Authors: Alktifan, H1(Undergraduate), Alamri A1(Undergraduate), Alktifan, M1 (Undergraduate), Alneyadi, N1(Undergraduate), Sheryani, M1(Undergraduate), Alyammahi, S1 , Statsenko, Y Dr. 2, Ljubisavljevic, M3, Habuza, T4, Gorkom, K Dr. 2, Zaki, N Prof. 4, Almansoori, T. Dr. 2United Arab Emirates University (UAEU), College of Medicine and Health Sciences (CMHS), Undergraduate MD, Abu Dhabi, United Arab Emirates (UAE). 2 UAEU, CMHS, Department of Radiology, Abu Dhabi, UAE. 3 UAEU, CMHS, Department of Physiology, Abu Dhabi, UAE. 4 UAEU, College of Information Technologies (CIT), Department of Computer Science and Software Engineering, Abu Dhabi, UAE.
Introduction: An assessment of age-related brain structural changes is a common task done by radiologist while reporting brain MRI and CT scans. It is mandatory to highlight the structural signs of brain atrophy leading to cognitive impairment. The evidence coming from the morphological studies of the brain (e.g., voxel-based brain morphometry) may help neurologists to diagnose neurodegenerative conditions at an early stage and contribute to the prophylaxis of the disease worsening.
Aims & Objectives: We intend to study how the brain morphology changes throughout the lifespan in the healthy population. To do so, we collected MRI examinations of the people of different age, then we did the segmentation of the diagnostic images and analyzed the age-related changes of the brain compartment.
Materials and Methods: The study cohort included 231 cases of MRI examinations of people of different ages (4–83 yo). The participants were distributed equally over the following age groups: Adolescents ∈ [0, 20), Young adults ∈ [20, 40), Midlife adults ∈ [40, 60) and Older adults were ≥ 60. The exclusion criteria were as follows: organic brain pathology, mental disorders, injury to the head. To address the research questions, we analyzed the results of brain volumetric studies and built the ordinary least squares regression trendlines for the visualization of the dynamics. To compare the lifelong dynamics of brain atrophy for different brain compartments (e.g., gray matter, white matter, white matter lesions) we prepared the trendlines for each age group and tested the linear models for statistically significant differences between the slopes.
Results: In Young adults and Midlife adults, we did not find significant differences between the rate of the gray matter atrophy, the pace of the white matter atrophy, and the white matter lesions accumulation. The brain intraventricular volume expands to a degree which is significantly lower than the total amount of the cerebrospinal fluid.
Conclusion: The overall atrophy of the gray matter is the most prominent outcome of normal brain aging.
Acknowledgments: This study was supported by SURE+ 2019 Research grants (PIs: KNVG, YS) and StartUp Grant (PI: YS), UAEU.
Authors: 1Osman MH (Graduate student), 1Sulaiman M, 2Hindawi BA, 2Khalil A, 3Galiwango E, 3AlMarzouqi A. and 1Mohsin, S. 1Department of Anatomy, 2Department of Chemistry, 3Department of Chemical and Petroleum Engineering.
Introduction: Bone tissue engineering is a noteworthy technique for bone regeneration to support the injured bones. The ideal bone scaffold should promote early mineralization and support new bone formation while at the same time allow for replacement by new bone. To achieve these goals porous, [11] K.J.L. Burg, S. Porter and J.F. Kellam, Biomaterial developments for bone tissue engineering, Biomaterials 21 (2000) (23), pp. 2347–2359. Article | PDF (541 K) | View Record in Scopus | Cited By in Scopus (272)biocompatible, biodegradable and bioactive material is essential.
Poly(lactide-co-glycolide) (PLGA) has excellent biodegradable properties and are widely used in bone grafting, however, the disadvantage in their use on their own is their poor bioactivity. Around 60 -70% of bone is made of hydroxyapatite. Hydroxyapatite is a biocompatible and bioactive material. We synthesised composite bone scaffolds using nano-hydroxyapatite (n-HAP) in combination with the PLGA. Strontium and zinc were added to the nano-hydroxyapatite to augment bone formation.
Aim and Objectives: To fabricate novel porous composite scaffolds with suitable mechanical properties using zinc (Zn) and strontium (Sr) doped n-HAp with PLGA.
Materials and Methods: Sr/Zn doped n-HAp were synthesized by a water-based sol-gel technique. PLGA was incorporated using the supercritical carbon dioxide technique to form porous composite scaffolds (Sr/Zn n-HAp-PLGA). Samples were mechanically tested under compression with a displacement velocity of 2mm/min using the Universal Testing System Instron 5960 equipped with a 5KN load cell. Specimens were positioned in the center of the loading frame in order to ensure uniform loading and to eliminate moments induced by specimens’ misalignments. The force-displacement response was then converted into a stress-strain response by dividing the force by the cross-sectional surface area and the displacement by the initial length of the sample.
Results: Pure PLGA polymer, n-HAp, and their composites (PLGA-n-HAp) showed ultimate compressive strength ranged between 0.4-19.8 MPa. Doping n-HAp in composite with 2.5% Sr/Zn showed an increase in the ultimate strength by 37% hence the best mechanical properties.
Conclusions: 2.5%Sr/Zn substituted n-HAp-PLGA composite showed a compressive behavior resemble that of cancellous bone indicating that it is a good candidate for cancellous bone tissue engineering.
Acknowledgments: This research was partially carried out using the Core Technology Platforms resources at New York University Abu Dhabi. Research was funded by AARE-2018 and UPAR 2021, G00003460 from UAEU.
Authors: Ali, B., Ahmed, F., Badawi, S., AlKhofash, N. (Graduate); Department of Genetics and Genomics; CMHS; UAEU.
Introduction: COVID-19 caused by SARS-CoV-2 has been declared a pandemic in March 2020. This virus has been proven to be highly infectious and deadly in some cases, causing over 2.5 million deaths world-wide so far and unfortunately, no effective medications are currently available. The entry of the SARS-CoV-2 into human cells to start the infection process is mediated through binding of the viral-spike-protein(S) to a protein receptor, called the angiotensin-converting-enzyme-2 (ACE2), found on the surface of many human cells including lungs. It is therefore predicted that reduced cell-surface availability of ACE2 might reduce the rate of SARS-CoV-2 infection and hence manipulation of cell surface expression of this receptor could present a potent target for COVID-19 treatment. Most ACE2-based therapeutic strategies have aimed to tackle the virus through the use of ACE-inhibitors or neutralizing the virus by exogenous administration of ACE2. Membrane targeting of ACE2, shedding and cellular trafficking pathways, including its internalization, are not well elucidated in literature.
Aims & Objectives: Throughout this study, we aim to 1) elucidate the cellular-trafficking pathways of many identified human ACE2 missense-variants and 2) explore the potency of trafficking inhibitors as COVID-19 therapeutics through their ability to block or slow-down surface expression of ACE2.
