 с риском развития СД 2-го типа у жителей Московского региона.Материал и  методы. Обследованы 1050 человек, из них 311 мужчин и 739 женщин. СД 2-го типа имели 139 человек (17  мужчин и  122  женщины). Генотипирование полиморфизма rs5219 гена KCNJ11 проводили с  применением аллель-специфичной амплификации с  детекцией результатов в  режиме реального времени и использованием TaqMan- зондов, комплементарных полиморфным участкам дезоксирибонуклеиновой кислоты.Результаты. Анализ частоты встречаемости полиморфизма rs5219 гена KCNJ11 показал, что 14,2%  обследованных имели генотип  ТТ, 44,8% – генотип СТ, 41,1% – нормальный (дикий) генотип СС. Распространенность мутантного аллеля Т составляла 36,6%, аллеля С  – 63,4%. Частота встречаемости мутантного аллеля  Т статистически значимо не отличалась у  об- следованных с  ожирением (индекс массы тела≥30  кг/м²) от обследованных с  индексом массы тела30  кг/м² (38,8%  против  35,7%), отношение шансов (ОШ) 1,14, 95% доверительный интервал (ДИ) 0,907–1,439; p=0,26. Вместе с тем величина энерготрат в покое, рассчитанная на 1  кг мышечной массы тела, была достоверно ниже у  мужчин, имеющих полиморфизм rs5219 гена KCNJ11, как при гомо-, так и при гетерозиготном типе. Частота встречаемости аллеля  Т и генотипа ТТ у пациентов с СД 2-го типа была выше, чем в  группе сравнения. Выявлена ассоциация генотипа ТТ с  риском развития СД 2-го  типа (ОШ  2,35, 95%  ДИ  1,018–5,43; p=0,04).Заключение. У  жителей Московского региона полиморфизм rs5219 гена KCNJ11 (аллель ри- ска  Т) вносит свой вклад (наиболее выраженный и статистически значимый при гомозиготном носительстве аллеля Т) в риск развития СД 2-го типа

Markers of combined aerogenic exposure to metal oxides and transformed plasma proteomic profiles in children

Changes in homeostatic balance of the body, primarily at the cellular-molecular level, are a relevant research object in fundamental and applied studies. They can be eligible indicators for predicting negative effects under exposure to chemical risk factors. The aim of this study was to substantiate markers of a transformed plasma proteomic profile in children. These markers should have prognostic value and an evidence-based association with combined aerogenic exposure to metal oxides (copper and nickel oxides used as an example). We propose an innovative methodical approach based on plasma proteomic profiling that includes the following: identification of identical proteins and genes encoding their expression; quantification of indicators within the ‘identical protein – a chemical concentration in blood’ system; prediction of negative effects as per indicators of homeostasis destabilization at the cellular-molecular level under chronic aerogenic exposure to chemicals. The proposed algorithm was tested by comparing changed proteins and peptides identified in plasma proteomic profiles of children exposed simultaneously to nickel and copper oxides in ambient air in actual conditions and small rodents under experimental combined and isolated exposure to the analyzed chemicals in levels equal to real ones. Long-term aerogenic exposure simultaneously to copper and nickel oxides was established to create elevated nickel and copper levels in blood of exposed children substantiated as markers of exposure. They were up to 2.4 times higher against the same indicators in unexposed children and reference levels as well. The results of field observations were verified by elevated levels of the same chemicals in blood under experimental modelling of an equivalent combined exposure performed on biological models. APOBEC1 complement factor (the А1CF gene) was substantiated as an identical proteomic marker based on plasma proteomic profiling in experimental and field investigations. It has an evidence-based association with markers of exposure (nickel and copper simultaneously identified in blood). Lower expression of this protein under persistent combined aerogenic exposure to nickel and copper oxides makes it possible to predict such a negative effect as modification of low density lipoproteins with further induction of atherosclerotic changes in vessels, the latter being a risk factor of cardiovascular diseases

Immunogenic cell death induced by a new photodynamic therapy based on photosens and photodithazine

Background: Anti-cancer therapy is more successful when it can also induce an immunogenic form of cancer cell death (ICD). Therefore, when developing new treatment strategies, it is extremely important to choose methods that induce ICD and thereby activate anti-Tumor immune response leading to the most effective destruction of tumor cells. The aim of this work was to analyze whether the clinically widely used photosensitizers, photosens (PS) and photodithazine (PD), can induce ICD when used in photodynamic therapy (PDT). Methods: Cell death in murine glioma GL261 or fibrosarcoma MCA205 cells was induced by PS-or PD-PDT and cell death was analyzed by MTT or flow cytometry. Intracellular distribution of PS and PD was studied by using the laser scanning microscope. Calreticulin exposure and HMGB1 and ATP release were detected by flow cytometry, ELISA and luminescence assay, respectively. Immunogenicity in vitro was analyzed by co-culturing of dying cancer cells with bone-marrow derived dendritic cells (BMDCs) and rate of phagocytosis and maturation (CD11c+CD86+, CD11c+CD40+) of BMDCs and production of IL-6 in the supernatant were measured. In vivo immunogenicity was analyzed in mouse tumor prophylactic vaccination model. Results: We determined the optimal concentrations of the photosensitizers and found that at a light dose of 20 J/cm2 (\u3bbex 615-635 nm) both PS and PD efficiently induced cell death in glioma GL261 and fibrosarcoma MCA205 cells. We demonstrate that PS localized predominantly in the lysosomes and that the cell death induced by PS-PDT was inhibited by zVAD-fmk (apoptosis inhibitor) and by ferrostatin-1 and DFO (ferroptosis inhibitors), but not by the necroptosis inhibitor necrostatin-1 s. By contrast, PD accumulated in the endoplasmic reticulum and Golgi apparatus, and the cell death induced by PD-PDT was inhibited only by z-VAD-fmk. Dying cancer cells induced by PS-PDT or PD-PDT emit calreticulin, HMGB1 and ATP and they were efficiently engulfed by BMDCs, which then matured, became activated and produced IL-6. Using dying cancer cells induced by PS-PDT or PD-PDT, we demonstrate the efficient vaccination potential of ICD in vivo. Conclusions: Altogether, these results identify PS and PD as novel ICD inducers that could be effectively combined with PDT in cancer therapy