8^th International Conference on Human Genetics and Genetic Diseases November 25-26, 2019 Madrid, Spain

Biography:

Ben Borokhovsky is a current MD candidate student at Cooper Medical School of Rowan University. He completed his undergraduate education at Widener University and graduated Summa Cum Laude with a degree in Biochemistry. He did research throughout his entire undergraduate education and has consistently been asked to speak at Widener’s Research Symposium and has even given a keynote presentation on his senior thesis, for which he won a first place award. During his time at Widener University, Ben’s research involved looking at gene regulation of specific SR proteins and the roles they play in lipid metabolism using an alternative splicing model in D. melanogaster to better understand molecular and genetic links to obesity in humans.

Abstract:

The method of gene regulation underlying lipid storage related to obesity is poorly understood, yet alternative splicing (AS) appears to be an important mechanism for proper lipid storage. CPT1 (carnitine palmitoyltransferase 1) is a beta-oxidation enzyme involved in the breakdown of fatty acids. The gene coding for CPT1 is alternatively spliced by the SR protein 9G8 to produce two different products that vary in their activity. My project was to investigate if there was a difference in the AS phenotype among flies with decreased expression of 9G8, the SR protein antagonist RSF1, and the SR protein transporter TRN. First, a quantitative PCR (qPCR) protocol needed to be developed to detect the isoforms of CPT1. cDNA was generated from wild type flies that were fed (0 hrs), starved (65 hrs) or refed to optimize the likelihood that both isoforms would be produced. Following a published protocol, a qPCR procedure was carried out that skipped the elongation step (73) and instead cycled from the standard annealing temperature (60) to the denaturation step (95 ). Additionally, the PCR plasticware resulted in technical difficulties and required troubleshooting. Even though there were difficulties, the qPCR reactions saw good results. The preps were good quality confirmed by qPCR by having rp49 gene come in at 15-17 cycles. The individual isoforms 6A and 6B were both detected at distinct cycle numbers showing that the isoform ratios change depending on which gene KD is occurring and would directly affect lipid storage and contribute to fat reserves. Future directions include running triglyceride (TG) assays to determine if there is a change in TG levels among the 9G8, RSF1, and TRN knockdown flies and then investigate if their splicing patterns changed as well.

Biography:

Litvinova Larisa has completed her MD and currently she is working as Head of Laboratory, Immunology and Cell Biotechnology, Immanuel Kant Baltic Federal University, Kaliningrad, Russian Federation. She has published more than 180 papers in reputed journals.

Abstract:

Introduction: Visfatin has multidirectional effects-pro-inflammatory, stimulating the production of pro-inflammatory cytokines, and neuro-, angioprotective properties. Adipokine has an insulin-mimetic effect and stimulates insulin production. In this regard, the aim of the study was to study the role of visfatin in the development of type 2 diabetes in patients with obesity.

Methodology & Theoretical Orientation: Serum glucose levels were determined using a biochemistry analyzer. Using the method of flow fluorimetry, the content of visfatin, insulin and leptin in the blood plasma was determined using commercial test systems; gene expression level - by PCR.

Findings: In the group of obese patients with type 2 diabetes, the content of leptin, insulin and glucose in the blood plasma was higher, which was the case for patients without type 2 diabetes and control (p <0.05). The level of visfatin in blood plasma was higher than 802.10 (101.0 - 1145.12) pg / ml than in patients with obesity without type 2 diabetes 371.81 (40.83 - 513.58) pg / ml and below the control values (p <0.05). In the group of obese patients with type 2 diabetes, the level of visfatin positively correlated with the level of leptin (r = 0.513) and negatively with the level of insulin (r = -0.435) (p <0.05). However, in patients without type 2 diabetes, the level of visfatin had positive correlations with the content of insulin (r = 0.812) and leptin (r = 0.767) in blood plasma (p <0.05). The expression level of the NAMPT gene (visfatin) was increased in all obese patients with type 2 diabetes in all fat depots, compared with obese patients without it. In obese patients with type 2 diabetes, the level of NAMPT gene expression in subcutaneous adipose tissue was 15 times higher than the control group (p <0.05), while in patients without type 2 diabetes it did not differ from the control, which may indicate the formation of adipokine plasma levels, mainly due to its production of subcutaneous fat depot in patients with type 2 diabetes. The level of NAMPT gene expression in the greater omentum positively correlated with the level of Visfatin in blood plasma in the group of obese patients with type 2 diabetes (r = 0.5 p <0.05). The level of NAMPT gene expression in the mesentery of the small intestine negatively correlated with BMI in the group of obese patients with type 2 diabetes (r = 0.4 p <0.05).

Conclusion & Significance: Thus, visfatin can have different effects on the development of type 2 diabetes in obese patients, mediated by tissue-specific features of its secretion, as indicated by (1) the revealed formation of plasma levels of adipokine due to subcutaneous fat depot in patients with type 2 diabetes; (2) the detected protective effects of visfatin in patients without type 2 diabetes, due to the established positive relationship between the level of visfatin and plasma insulin production in this category of patients.

Biography:

Ms. Ainur Sibagatova is an aspiring researcher. She holds a master degree in health administration. Currently she works in the Gerontology center of Medical Centre Hospital of President’s Affairs Administration of the Republic of Kazakhstan. The interest of her research is the prevention of the age of associated diseases, mainly cardiovascular diseases.