Materials & Methods: ACE2 missense-variants are generated by site-directed-mutagenesis using expression vectors as templates. WT and mutant ACE2 trafficking and subcellular localization were then assessed using confocal microscopy and biochemical analysis. Functional analysis of different mutated ACE2 receptors compared to WT-ACE2 is applied to determine their activity and binding capacity to the CoV-2-spike protein via IP analysis. For further understanding at the molecular and structural level; Molecular Dynamics (MD) Simulation to be performed on ACE2 receptor mutants that showed defective binding and retarded trafficking.
Results: To date, 29 missense mutants of ACE2 were generated. WT-ACE2 trafficking, and localization was characterized by immunofluorescence-assay and western-blotting, to be plasma membranal. ACE2 mutants’ characterization, subcellular trafficking and localization compared to WT-ACE2 are being assessed in the meantime.
Conclusions: Our findings might help explain the observed variability in the susceptibility and clinical severity of the disease observed among COVID-19 patients.
Acknowledgments: Project funded by “Abu Dhabi Award for Research Excellence (AARE-2020)”.
Authors: Alktifan H (Undergraduate MD), Alktifan M, Alamri A, Alnuaimi S, Ajab S, Olanda M, Zoughbor S, AlRasbi Z. Department of Microbiology.
Introduction: Blastocystis are intestinal protozoa, showing undetermined pathogenicity and high genetic diversity. According to CDC, 9 subtypes were identified. STs 1, 2, 3 & 4 are the most commonly isolated from human while the rest are thought to be affiliated with zoonosis. Blastocystis infection has been postulated to be associated with chronic disease such as, Irritable Bowel Syndrome, and Colorectal Cancer (CRC). According to a 2011 report released by SEHA, CRC is the second most common cancer and the second highest cause of cancer related deaths in the country. Thus, it is important to investigate gastrointestinal tract cancers. However, a clear causal relationship between chronic diseases and Blastocystis infection has not been investigated thoroughly in the county thus, is yet to be established.
Aim and objective: The aim of the proposed study is to assess the possible association between Blastocystis infection and GTC condition in comparison among cancer patients and healthy population.
Materials & Methods: This study recruits 3 groups of participants: (1) GTC patients, (2) patients with cancers outside gastrointestinal tract (COGT), (3) healthy population. Participants were consented to complete a questionnaire and provide a fresh stool sample. Fecal specimens were analyzed to detect 7 subtypes of Blastocystis sp.: ST1, 2, 3, 4, 5, 6, & 7. Using microscopy, polymerase chain reaction and genotyping.
Results: 53.3% (n=8/15) of cancer patients and 33% (n=5/15) of healthy individuals were positive for at least one Blastocystis subtype by PCR. Blastocystis ST1 was the most common (43.3%, n=13/30). Blastocysitis infection were lower in patient with GTC compared to COGT. Microscopic analysis shows that 20% of healthy individual (n=3/15) and 13.3% of cancer patients (n=2/15) were positive at least for one parasite.
Conclusion: It is hypothesized that GTC patients’ specimens will show higher prevalence of Blastocystis infection compared to specimens of the other two groups. However, the actual findings showed otherwise with the highest Blastocystis prevalence among non-cancer patients then patients with COGT and GTC respectively.
Acknowledgments: This study was supported by SURE+ 2019 Research grant, UAEU.
Authors: Ali N1, Bashir MM1 (Graduate student), Al Maskari F1,2, Elbarazi I1, Oulhaj A1,2, Al-Rifai RH 1,2, Loney T3, Ahmed LA1,2,*. 1 Institute of Public Health, 2 Zayed Centre for Health Sciences, 3 College of Medicine, Mohammed Bin Rashid University of Medicine and Health Sciences, Dubai.
Introduction Gestational diabetes mellitus (GDM) is one of the causes of poor outcomes during pregnancy and later in life, both in mothers and children. Knowledge of GDM burden and its risk factors in a population is paramount for designing targeted preventive interventions in that population. There is currently a dearth of data on the burden of GDM in the Emirati population.
Aims & Objectives To investigate the incidence of GDM in a large representative sample of pregnant Emirati women and identify associated risk factors.
Materials & Methods All pregnant Emirati women aged 18years and above and can provide informed consent, are eligible to participate in The Mutaba’ah Study. This is a multicentre study where data is being obtained through questionnaires and medical data extraction from Al-Ain, Tawam and Kanad hospitals. This index study is an interim analysis of women who joined The Mutaba’ah Study between May 2017 and March 2020. Regression models assessed the relationship between risk factors and GDM.
Results: The analysis included 2,488 women, out of which 675 (27.1%) were diagnosed with GDM in their current pregnancy. For every year increase in maternal age, there is 6% increase in odds of developing GDM (adjusted odds ratio (aOR): 1.06, 95% CI 1.04-1.09). The odds of developing GDM also increased in pregnant women who previously had GDM (aOR: 5.39, 95% CI 4.29-6.77), those with a family history of Diabetes (aOR: 1.41, 95% CI 1.05-1.91), who were pre-pregnancy overweight (aOR: 1.85, 95% CI 1.38-2.50) or obese (aOR: 2.32, 95% CI 1.63-3.29). Interestingly, parity showed an inverse association and seemed protective of GDM.
Conclusion: More than a quarter of women (1 in 4) in the Emirati population developed GDM during their index pregnancies, which is higher than the global burden. Independent risk factors identified include maternal age, pre-pregnancy overweight and obesity, gestational weight gain, family history of diabetes, and history of previous GDM. Focusing on culturally appropriate pre-conception and pregnancy lifestyle interventions may help to reduce the incidence of GDM among Emirati population in the UAE.
Acknowledgements: Funding by grants from Zayed Center for Health Sciences (31R076, 31R183), College of Medicine and Health Sciences (31M266), and the Mohammed Bin Rashid University-Al Mahmeed Collaborative Research Award (ALM1815).
Authors: Jawabri A (Graduate), , Al-Gazali L, Daggag H, Olabi L, Oommen D, Kizhakkedath P, John A, Ali BR Department of Biochemistry, College of Medicine and Health Sciences.
Introduction: Familial hypercholesterolemia (FH) is an autosomal dominant disorder characterized by elevated low density lipoprotein cholesterol (LDL-C) in the blood resulting in coronary artery diseases (CAD) such as atherosclerosis. FH is mainly caused by mutations in a low density lipoprotein receptor (LDLR) gene. The mature form of LDLR resides on the plasma membrane and its role is to capture LDL-C particles circulating in the blood. FH has two forms, the homozygous (HoFH) or heterozygous (HeFH) forms. It has been reported that the prevalence of HeFH is 1:500 while HoFH is 1:1000000 in most populations.
Aims and Objectives: The aim of this research is to establish the cellular pathogenicity of several LDLR missense variants (p. Asp477Asn, p. Gly314Arg, p. Cys243Tyr, p. Asp178Asn, p. Cys167Phe, p. Glu277Lys, p. Arg744Gln, p. His327Tyr, p. Asp622Gly, p. Arg814Gln) identified in Emirati patients by establishing their subcellular localization and glycosylation profiles.
