Multi-omics approaches, which integrate genomics, transcriptomics, proteomics, and metabolomics, have emerged as powerful tools in the diagnosis of rare diseases. We used untargeted metabolomics and whole-genome sequencing (WGS) to gain a more comprehensive understanding of a rare disease with a complex presentation affecting female twins from a consanguineous family. The sisters presented with polymicrogyria, a Dandy–Walker malformation, respiratory distress, and multiorgan dysfunctions. Through WGS, we identified two rare homozygous variants in both subjects, a pathogenic variant in ADGRG1(p.Arg565Trp) and a novel variant in CNTNAP1(p.Glu910Val). These genes have been previously associated with autosomal recessive polymicrogyria and hypomyelinating neuropathy with/without contractures, respectively. The twins exhibited symptoms that overlapped with both of these conditions. The results of the untargeted metabolomics analysis revealed significant metabolic perturbations relating to neurodevelopmental abnormalities, kidney dysfunction, and microbiome. The significant metabolites belong to essential pathways such as lipids and amino acid metabolism. The identification of variants in two genes, combined with the support of metabolic perturbation, demonstrates the rarity and complexity of this phenotype and provides valuable insights into its underlying mechanisms.

Mandibulofacial dysostosis with microcephaly (MFDM) is a rare genetic disorder inherited in an autosomal dominant pattern. Major characteristics include developmental delay, craniofacial malformations such as malar and mandibular hypoplasia, and ear anomalies. Here, we report a 4.5-yr-old female patient with symptoms fitting MFDM. Using whole-genome sequencing, we identified a de novo start-codon loss (c.3G > T) in the EFTUD2 We examined EFTUD2 expression in the patient by RNA sequencing and observed a notable functional consequence of the variant on gene expression in the patient. We identified a novel variant for the development of MFDM in humans. To the best of our knowledge, this is the first report of a start-codon loss in EFTUD2 associated with MFDM.

While de novo mutations (DNMs) are key to genetic diversity, they are also responsible for a high number of rare disorders. To date, no study has systematically examined the rate and distribution of DNMs in multiplex families in highly consanguineous populations. Leveraging WGS profiles of 645 individuals in 146 families, we implemented a combinatorial approach using 3 complementary tools for DNM discovery in 353 unique trio combinations. We found a total of 27,168 DNMs (median: 70 single-nucleotide and 6 insertion-deletions per individual). Phasing revealed around 80% of DNMs were paternal in origin. Notably, using whole-genome methylation data of spermatogonial stem cells, these DNMs were significantly more likely to occur at highly methylated CpGs (OR: 2.03; p value = 6.62 × 10−11). We then examined the effects of consanguinity and ethnicity on DNMs, and found that consanguinity does not seem to correlate with DNM rate, and special attention has to be considered while measuring such a correlation. Additionally, we found that Middle-Eastern families with Arab ancestry had fewer DNMs than African families, although not significant (p value = 0.16). Finally, for families with diseased probands, we examined the difference in DNM counts and putative impact across affected and unaffected siblings, but did not find significant differences between disease groups, likely owing to the enrichment for recessive disorders in this part of the world, or the small sample size per clinical condition. This study serves as a reference for DNM discovery in multiplex families from the globally under-represented populations of the Middle-East.

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)

Studies assessing the impact of amylase genes copy number (CN) on adiposity report conflicting findings in different global populations, likely reflecting the impact of ancestral and ethnic-specific environment and lifestyle on selection at the amylase loci. Here, we leverage population size and detailed adiposity measures from a large population biobank to resolve confounding effects and determine the relationship between salivary (AMY1) and pancreatic (AMY2A) amylase genes CN and adiposity in 2935 Qatari individuals who underwent whole-genome sequencing (WGS) as part of the Qatar Genome Programme. We observe a negative association between AMY1 CNs and trunk fat percentage in the Qatari population (P = 7.50 × 10−3) and show that Qataris of Arab descent have significantly lower CN at AMY1 (P = 1.32 × 10−10) as well as less favorable adiposity and metabolic profiles (P < 1.34 × 10−8) than Qataris with Persian ancestry. Indeed, lower AMY1 CN was associated with increased total and trunk fat percentages in Arabs (P < 4.60 × 10−3) but not in Persians. Notably, overweight and obese Persians reported a significant trend towards dietary restraint following weight gain compared to Arabs (P = 4.29 × 10−5), with AMY1 CN showing negative association with dietary self-restraint (P = 3.22 × 10−3). This study reports an association between amylase gene CN and adiposity traits in a large Middle Eastern population. Importantly, we leverage rich biobank data to demonstrate that the strength of this association varies with ethnicity, and may be influenced by population-specific behaviors that also contribute to adiposity traits.

