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Macro- and microvascular complications of type 2 diabetes (T2D), obesity, and dyslipidemia share common metabolic pathways. Here, using a total of 1,300 metabolites from 996 Qatari adults (57% with T2D) and 1,159 metabolites from an independent cohort of 2,618 individuals from the Qatar BioBank (11% with T2D), we identified 373 metabolites associated with T2D, obesity, retinopathy, dyslipidemia and lipoprotein levels, 161 of which were novel. Novel metabolites included phospholipids, sphingolipids, lysolipids, fatty acids, dipeptides, and metabolites of the urea cycle and xanthine, steroid and glutathione metabolism. The identified metabolites enrich pathways of oxidative stress, lipotoxicity, glucotoxicity and proteolysis. Second, we identified 15 patterns we defined as “metabo-clinical signatures.” These are clusters of T2D patients that group together based on metabolite levels and reveal the same clustering in two or more clinical variables (obesity, LDL, HDL, triglycerides, retinopathy). These signatures revealed metabolic pathways associated with different clinical patterns and identified patients with extreme (very high/low) clinical variables associated with extreme metabolite levels in specific pathways. Among our novel findings are the role of N-acetylmethionine in retinopathy in conjunction with dyslipidemia and the possible roles of N-acetylvaline and pyroglutamine in association with high cholesterol levels and kidney function.

Background: Direct podocyte injury in Diabetic nephropathy (DN) leads to proteinuria, glomerular hypertrophy, and accumulation of extra cellular matrix. By using six publicly available gene expression profiles from early DN, DN, advanced DN, and healthy controls. We have analyzed each of the DN stages against healthy controls and also analyzed the datasets using network inference technique to identify genes playing key role in DN pathogenesis. Methodology: We performed modular repertoire analysis for each dataset separately, then, we performed meta-analysis by merging the datasets to obtain robust differentially expressed genes (DEG). Network inference technique was used to explore and detect novel associations between the DEG and to infer the interaction between the set of genes using mutual information (MI) measure. Statistically significant interactions were selected using a threshold p < 10-7 and visualized using force-directed layout in Cytoscape. MCODE tool was used to find densely connected regions in the network. STRING database utilized for functional enrichment of the clusters in the network and for in silico validation of the gene interactions. Results: A total of 193 samples from the gene expression datasets of human kidney tissues were used, where 79 samples from DN, 21 of advanced DN, 10 early DN, 10 DKD, and 73 from controls. Several key genes have been identified in addition to a highly connected small cluster of genes enriched for podocyte differentiation. Two transcription factors, DACH1 and WT1, are important for podocyte differentiation and proper kidney function have been identified. Also, the expression level of TYRO3 which is a podocyte protective factor increased in early DN but was decreased as the disease progressed, per our analysis. Conclusion: This meta-analysis found some of the key genes associated with DN. The level of contribution and relevance of the identified genes in the pathogenesis of DN should be functionally analyzed. Disclosure A. S. Akil: None. S. Subash padmajeya: None. K. Fakhro: None. T. Habib: None. Funding Qatar National Research Fund (NPRP9-229-3-041)

A number of hyperglycemia- and dyslipidemia- triggered pathways, and numerous protein-encoding genes boosting the diabetic retinopathy (DR) progression. Evolving data suggests vast number of microRNAs(miRNAs) which exhibit no protein-coding capacity, are expressed and play key roles in DR pathogenesis. Purpose: of this pilot is to identify the miRNAs involved in pathways altered in lipid metabolism and dyslipidemia and to explore the association of dyslipidemia with the progression of DR. Methodology: 460 patients with diabetes type 2 (T2D) aged 23-77 years including 37 with DR and 490 matched healthy controls age range 18-74, from Qatar biobank cohort. The circulating miRNA profile was assessed in this pilot study. Logistic regression analysis was performed on the phenotypic data to investigate the level of association between A1C, low-density lipoproteins (LDL) and Triglycerides (TG) covariants and the miRNA expression profile in T2D with and without DR. Results: The most broadly characterized miRNAs in lipid metabolism are miR-33a/b; however, we identified another three miRNAs (miR-133b, miR-142-3p and miR-483-5p) shared between LDL and TG phenotypes. In particular, miR-133b appeared to be directly linked to retinal degeneration identified through our miRNA-target disease interaction network analysis. Our results indicated that all three miRNAs are significantly associated with VEGFR2 mediated vascular permeability, insulin signaling, apoptosis, EGFR, TGF-Beta signaling, cellular senescence and degradation pathways leading to the DR initiation than development. Conclusions: miRNAs are not only small regulators of lipid metabolism, but vital influencers in lipid homeostasis and lipoproteins formation and secretion. Dysregulation of these regulatory elements most likely augment the underlying metabolic flaws perceived in lipid disorders and related microvascular complications. Disclosure A. S. Akil: None. S. Subash padmajeya: None. L. A. Jerman: None. T. Habib: None. A. Al-kurbi: None. E. E. Aliyev: None. M. El anbari: None. K. Fakhro: None. Funding Qatar National Research Fund (NPRP9-229-3-041)