Materials and Methods: The identified missense variants were generated by site directed mutagenesis using Pfu enzyme using the HA tagged LDLR wild type plasmid as template. WT and mutant plasmids were transfected into HeLa and HEK293T cell lines to study their localization using confocal immunofluorescence microscopy and to study their glycosylation profiles by western blot analysis. Calnexin which was used as an ER marker and co-localization with GFP-HRas plasmid used as a plasma membrane marker.
Results: Our results indicate the successful generation of several mutant plasmids and transfection of HA tagged LDLR wildtype plasmid and mutant plasmids into HeLa cells and Hek293T cells. Images obtained by confocal microscopy illustrate that mutants p.Cys167Phe and p. Asp178Asn show complete localization in the ER compared to WT whereas mutant p.Gly314Arg behaved like WT. In addition, the p.Met652Thr showed both cytoplasm and ER localaisations. Western blot analysis confirmed imaging results.
Conclusion: We can conclude that retention in the ER is the most likely underlying mechanism of several LDLR mutations causing FH among Emirati patients.
Acknowledgments: I am grateful to Anne John Mathew for sequencing LDLR mutants and very thankful to Dr. Saeed Tariq for helping with confocal microscopy.
Authors: Alawadhi FMM (Undergraduate), Almarri RHS, Albedwawi AAS, Alraeesi MMS, Hashim MJ, Department of Family Medicine.
Introduction: Hypertension is a major risk factor for cerebrovascular events such stroke as well as hypertensive cardiomyopathy leading to heart failure. The burden of care for these complications is affecting healthcare systems enormously.
Aims & Objectives: We studied the cost-effectiveness of drugs for hypertension.
Materials & Methods: We calculated the mean monthly dosage requirements for commonly prescribed medications used in the treatment of adult patients with hypertension. The typical or mid-range doses were used based on clinical recommendations from Drugs.com. The number of tablets needed per month was obtained based existing formulations of the medication. Next, monthly drug cost was obtained for each medication from the published list of drug prices in the United Arab Emirates. The data source was the United Arab Emirates Ministry of Health and Prevention. We also plotted the price per 10-mmHg drop in systolic blood pressure for each drug to ascertain cost-effectiveness.
Results: A wide range of antihypertensive drugs is available in the UAE. The price per month and per year vary considerably between drug classes and to a lesser extent within a class. For example, enalapril at 10 mg daily costs AED 33 per month; while losartan 10 mg daily for a month is considerably more expensive at AED 88 per month. The difference between generic and brand forms was substantial: nifedipine generic was AED 12 while the brand Adalat was AED 69 per month. Combination pills were not always more expensive: lisinopril (Zestril) 20 mg was priced same as lisinopril with hydrochlorothiazide at AED 90 per month.
Conclusion: The cost of antihypertensive drugs varies considerably in the UAE. Physicians should be aware of these variations (even with insurance coverage) especially as many patients need two or more drugs on a long-term basis.
Authors: Gull B (Graduate), Baby J, Ahmad W, Mustafa F. Department of Biochemistry, CMHS, UAEU.
Introduction: The mouse mammary tumor virus (MMTV) causes breast cancer in mice, leading to tumors that resemble those from human breast cancer. Thus, the MMTV/mouse model is widely used to study human breast cancer. We have previously shown that MMTV dysregulates host microRNAs (miRNA) in MMTV-induced tumors. Since miRNAs are an important component of the RNA interference (RNAi) machinery, this dysregulation suggests a connection between RNAi and MMTV. This is not unique since RNAi plays both pro- and anti-viral roles in different types of viral infections, including hepatitis C, Sindbis, and human immunodeficiency viruses.
Aims & Objectives: Test the hypothesis that “MMTV hijacks the RNAi machinery to facilitate its expression and replication”.
Materials & Methods: An MMTV-expressing stable cell line in HEK293T cells was established using liposomes. miRNA target prediction was performed using RNAhybrid and miRanda software. RNA immunoprecipitations (IP) were performed using the Argonaute antibody. Loss of function analysis was performed by knocking down each gene of the RNAi machinery, including Drosha, Pasha, Dicer, and Argonaute in the MMTV stable cell lines and control cells using shRNA-based vectors. Western blotting was used to confirm knockdowns, while qPCR analysis was conducted to determine effect on MMTV expression.
Results: In silico miRNA target prediction analysis revealed multiple regions in the MMTV genome targeted by several miRNAs dysregulated in MMTV-induced tumors. To confirm whether MMTV was interacting with the RNAi machinery, Argonaute IPs were conducted, demonstrating that MMTV genomic RNA could interact with Argonaute, a key player of the RNA silencing machinery. Knockdown of Drosha led to an increase in MMTV expression, while that of Argonaute led its decrease at both the protein and mRNA levels; however, not much effect was observed upon knockdown of Pasha or Dicer on MMTV expression.
Conclusions: TheRNAi machinery targets the MMTV genome to modulate its expression, with Argonaute facilitating while Drosha inhibiting MMTV expression. The next step is to explore these observations in more detail to determine their significance to MMTV life cycle.
Acknowledgements: This research was supported by a grant (#31R122) from the Zayed Center for Health Sciences, UAEU
Authors: Akhlaq S (Graduate), 2Rizvi TA, 1Mustafa F. 1Department of Biochemistry, 2Department of Microbiology and Immunology, CMHS, UAEU.
Introduction: Viral vectors have been used for gene therapy for decades and recently with improved designs, their use for curing human disorders has become limitless. We have developed efficient and safe vectors for human gene therapy that are based on the mouse mammary tumor virus (MMTV). MMTV has several unique properties that make it a valuable system for the development of effective gene therapy vectors, such as the presence of a tissue-specific, hormone-inducible promoters and most importantly, the ability to infect both dividing and non-dividing cells. This makes MMTV an important viral system for vector development.
Aims & Objectives: Establish a safe yet efficient gene transfer system for use in humans. Specifically, i) construct the next generation of MMTV-based packaging expression vectors and packaging cell lines, ii) construct “self-inactivating (SIN)” gene transfer vectors to deliver the therapeutic gene into target cells, but in the process result in their own “self-inactivation”, iii) test these vectors in vitro in both dividing and non-dividing cells to determine their gene delivery potential.
Results: We have successfully constructed the inducible packaging construct, and verified its expression and transduction potential in comparison to our classical packaging plasmid. A stable packaging cell line has been created using this construct. The packaging plasmid is inducible compared to classical packaging plasmids, allowing us to control its otherwise toxic expression. We have also developed MMTV-based split intron SIN transfer vectors and tested their splicing efficiency and transduction potential to ensure safety. Our split intron SIN vectors containing retroviral splice donor and cloned eukaryotic splice acceptor showed enhanced or reduced splicing efficiency, depending upon the absence or presence of splice sites, respectively. We are also working on the development of MMTV-based deletion SIN transfer vectors that will dual safety in our transfer vectors.
Conclusion: These efforts should lead to the establishment of a new generation of retroviral gene transfer systems that are self-inactivating, yet efficient for gene delivery and their eventual use in human gene transfer studies.