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)

Type 1 diabetes (T1D) is an autoimmune condition where the body’s immune cells destroy their insulin-producing pancreatic beta cells leading to dysregulated glycaemia. Individuals with T1D control their blood glucose through exogenous insulin replacement therapy, often using multiple daily injections or pumps. However, failure to accurately mimic intrinsic glucose regulation results in glucose fluctuations and long-term complications impacting key organs such as the heart, kidneys, and/or the eyes. It is well established that genetic and environmental factors contribute to the initiation and progression of T1D, but recent studies show that epigenetic modifications are also important. Here, we discuss key epigenetic modifications associated with T1D pathogenesis and discuss how recent research is finding ways to harness epigenetic mechanisms to prevent, reverse, or manage T1D.

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: 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: 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

This paper depicts the first report from an Indian population on the association between the variant Arg399Gln of XRCC1 locus in the DNA repair system and schizophrenia, the debilitating disease that affects 1% of the world population. Genotypic analysis of a total of 523 subjects (260 patients and 263 controls) revealed an overwhelming presence of Gln399Gln in the case subjects against the controls (P < 0.0068), indicating significant level of association of this nsSNP with schizophrenia; the Gln399 allele frequency was also perceptibly more in cases than in controls (p < 0.003; OR = 1.448). The results of the genotypic studies were further validated using pathogenicity and stability prediction analysis employing computational tools [I-Mutant Suite, iStable, PolyPhen2, SNAP, and PROVEAN], with a view toassess the magnitude of deleteriousness of the mutation. The pathogenicity analysis reveals that the nsSNP could be deleterious inasmuch as it could affect the functionality of the gene, and interfere with protein function. Molecular dynamics simulation of 60ns was performed using GROMACS to analyse structural change due to a mutation (Arg399Gln) that was never examined before. RMSD, RMSF, hydrogen bonds, radius of gyration and SASA analysis showedthe existence of asignificant difference between the native and the mutant protein. The present study gives astrong indication that the XRCC1 locus deserves serious attention, as it could be a potential candidatecontributing to the etio-pathogenesis of the disease.

Background & objectives: Schizophrenia, the debilitating neuropsychiatric disorder, is known to be heritable, involving complex genetic mechanisms. Several chromosomal regions associated with schizophrenia have been identified during the past; putative gene (s) in question, to be called the global signature for the pathophysiology of the disease, however, seems to evade us. The results obtained from the several population-wise association-non association studies have been diverse. We therefore, undertook the present study on Tamil speaking population in south India to examine the association between the single nucleotide polymorphisms (SNPs) at the serotonin receptor gene (5HT2A) and the occurrence of the disease. Methods: Blood samples collected from 266 cases and 272 controls were subjected to genotyping (PCR amplification of candidate SNPs, RFLP and sequencing). The data on the SNPs were subjected to statistical analysis for assessing the gene frequencies in both the cases and the controls. Results: The study revealed significant association between the genotypic frequencies of the serotonin receptor polymorphism and schizophrenia. SNP analysis revealed that the frequencies of GG (30%, rs6311) and CC genotypes (32%, rs6313), were higher in patients (P<0.05) than in controls. The study also showed presence of G and C alleles in patients. Significant levels of linkage disequilibrium (LD) were found to exist between the genotype frequencies of rs6311 and rs6313. Interpretation & conclusions: This study indicated an association between the SNPs (rs6311 and rs6313) of the serotonin receptor 5HT2A and schizophrenia. HapMap analysis revealed that in its genotype distribution, the Tamil speaking population was different from several other populations across the world, signifying the importance of such ethnicity-based studies to improve our understanding of this complex disease.

Th e investigation was carried out to study the eff ects of Vernonia anthelminticum Willd seed extract on blood glucose level. Th eantihyperglycemic effi cacy of the ethanolic extract of the seed was evaluated in normal, glucose and alloxan induced diabetic rats. The extract exhibited signifi cant hypoglycemic activity in all three animal models when compared with the control group. Th e activity was also comparable to that of the eff ect produced by a standard antidiabetic agent gliclazide, 25 mg/kg (p.o.). Th e results also indicated dose dependent eff ect. Th e hypoglycemia and antihyperglycaemia produced by the extract may be due to increased uptake of glucose at tissue level or increase in pancreatic beta-cell function or due to inhibition of intestinal absorption of glucose. Th e study indicated that the ethanolic extract is a potential antidiabetic agent and lends scientifi c support for its else’s in folk medicine. Key words: Alloxan, antidiabetic, blood glucose, gliclazide, vernonia anthelminiticum