MicroRNAs (miRNAs/miRs) are implicated in the pathogenesis of type 2 diabetes (T2D) and its long-term complications such as retinopathy. miRNAs circulating levels have explored as possible biomarkers for the advancement of T2D. We examined the expression of customized set of miRNAs in T2D Qatari nationals with and without diabetic retinopathy (DR) from Qatar Biobank 1000 Qatar omics cohort. MiR-223 appeared to be significantly upregulated in T2D in comparison to the healthy controls. MiR-223 previously found to play a role in insulin resistant human adipose tissues, human and murine obesity and potential regulators of optic nerve regeneration. To further investigate the potential role of miR-223 in the development of DR, we established a zebrafish (ZF) model to functionally validate this potential correlation. Our aim is to investigate the miR-223 with hyperglycemia on the developing retina in the ZF as a vertebrate model. Mimic miR-223 was injected into single cell ZF embryos and incubated in normal and induced hyperglycemia conditions. We have examined the glucose ratios, eye morphology and vasculature structure in the developing model in comparison to the control groups. To understand the cellular mechanisms involved in DR; histological examination, cell apoptosis and proliferation have been performed and to be evaluated. miR-223 over expression resulted in an alteration of the glucose metabolism within the ZF model. Glucose ratio was significantly increased in ZF embryos injected with mimic miR-223. Increased miR-223 in the developing ZF larvae together with hyperglycemia affected the retinal development. The eye size was significantly reduced with degenerated vasculature and altered eye morphology. In conclusion, we have identified a novel correlation between DR development and miR-223. Targeting miR-223 in T2D patients may serve as promising therapeutic strategy to control DR in at risk T2D patients. Disclosure A. S. Akil: None. S. Da’as: None. L. A. Jerman: None. D. Abdelrahman: None. W. Hasan: None. K. Fakhro: None. Funding Qatar National Research Fund (NPRP9-229-3-041)

Background: A 46-year-old Qatari female with a BMI of 25kg/m2 presented with hyperglycemia (blood glucose 13.5 mmol/l), hyperinsulinemia (21µU/mL) with elevated levels of HbA1c (10.3mmol/l). Biochemical investigations showed raised serum C-peptide (2.02ng/ml) and triglycerides (2.07mmol/L) and uric acid (104mg/dL), with normal BP. The objective of this case study is to validate this variant found through WGS in insulin receptor (INSR) gene that play a major cause of insulin resistant syndrome with different spectrums. Methods: Structural Variants discovered using custom multi-algorithm pipeline developed and optimized to run on HPC for short-read WGS data. In brief, 10 software packages applied (Breakdancer, Breakseq2, CNVnator, Delly, ERDS, Genomestrip, Manta, Speedseq, Svaba, WHAM), to existing short-read data. SVs annotated using AnnotSV and provides us functionally, regulatory and clinically relevant information. SVs analyzed based on their properties like pli-score, overlaps with known databases and OMIM annotations. This deletion was validated by ddPCR and the molecular docking performed by the autodock suit in order to understand the effect of deletion in the interaction between this receptor and insulin. Results: The patient was identified with 7.2k heterozygous deletion in exon 8 of INSR gene which play an essential role in the formation of extracellular alpha subunits in insulin receptor. molecular docking proved the decrease of catalytic activity of INSR and, hence signaling. Considering this and the phenotypic data from Qatar biobank, the patient developed features of type A insulin resistant syndrome as consequence of persistence hyperinsulinemia. The patient also exhibits severe Deterioration of glucose tolerant even with medication and unfortunately developed nephropathy. Conclusion: We highlighted the power of NGS in precise diagnosis and developing a potential and promising targeted therapeutics. Disclosure A.S. Akil: None. E.A.M. Yassin: None. S. Subash Padmajeya: None. L.A. Jerman: None. A. Fadda: None. W.H.O. Aamer: None. E.E. Aliyev: None. K. Fakhro: None.