Acknowledgment: This research was supported by a grant (#31R140) from the Zayed Center for Health Sciences, UAEU.
Authors: Alzaabi M (undergraduate), Hraiz N (undergraduate), Bhagavathula A; Al Hamad S, Pathan J, *Aburawi EH.
Introduction: The prevalence of overweight and obesity among children has increased over the past decade. There are little data on the extent of cardio-metabolic risk factors (CMRF) in school-aged children with excess body weight.
Aims & Objectives: To examine the distribution of CMRF across these children through body mass index categories in the UAE to identify the increased CMRF between boys and girls.
Materials & Methods: Cross-sectional survey of the UAE representative sample of children aged 6-17-year-old was conducted in Al Ain in the 2nd, 6th and 10th grades. Multiple logistic regression analysis was performed to investigate the relationship between excess body weight and CMRF between the groups and reported odds ratios (OR).
Results: CMRF such as higher TC, TG, LDL-C, and systolic BP were more prevalent in obese girls. Higher glucose levels, lower HDL-C, and higher diastolic BP were significantly higher among overweight and HbA1c>5g% in obese boys. The OR was 11.6 and 16.9 times among overweight and obese girls for systolic BP. Obese boys and girls were at least 3.5 times more likely to have high TG levels. Overweight boys were 2.6 times more likely to have low HDL-C levels.
Conclusion: Overweight and obesity in school-aged children were associated with increased CMRF, particularly among obese girls aged 12-17 years.Authors: Panicker NG (Graduate), Akhlaq S, Gull B, Baby J, Ahmed W, Mustafa F. Department of Biochemistry, CMHS, UAEU.
Introduction: The mouse mammary tumor virus (MMTV) causes breast cancer in mice via insertional mutagenesis, which allows virus to integrate into mammary epithelial cells, the most permissive cells for virus infection/replication. Activation of steroid hormones during puberty activates MMTV expression, leading to high virus levels, which eventually are passed on to the next generation via the breast milk, perpetuating viral life cycle. Interestingly, MMTV-induced mammary tumors are undifferentiated; yet, mammary gland differentiation is central to MMTV-induced carcinogenesis. High levels of virus replication due to hormonal stimulation during pregnancy and lactation leads to cycles of infection/reinfection of the mammary epithelial cells and multiple random integration events in the vicinity of growth promoting protooncogenes, e.g., Wnt1, Fgf3, and Notch4, activating their expression. This results in clonal expansion of mammary cells carrying MMTV proviruses, and ultimately in tumor formation. Thus, mammary cell differentiation is an important requisite for not only successful spread of MMTV infection between mice, but also tumorigenesis.
Aims & Objectives: Study effect of MMTV expression on host genes during mammary epithelial cell differentiation to determine the cellular pathways affected during virus replication and cell differentiation.
Materials & Methods: HC11 cells stably expressing MMTV were used to study the effects of virus expression on prolactin-induced gene expression. Gene expression profiling utilized mRNAseq, RT-PCRs followed by qRT-PCR.
Results: In vitro differentiation of HC11 mammary epithelial cells was established successfully using multiple independent protocols. MMTV expression during cell differentiation showed inhibition of differentiation as assessed by expression of β-casein gene. mRNAseq analysis revealed a predominant dysregulation of the prolactin-induced pathway of differentiation which was further verified by qPCR. The relevance of these findings to MMTV biology will be discussed.
Conclusions: MMTV alters prolactin-induced gene expression of mammary epithelial cells, suggesting a potential role in mammary carcinogenesis. Given the close similarity between human and mouse breast tumors and the fact that MMTV may be breaking the species barrier, this study provides important insights into the molecular pathways perturbed upon MMTV infection of mammary cells and their transformation.
Acknowledgements: This research was supported by student funds (#31M484) & a CMHS grant (#31M421), UAEU.
Authors: Elsayed N1(Undergraduate), Qureshi MM1, Almansori A1, Bashir G2, Mohamed YA2, AlRamadi BK2, Fernandez-Cabezudo MJ1. Departments of 1Biochemistry and 2Medical Microbiology & Immunology.
Introduction: The cholinergic nervous system innervates multiple organs, including the pancreas. We previously demonstrated that specific inhibition of the enzyme acetylcholinesterase (AChE), which catalyzes the breakdown of the neurotransmitter acetylcholine, prevented the development of autoimmune diabetes in the multiple-low dose streptozotocin (STZ) model. The inhibition of STZ-induced hyperglycemia was achieved by modulating T cell responses and inhibiting pancreatic islet inflammation and β cell loss. Moreover, we have also shown that this cholinergic stimulation induced an increase in mRNA expression of pancreatic insulin. Several studies demonstrated the capacity of pancreatic β cells to self-proliferate by a mechanism that involves multiple cell cycle-related molecules. However, it is not clear yet whether this proliferation is part of β cell regeneration.
Aims & Objectives: To investigate the effect of cholinergic stimulation on the proliferation of pancreatic β cells.
Materials & Methods: BALB/c mice received daily injections of saline, rivastigmine (2 mg/kg) or paraoxon (40nM) for 3 weeks. At the end of treatment, mice were sacrificed, and pancreata excised, enzymatically digested and pancreatic islets isolated. Proteins and/or RNA were extracted from purified islets and analyzed for several markers by western blot and qRT-PCR, respectively.
Results: Paraoxon treatment increased the expression of Reg III (Regenerating islet derived protein III-gamma), Ki67 (proliferation marker) and Foxm1 (forkhead box protein M1) genes. Western blot analysis of β cell extracts from paraoxon and rivastigmine treated mice revealed higher levels of insulin, E2F1(E2 transcription factor 1) and CDK4 (cell division protein kinase 4) proteins than in control saline treated animals.
Conclusions: In addition to increased insulin secretion, our findings demonstrate that cholinergic stimulation also induces proliferation and regeneration of pancreatic β cells. This is evident from the increased expression of proteins involved in β cell regeneration as well as transcription factors that regulate cell-cycle progression.
Acknowledgments: This study is supported by a CMHS grant # 31M427 from UAE University.
Authors: Sultan A1 (Graduate), Adeghate E2, Shafiullah M3, Jacobson M4, Howarth FC1. Departments of 1Physiology, 2Anatomy & 3Pharmacology; 4University of Northwestern, MN, USA.
Introduction: Obesity is a major risk factor for type 2 diabetes mellitus. Cardiovascular disease is the major cause of morbidity and mortality in diabetic patients. Electro-mechanical function is frequently compromised in diabetic heart. Few studies have investigated the effects of obesity and diabesity on the electrical conduction system of the heart.
Aims & Objectives: In vivo biotelemetry techniques were used to evaluate the effects of obesity and diabesity on the electrical conduction system of the heart in the Zucker lean (ZL), Zucker fatty (ZF) and Zucker diabetic fatty (ZDF) rat.