Abstract:

Introduction: Cardiovascular Disease (CVD) is the leading cause of death in Kazakhstan. In the structure of CVD, hypertension takes the first place and leads to remodeling of the myocardium and carotid arteries, which significantly increases cardiovascular risk. A number of studies have been carried out that established the genetic characteristics associated with the risk of hypertension and target organ damage in hypertension, but not in Kazakh population.

The aim of this project is to study the molecular genetic predictors of remodeling of the cardiovascular system in HTN in Kazakh population.

Materials and methods: The sample size is 1000 people. There were 4 groups of 250 hypertensive individuals, age - up to 61 years.

1. Group without remodeling
2. with carotid artery remodeling (CR), intima-media complex (IMC) > 0.9 mm
3. with myocardial remodeling (MR) LVMI ≥115 in men, ≥95 in women, relative wall thickness ≥0.43
4. With remodeling of the myocardium and carotid artery.

Clinical, laboratory, instrumental data were collected. Biological samples were taken for genotyping for 120 SNPs known by association with a thickening of IMC or with left ventricular hypertrophy based on GWAS Catalog - EMBL-EBI and Varsome Clinical and PubMed publications. Genotyping was performed using QuantStudio 12K Flex Real-Time PCR System using array technology.

Preliminary Results: According to a preliminary analysis of 120 SNPs, three polymorphic variants (rs75330306, rs1879734, rs12465515 (p <0.05)) are associated with MR, because of a significant association in groups with MR and in the group RM+CR. rs1879734 and rs12465515 were previously associated with mitral valve prolapse in the European population, and rs75330306 was previously associated with Idiopathic dilated cardiomyopathy in the African American ancestry.

Biography:

Ewa Hordyjewska-Kowalczyk is a young scientist focusing her interest on human genetics and the development of limbs. She has been studying mutations linked to monogenic diseases (Cleidocranial Dysplasia, IFAP syndrome) as well as a complex disorder – Clubfoot.

Abstract:

Introduction: Cleidocranial Dysplasia (CCD) is an autosomal dominant disorder occurring 1 in a million births. The most common phenotypic manifestation of CCD includes hypoplastic or absent clavicles, failure of cranial suture closure and dental anomalies. These defects were linked to mutations in the RUNX2 (Runt-related transcription factor 2), a master regulator of osteoblast differentiation, cartilage, and bone development. Although the gene is known, novel variants are constantly being discovered. The purpose of the study was the functional characterization of six mutations found in Polish patients with CCD.

Methodology & Theoretical Orientation: Members of 11 Polish families were genotyped for variants in the RUNX2 gene. Then to assess the pathogenicity of 6 variants functional studies were carried out, including tests for transactivation potential of RUNX2 mutants and subcellular localization.

Findings: All of the found variants are present in the Runt domain of RUNX2 and are associated with the presence of supernumerary teeth among all patients. What is more, three variants are newly reported missense changes. Furthermore, we showed a significant decrease in the RUNX2 transactivation activity of all mutants, and although only two variants are within Nuclear Localization Signal (NLS), and other two affect the putative NLS, all investigated mutants localize to the cytoplasm.

Conclusion & Significance: The report brings functional evidence on the pathogenicity of found mutations which have a detrimental effect on the RUNX2 and trigger the CCD phenotype. These data not only increase our knowledge about the function of RUNX2 domains but also add to the clinical description and the phenotype/genotype correlation in the patients with these mutations.

Biography:

Abstract:

Introduction: The functioning of mitochondria is disrupted and is associated with a decrease in ATP synthesis and organelle division during obesity under conditions of oxidative stress and increased production of ROS. The aim of the study was to evaluate the expression of genes associated with mitochondrial division / fusion processes in obese patients with type 2 diabetes.

Methodology & Theoretical Orientation: The expression level of genes was determined by real-time PCR. Calculations of the level of relative expression of the studied genes and the significance of differences were determined relative to the control group in the REST program.

Findings: An increase in the level of gene expression of the pro-inflammatory transcription factor NF-kB (2 times relatively healthy donors, p <0.05) in patients with obesity with type 2 diabetes in liver biopsy specimens showed inflammation in the liver parenchyma, whereas in patients without type 2 diabetes of this type, on the contrary, was lower (100 times, p <0.05) of control values. In obese patients with type 2 diabetes, the level of TFAM gene expression increased by 1.3 times (p <0.05), and in obese patients without it, it did not change relative to the control. In patients with obesity with type 2 diabetes, the expression level of the MFN2 and DRP1 gene did not change, and in patients with obesity without it, it decreased by 8.7 times and 100 times, respectively, relative to the control (p <0.05). In this study, in patients with obesity without type 2 diabetes, a decrease in the expression of genes characterizing mitochondrial division and fusion — DRP1 and MFN2 — was recorded; accordingly, a deficiency of proteins involved in mitochondrial fusion may indicate a decrease in cellular respiration and metabolism.

Conclusion & Significance: Thus, an increase in the level of TFAM gene expression in obese patients with type 2 diabetes mellitus indicates an increase in mtDNA transcription, while mitochondrial dysfunction mediated by an increase in the expression level of mitochondrial transcription factor A (TFAM), in the presence of an inflammatory process in the liver in obese patients, may be a factor contributing to the formation of type 2 diabetes.