Background: In the light of growing global epidemic of T2D and related microvascular complications, it is important to find early detection marker for the disease. Circulating miRNAs serve as a potential biomarker for monitering the progression to DR in diabetic individuals. Hypothesis and Aims: miRNAs play a substantial role in metabolic homeostasis through regulation of multiple genes, and could serve as a prognostic biomarker in DR. This study aims to assess a functional role of selected miRNAs in the Qatari population recruited for the Qatar Genome Project through Qatar BioBank. Methods: Plasma samples from 470 T2D subjects with and without DR and 500 matched-healthy controls has been included in this study. Total RNA was extracted and the expression profiling of 56 miRNA that previously reported as a player in the pathogenesis of DR was performed using custom open array panel. The miRNA expression data were normalized using ath-mir159a. Statistical analysis was performed to study the association between the phenotypes (HbA1c, C-peptide, age, BMI) and miRNA. Results: The patients and controls phenotypic and miRNA expression data were analyzed to interepret the differential expression of the miRNA in DR subjects. Initial analysis by applying the generalized additive model reveals that six miRNA (hsa-miR-27a, hsa-miR-376c, hsa-miR-92a, hsa-miR-1 hsa-miR-223, and mmu-miR-451) were found to be significantly associated with DR (p>0.05). Furthermore, two differentially expressed miRNAs, (hsa-miR-1274A and hsa-miR-24) found to be associated with higher level of C-peptide in DR patients, which suggest the correlation of these miRNAs with the glucose hemostasis and DR. Conclusion: The etiology of DR is unclear, and present treatments have limited effectiveness. The biomarkers and downstream functional studies are required to deepen the understanding of this chronic diseases and its complications. Disclosure A.S. Akil: None. S. Subash Padmajeya: None. L.A. Jerman: None. A. Al-Kurbi: None. A.M. Hussein: None. A. Hardikar: None. M. Joglekar: None. T. Habib: None. M. El Anbari: None. K. Fakhro: None.

Background: According to WHO, Qatar is one of the countries most affected by type 2 diabetes T2D worldwide. Despite current treatments, patients suffering from diabetes are still at risk of developing microvascular complications. Of which, Diabetic Retinopathy DR is a debilitating microvascular complication, which leads to blindness, if left untreated. Hypothesis and Aims: the genetic variants SNPs, circulating non-coding RNAs, mRNAs, and proteins, detected in diabetic individuals progressing to DR are accurate biomarkers for DR detection, progression and can be utilized effectively in future screening programs for at-risk individuals as well as for monitoring the outcome of interventions/treatments for DR. The aim is to investigate the SNPs using WGS data, and assess microRNA, mRNA and protein signature of DR progression in T2D patients with and without DR comparing to the healthy controls. Methodology: plasma samples, WGS data of 500 T2D with and without DR and 500 healthy controls subjects were included in this study. Retinal scans are available for all the study participants to assess and classify the DR stages. Total RNA was extracted from plasma samples, microRNA customes assays profiling, proteins and mRNAs are currently at the final stages of analysis and will be ready at the time of the presentation. Preliminary Results: our preliminary analyses through genome-and proteome-wide discovery studies using cellular systems and clinical samples, identified an omics signature of DR comprising of: four mRNAs of endothelial stress, 45 microRNAs of progressing to DR, eight proteins of proliferative DR vs. no DR and candidate SNPs identified in a Qatari cohort that are associated with DR progression. Conclusion: This information will significantly impact the development of newer analytical tools and better therapies that will allow early prediction and treatment of DR, thereby reducing the burden of sight threatening diabetes complications in Qatar. Disclosure S. Subash Padmajeya: None. L. Jerman: None. A. Al-Kurbi: None. K. Fakhro: None. A.J. Jenkins: Advisory Panel; Self; Abbott, Australian Diabetes Society, Medtronic. Research Support; Self; Abbott, GlySens Incorporated, Medtronic, Mylan. Speaker's Bureau; Self; Eli Lilly and Company, Novo Nordisk Inc. A. Hardikar: None. A.S. Akil: None. Funding Qatar National Research Fund