Materials & Methods: Experiments were performed in ZL, ZF and ZDF rats (Charles River, UK). Heart biopotential, physical activity and body temperature were monitored using an in vivo biotelemetry system (Data Sciences, USA). Transmitter devices were surgically implanted in 30 rats (10 ZL, 10 ZF, 10 ZDF) aged 75 days under general anesthesia. The devices were inserted into the peritoneal cavity and the electrodes were arranged in Einthoven Lead II configuration (right foreleg and left hindleg). Ethical approval was obtained from the Animal Research Ethics Committee, UAE University. Two-way ANOVA and Independent Samples t Test was used for statistical analysis.
Results: At 6.5 months of age, heart rates (HR) were significantly (p<0.05) lower in ZDF (265±8 bpm, n=10) compared to ZF (336±9 bpm, n=10) and ZL (336±10 bpm, n=10) and heart rate variability (HRV) showed significant differences between ZDF (22±1 bpm, n=10), ZF (27±1 bpm, n=10) and ZL (31±1 bpm, n=10) rats. The electrocardiogram showed an age-dependent prolongation of PQ and QRS intervals in all 3 groups of rats and the QRS interval was significantly prolonged in ZDF compared to ZF rats. Power spectral analysis revealed no significant (p>0.05) differences in HRV at low frequencies, reduced HRV at high frequencies and increased sympathovagal balance in ZDF compared to ZF and ZL rats.
Conclusion: HR was reduced by aging and additionally reduced by diabesity in the absence of changes in physical activity and body temperature. Reductions in HRV associated with altered sympathovagal drive might partly underlie disturbed HR in the ZDF rat.
Acknowledgments: Zayed Center for Health Sciences, UAEU, Grant 31R133
Authors: Samha HI2 (Graduate), Ali BR1,2,3. 1 Department of Pathology, College of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, United Arab Emirates.2 Department of Genetics and Genomics, College of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, United Arab Emirates. 3 Zayed Centre for Health sciences, United Arab Emirates University, Al-Ain, United Arab Emirates.
Introduction: Low-density lipoprotein receptor related protein 6 (LRP6) is a member of the low-density lipoprotein receptors family (LDLRs) which is involved in the clearance of LDL from the blood stream and the signaling of the canonical wingless signaling pathway. Several genetic variants in LRP6 gene have been conclusively associated with coronary artery diseases (CADs). However, the structural and functional implications of those variations have not been fully elucidated. Previous studies revealed that LRP6 mutations associated with Oligodontia disease are found retained in the endoplasmic reticulum (ER) and therefore not expressed on the cell surface. The CAD-associated LRP6 variants could be similarly retained and possibly subjected to ER associated degradation (ERAD); an adaptive process employed by cells to eliminate misfolded proteins to maintain ER homeostasis.
Aims & Objectives: To investigate the sub-cellular localisations, stability, and transport of ten LRP6 missense mutants associated with CAD.
Materials & Methods: Ten LRP6 missense CAD-associated variants were generated via site directed mutagenesis. Mutated protein molecular processing and localisation were assayed via immunofluorescence and western blot. HeLa cells were transfected with wild-type and mutated plasmids to access LRP6 co-localisation with ER and plasma membrane markers. For protein processing and glycosylation profiles analysis, the wild-type and mutant plasmids were transfected into HEK293T, and then the protein lysates were used for Endoglycosidase H sensitivity assay and western blotting.
Results: Our findings indicate that two of the CAD-associated LRP6 variants (p.Tyr418His and p.Asn433Ser) were ER retained. These mutations were co-localised with calnexin (ER marker) as indicated via immunofluorescence analysis. Protein processing of these two variants to their plasma membrane mature form were reduced as indicated via western blot confirming the previously obtained IF results. The glycosylation profile for the wild-type and the mutated LRP6 were tested via Endoglycosidase-H digestion to further study the molecular effect of the underlying genetic variants.
Conclusions: ER retention of CAD-associated LRP6 variants could contribute to their pathogenicity. Further work will include functional assays and test their association with ERAD components. This study will lead to better understanding of the pathogenesis of LRP6 mutations with potential applications in clinical diagnosis and development of new therapiesAuthors: Al-Saafeen BH1 (Graduate), Mohamed YA1, Bashir G1, Haneefa SM1, Alsbiei A2, Fernandez-Cabezudo MJ2, and Al-Ramadi BK1. Departments of 1Medical Microbiology & Immunology, and 2Biochemistry, College of Medicine & Health Sciences, United Arab Emirates University, United Arab Emirates.
Introduction: Cancer is a major concern worldwide with nearly 9.6 million deaths each year. It is now recognized that cancer development and progression is linked to a compromised immune system. The development of immune checkpoint inhibitors has led to unprecedented clinical benefits for cancer patients in recent years. However, the response rate among patients remains disappointingly low. Previously, we demonstrated the potential of using attenuated bacteria as an anti-cancer and immunomodulatory agent. In the current study, we have investigated the potential of utilizing a low dose of attenuated bacteria to enhance the outcome to treatment with checkpoint inhibitors.
Aims & Objectives: To investigate the effect of a low dose of attenuated Salmonella typhimurium on tumor growth and to study its immunomodulatory effect using MC38 murine colon adenocarcinoma model.
Materials & Methods: C57BL/6 were inoculated s.c. with MC38 cells and staged to day 7 when visible tumors began to be observed. Tumor-bearing mice were then inoculated i.p. with either saline or attenuated Salmonella typhimurium (~5× 103 CFUs/mouse). Tumor volumes were measured twice a week. Mice were sacrificed two weeks following bacterial treatment. Tumor and spleen tissues were collected for further analysis. Multi-color Flowcytometry and immunohistochemistry were used to study bacterial-induced changes in anti-tumor immune responses.
Results: A low dose of attenuated Salmonella resulted in ~40% reduction in MC38 tumor growth accompanied with a pronounced splenomegaly. Flowcytometric analysis revealed that splenomegaly is mediated by infiltration of both lymphoid and myeloid immune cells. At the tumor level, our results also revealed Salmonella’s ability to (1) enhance the infiltration of CD4+ helper and CD8+ cytotoxic T cells into the tumor and (2) alter the expression of checkpoint inhibitors on immune cells.
Conclusion: Our findings highlight the capacity of a low dose of Salmonella to create a more conducive tumor microenvironment for improving the outcome to checkpoint inhibitors.
Acknowledgments: This study is supported by grants from UAEU and the Terry Fox International Research Foundation.
Authors: Gariballa N (Graduate), Kizhakkedath P, Ali BR. Department of Genetics and Genomics.
Introduction: Hereditary hemorrhagic telangiectasia type 1 (HHT1), also known as Rendu-Osler-Weber syndrome, is an autosomaly dominant inherited disease that is generally characterized by vascular malformation and fragility. HHT1 has been associated with mutations in the TGFb co-receptor Endoglin, encoded by ENG gene. We have previously demonstrated that some Endoglin variants are trapped in the Endoplasmic Reticulum (ER) and fail to traffic to their normal localization in plasma membrane, which suggested the involvement of ER associated protein degradation (ERAD) in their molecular pathology.
Aims & Objectives: Investigate the degradation pathway of wild type (WT) and mutant Endoglin retained in the ER in order in elucidate the exact molecular mechanism underlying the loss of function phenotype associated with this disease. Furthermore, we aim to generate HEK-293 cell lines deficient in major ERAD components (e.g. HRD1 E3 ubiquitin ligase) using CRISPR cas9 gene editing technology. Stability of WT and mutant Endoglin variants in these cell lines will then be determined.
Materials & methods: Stable HEK293 cell lines harboring the HA-tagged ENG-WT and mutant variants P165L and V105D were generated. Cycloheximide chase assay was used to determine the cellular half-life of ENG-WT and mutant variants. To delineate the degradation pathways, HEK293 cells were incubated with ERAD, proteasomal, or lysosomal inhibitors (Eeyarestatin I, and Kifunesine /Bafilomycin, chloroquine/ MG132, Epoxomycin, respectively). Western blot was performed to measure protein accumulation level. HEK293(HRD1-KO) were generated using (Origene) HRD1 KN.2 kit for CRISPR Cas9 gene Knock out (KO).
Results: Our data shows that WT Endoglin is degraded through both proteasomal and lysosomal pathways, while the mutant variants, trapped in the ER, undergo proteasomal degradation. Furthermore, preliminary results illustrated the accumulation of both WT Endoglin and mutant variants (P165L&V105D) when transiently transfected into HEK293(HRD1-KO) cell line compared to control cell line suggesting their stabilization.
Conclusion: In this study, we demonstrate for the first time the implication of ERAD in the pathogenesis of HHT1 disease through the elucidation of the degradation pathways of both wild type (WT) and disease-causing mutant variants of Endoglin.
Acknowledgements: We thank UAE University and ADEK for funding our project through (AARE-19-086) and UAEU (31M439) grants, respectively.
Authors: Challagandla AK (Graduate), Kaimala S, Daniel AP, Sreedharan SP, Emerald BS. Department of Anatomy, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, Abu Dhabi, UAE.
Introduction: MicroRNAs (miRNAs) are ~21bp long regulatory RNAs and involves in the regulation of cellular pathways by targeting associated mRNAs. They regulate all known cellular functions such as development, differentiation and metabolism and dysregulation in their expression have been linked to different diseases including metabolic diseases such as obesity and type 2 diabetes. Even though rat is a well-established model system for metabolic studies and the relevance of miRNAs as regulators of metabolism is well established, very few miRNAs have been identified in rat, compared to humans and mouse. We carried out a genome-wide small RNA sequencing using rat skeletal muscle and compared two developmental stages, newborn (day 1) and adult (day 145) rats and two developmental conditions, lower birth weight (LBW, 5th to 25th percentile) and average birth weight (ABW, 50th to 75th percentile). Further small RNA_Seq analysis and miRNA functional verification was carried to investigate the identified novel miRNAs involved in metabolic pathways and associated complications.
Aims & Objectives: Identifythe small RNA transcriptome repertoire of developmental and metabolic significance using a rat model system.
Materials & Methods: Whole-genome small RNA-sequencing was carried to identify the small RNA transcriptome repertoire involved in metabolic pathways. L6 muscle satellite cell line was used for in-vitro miRNA knockdown/overexpression studies and functional characterization of the identified miRNAs.
Results: We identified a total of 2932 miRNAs of which 1976 miRNAs were previously described at least in one species (human, mice or rat) and 956 previously undescribed small-RNAs whose expression changed during development or altered as a function of their developmental condition. We have verified the developmental expression of 103 of these novel pre-miRNAs. Further, by extending the analyses to 23 of these mature miRNAs, we have identified the gene targets and the pathways they may regulate. We also show the significance of two of these novel miRNAs to lipid metabolism and obesity.
Discussion: We have identified 956 novel small-RNAs. As these are of either developmental or metabolic relevance these will be a good resource for further analyses which will contribute significantly to our understanding of miRNA biology in development and disease.
Acknowledgement: This work is supported by UPAR and Zayed Centre based grants from UAE University.
Authors: Elsayed N (undergraduate)2; Peck G1; Hashim2; Shaughnessy C1; Muddasani S1, Fleischer A1. 1Department of Dermatology, University of Cincinnati, USA; 2College of Medicine and Health Sciences, UAE University, Al Ain.
Introduction: Urticaria is a common condition that presents with transient, pruritic wheals, angioedema, or both. It has a high socioeconomic burden worldwide. Currently, the global epidemiology of urticaria and its geographical and temporal trends are not well studied in existing literature. By characterizing the global patterns of disease, clinicians and public health professionals will be able to better understand and reduce the impact of urticaria.
Aims & Objectives: This study aimed to evaluate the global patterns and trends for urticaria over the last three decades, and to assess the effect of socioeconomic development on urticaria prevalence.
Materials & Methods: Using the Global Burden of Disease (GBD) dataset, we analyzed the age-standardized prevalence, incidence, and years lived with disability (YLD) of urticaria in 195 countries from 1990 to 2017. Additionally, the relationship between gross-domestic product (GDP) and the disease burden of urticaria was evaluated.
Results: The global prevalence of urticaria was 86 million people in 2017, roughly 1.1% of the global population. Females and children aged one to four were more commonly affected—these differences were outside the 95% uncertainty intervals. The global incidence rate was 2,512 per 100,000 among 1- to 4-year-old children. The age-standardized global prevalence rate remained stable from 1,119 cases per 100,000 in 1990, to 1,126 per 100,000 in 2017. Regression analyses showed that a higher GDP per capita was positively associated with prevalence and incidence (p < 0.001).
Conclusions: Urticaria is more common in females, children aged one to four, and regions with lower GDP per capita. Global prevalence, incidence, and YLDs have remained stable between 1990 and 2017.
Acknowledgments: I would like to acknowledge my supervisor Dr. M Jawad Hashim and thank all the participants and all those who helped us completing this research.
Authors: Baby J (Graduate), Gull B, Ahmed W, Mustafa F. Department of Biochemistry, CMHS,
UAEU.
Introduction: The mouse mammary tumor virus (MMTV) is well recognized for its ability to develop
breast cancer in mice. Not much is known about how MMTV interacts with host microRNAs
(miRNAs), well-known post-transcriptional regulators of gene expression that modulate
key cellular functions. These non-coding RNAs are also potent regulators of viral
infection, replication and pathogenesis. We have previously demonstrated that MMTV
does not encode for its own miRNAs, but dysregulates host miRNAs, including members
of the oncogenic miR-17-92 cluster.
Aims & Objectives: Characterize the effect of MMTV on the expression of miR-17-92 cluster in mammary epithelial and non-mammary cells. Conduct functional analysis of over-expression/inhibition of these miRNAs and identify downstream target genes that modulate MMTV expression and replication.
Materials & Methods: miRNAseq andmiRNome qPCR profiling were used to confirm dysregulation of miR-17-92 in MMTV-expressing stable cell lines (HC11 and HEK293T). Lipofectamine-based transfections were used to establish dual stable cell lines either over-expressing or inhibiting individual miRNAs in the MMTV stables to conduct functional assays. Effect of miRNA over-expression/inhibition on MMTV expression was assessed by Taqman qPCR assays. Target gene analysis of specific miRNAs was conducted by western blots.
Results: miRNAseq and miRnome analyses confirmed dysregulation of several host miRNAs, including miR-17-92 cluster members in cells expressing MMTV. Interestingly, inhibition of miRNAs using specialized PMIS vectors showed an upregulation of these miRNAs by qPCRs rather than their downregulation despite upregulation of their target PTEN. We hypothesize that this phenomenon could be partially due to segregation of inhibited miRNAs in P-bodies, sites of RNA-protein (RNP) complexes that sequester translationally-repressed mRNAs and proteins related to mRNA decay. The downregulation of miR-17,18,19 resulted in up-regulation of MMTV in stable cell lines. Conversely, over-expressing of individual cluster members (miR-17,18,19 and 92) was seen to downregulate MMTV expression.
Conclusions: MMTV expression dysregulates the expression of miR-17-92 cluster, a critical class of miRNAs that act as oncogenes, suggesting that MMTV infection predisposes the mammary epithelial cells to cell transformation and eventual cancer.
Acknowledgments: This research was supported by a grant (#31R122) from the Zayed Center for Health Sciences, UAEU.
Authors: Bhagavathula AS (Graduate), Shah SM, Aburawi EH, Oulhaj A (Supervisor). Institute of Public Health.
Introduction: Few studies have investigated the achievement of blood pressure goals following these guidelines in patients with cardiometabolic risk factors.
Aim & Objectives: To evaluate the change in BP in six months of treatment initiation and compare the achievement of BP goals according to the American (ACC/AHA, 2017, European (ESC/ESH, 2018), United Kingdom (NICE, 2019), and the International Society Hypertension (ISH, 2020) guidelines. We also investigated the factors associated with guidelines recommended BP goals according to their cardiometabolic status.
Methods: We analyzed data of 5308 newly diagnosed hypertensive outpatients in Abu Dhabi, UAE, in 2017. Hypertension was defined as BP 130/80 mmHg according to the ACC/AHA and 140/90 mmHg following ESC/ESH, NICE, and ISH guidelines. Mean changes in the BP during the six months follow-up was evaluated. Multiple logistic regression was performed to identify the factors associated with an increase in the achievement of guidelines-recommended BP goals.
Results: Overall, the mean BP was 133.9 ± 72.9 mmHg at baseline and 132.7 ± 72.5 mmHg at six months. Guidelines-recommended BP goal achievement was 45.5% according to the ACC/AHA, the ESC/ESH (75.3%), the NICE (66.3%), and the ISH was 72.2%, respectively. The hypertension control among patients with and without cardiometabolic risk factors at 6-months was 64.7% and 59.1%. Aspirin therapy (odds ratio (OR): 4.09; 95% confidence interval (CI), 1.76–9.48) was associated with the achievement of the ACC/AHA recommended BP goals. Young age (OR = 2.66; 95% CI: 1.10–6.40) was associated with the ESC/ESH BP goals, and antihypertensive medication adherence was associated with BP goals according to the NICE (OR = 3.09; 95% CI: 1.90–5.04) and the ISH (OR= 2.72, 95% CI: 1.59–4.65) guidelines
Conclusion: BP goal achievement was suboptimal in hypertensives with/without cardiometabolic risk factors in the UAE. Patients of young age, aspirin therapy, and antihypertensive medication adherence are significant predictors to achieve guidelines-recommended BP targets. BP control efforts should prioritize improvement in medication adherence, cardiometabolic goals, and lifestyle.
Acknowledgement: This work is supported by the educational grant no. 21M092.
Authors: Al-Azawi A.1 (Graduate), Arafat K.1, Sulaiman S.1, Attoub S.1Department of Pharmacology and Therapeutics, College of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, UAE.
Introduction: Lung cancer is one of the most common forms of cancer with the highest mortality rate in the world accounting for 1.76 million deaths in 2018 and this despite the advances in targeted- and immuno-therapies. DCA targeting cancer cell maetabolism could be a promising strategy for the treatment of cancer.
Aims & Objectives: Whether DCA alone or in combination with the EGFR targeted therapies “Gefitinib and Erlotinib” is a successful approach for lung cancer therapy was our major objective.
Materials & Methods: This was addressed using two lung cancer cells namely A549 and LNM35 through the investigation of the impact of DCA alone and in combination with the proposed therapies on lung cancer cell viability, migration, invasion, and colony formation in vitro and on tumor growth and metastasis using the chick embryo and the nude mice models in vivo. The antiangiogenic potential of DCA and its safety profile were also investigated.
Results: We demonstrate that DCA causes a concentration- and time-dependent decrease in the viability of A549 and LNM35 cells and the growth of their colonies in vitro. Similarly, DCA slowdown the growth of A549 and LNM35 tumor xenografts in both the chick embryo and nude mice models in vivo. DCA decrease the angiogenic capacity of human umbilical vein endothelial cells (HUVECs), suggesting the inhibition of tumor angiogenesis as a potential mechanism behind its anti-tumor growth effect. On the other hand, DCA didn’t decrease the growth of lymph nodes metastasis in nude mice xenografted with the highly metastatic lung cancer cells LNM35. Treatment with DCA did not lead to significant side effects on the embryo viability or on the nude mice weight. In addition, blood, kidney, and liver function tests showed no toxicity with DCA when compared to the control group. Finally, we demonstrate that DCA significantly enhanced the inhibitory effect of gefitinib on the cellular viability and clonogenic growth of A549 and LNM35 cells.
Conclusion: This study paves the way for future pre-clinical investigation of the potential benefit of combining DCA with the currently used lung cancer targeted therapies.
Authors: Al Hosani M (undergraduate), Shouk R, Karam SM, Department of Anatomy.
Introduction: The uterine cervix is composed of two regions: ectocervix and endocervix. While the ectocervix is covered by stratified squamous epithelium, the endocervix is lined by simple columnar epithelium. This junctional epithelium is a common site for pathological changes. Lectins are naturally occurring proteins with affinity to specific carbohydrate moieties, therefore, can be used as markers for different cells in these regions.
Aims & Objectives: This study aims to characterize the binding affinity of various lectins to the cervical epithelium with a special focus on the differences in the junctional area during the four stages of estrus cycle.
Materials & Methods: Female virgin C57BL/6 mice (n=16) at the age of 2-4 months were used. To identify stages of estrus cycle, mice were evaluated cytologically via vaginal lavage. Cervical tissues were processed for histological examination to confirm the four stages of estrus cycle. Then, a panel of lectins were finally used to analyze their binding affinity to different epithelial cells of the uterine cervix.
Results: Mice in the four stages of estrus cycle were identified using cytological and histochemical methods. In proestrus stage, most cells in vaginal smears were round nucleated epithelial cells. Histologically, mucified and keratinized epithelial layers began to appear. In estrus stage, smears showed cornified squamous epithelial cells and the keratin layer became superficial and thick. In metestrus stage, a mix of nucleated epithelial and/or cornified squamous epithelial cells with many leukocytes dominated in the smears. Histologically, the epithelium was infiltrated with leukocytes. Diestrus stage showed smears with predominance of leukocytes. However, the apical epithelial layer was mucified with luminal mucus, leukocytes and desquamated cells. The binding affinity of 20 different lectins to the epithelial lining of ectocervix and endocervix was characterized for each stage of estrous cycle. Lectins specific for galactose, N-acetyl-glucosamine, fucose, and mannose/glucose differentially bound along the cervical epithelium and the pattern varied with the different stages.
Conclusions: Characterization of different stages of estrus cycle and the lectin binding pattern to cervical epithelial cells could provide the basis for better understanding of the pathogenesis of some common disorders that affect the uterine cervix including tumors.
Acknowledgment: This research was supported by SURE grant and College of Graduate Studies, UAEU.
Authors: Lootah S,(Graduate), Kaimala S, Sreejisha PS, Princy J, Emerald BS. Department of Anatomy, CHMS, UAE University.
Introduction: Long non-coding RNAs (lncRNAs) exert an additional layer of gene regulation. However, the biological relevance of few lncRNAs are documented. Insulin (INS) is an important hormone with pivotal roles in glucose homeostasis. Its secretion is tightly regulated with nutrients and other insulin secretagogues eliciting a range of cellular signals in b-cells resulting in Insulin gene transcription and secreation. Several transcription factors such as MAFA, NKX2.2, NKX6.1, PDX1 regulate INS expression. PDX1 is the master regulator of b-cell development and function. We have identified a lncRNA, UAE-710, which is transcribed in an anti-sense orientation to PDX1.
Aims & Objectives: To systematically characterize the lncRNA UAE-710, and to establish the possible regulatory role in INS expression. We are also interested in establishing the regulatory pathway through which lncRNA-UAE710 mediates its regulatory role.
Materials & Methods: We have cloned the lncRNA UAE-710 by using standard molecular biology techniques and established human insulin-secreting b-cell line 1.1B4 with over-expression of lncRNA UAE-710 and also generated 1.1B4 knocked-out cells (UAE-710 D- 1.1B4) using CRISPR-Cas9 technology. Using these cell lines, we have verified the expression of INS and PDX1 by qRT-PCR and western-blot. We have also investigated the mechanism of action of UAE-710 in 1.1B4 cells by RNA-centric strategies, Chromatin Immunoprecipitation by RNA Precipitation (CHIRP), and RNA immunoprecipitation (RIP). We also performed glucose stimulation experiments to verify the response of lncRNA UAE-710 to glucose challenge.
We also verified the transcript by northern blotting in 1.1B4 and confirmed its expression in different adult and human fetal tissues by qRT-PCR.
Results: Human pancreatic b-cells, as well as other human adult and fetal tissues express lncRNA UAE-710 at different levels. A high glucoseenvironment upregulates lncRNA UAE-710 expression in 1.1B4 cells. Its overexpressionresults in PDX1 independent upregulation of INS expression. Similarly lncRNA UAE-710 D-1.1B4 cells showed a significant reduction in INS. CHIRP experiment had demonstrated that UAE-710bind to INS promoter and as well as coding sequences.
Conclusion: LncRNA UAE-710 is a positive regulator of INS regulation in human pancreatic b-cells. However, its role in INS secretion and regulation is independent of PDX1, probably by binding to the INS promoter.
Acknowledgment: This work is supported by Zayed center: 31R228.
Authors: Masad RJ1 (Graduate), Mohamed YA1, Bashir G1, Amer R1, Alsbiei A2, Fernandez-Cabezudo MJ2, and Al-Ramadi BK1. Departments of 1Medical Microbiology & Immunology, and 2Biochemistry, College of Medicine & Health Sciences, United Arab Emirates University, United Arab Emirates.
Introduction: The focus on using alternative natural products for cancer prevention and treatment has been increasing over the past few years. Among these natural products, Manuka honey (MH) has been extensively researched. We previously demonstrated that MH can inhibit the growth of several human cancer cells in vitro, including melanoma, breast and colorectal cancer. Furthermore, using an in vivo syngeneic mouse melanoma model, intravenous administration of MH enhanced the therapeutic efficacy of paclitaxel and improved overall host survival.
Aims & Objectives: In the present study, we aimed to investigate the effect of oral administration of MH on tumor growth using a syngeneic model of colorectal cancer, and to identify the mechanism by which MH can influence the anti-tumor immune response.
Materials & Methods: MH was administered daily by oral gavage starting 2 weeks prior to tumor implantation and continued for two more weeks after implantation. Control mice were either left untreated or given a mixture of the three main sugars found in MH. Tumor growth was followed for the subsequent weeks. Mice were sacrificed 3-4 weeks post implantation, and tumors and spleens were excised for further analysis.
Results: Tumor growth was significantly inhibited (p<0.05) in MH-treated group compared to the control groups. This was accompanied by a significant decrease in the degree of splenomegaly among MH-treated mice. Flowcytometric analysis revealed a significant decrease in the accumulation of myeloid cells in the spleens of MH-treated mice, and an increase in the percentage and activation of tumor infiltrating cytotoxic T cells. Immunohistochemical analysis of tumor tissues showed an increase in the number of cytotoxic T cells and GZM-B-expressing cells following MH-treatment. In addition, analysis of gene expression in purified tumor-infiltrating leucocytes highlighted an enhancement in expression of proinflammatory cytokines and a decrease in the expression of immunosuppressive cytokines following MH treatment.
Conclusion: Our results suggest that in addition to its demonstrated tumoricidal potential, MH may adversely influence tumor growth by boosting anti-cancer immune response. These findings highlight a potential role for MH in the prevention of colon cancer.
Acknowledgments: This study is supported by a UPAR grant #G00002993 from UAEU.
Authors: Alawathugoda TT (Graduate), Ardah MT, Ansari SA. Department of Biochemistry.
Introduction: The neurodevelopmental disorders have a complex etiology that might be similar and start very early-on during embryonic development through the involvement of epigenetic mechanisms. Previous research based on this, in our lab, found that saturated fatty acids alter developmental neurogenesis due to reduced progenitor proliferation and increased differentiation into neurons. It was also found that the expression of SREBPs, the transcription factors responsible for de novo fatty acid and cholesterol biosynthesis were significantly reduced due to fat treatment at a specific stage of neurogenesis. The expression of SREBPs are regulated by SIRT1 and LSD1 which are histone deacetylase and demethylase respectively, which in turn also regulate Notch signaling pathway, an important regulator of developmental neurogenesis. Thus, it could be hypothesized that SIRT1-LSD1 acting as lipid sensor could alter SREBP dependent lipid metabolism due to excess fat which could then affect Notch pathway leading to defective neurogenesis.
Aims & Objectives: 1. Genome wide high-throughput RNA-sequencing to identify the genes differently expressed due to high saturated fatty acid levels in the course of human embryonic neurogenesis. 2. The effect of saturated fat on global histone acetylation and methylation by high-throughput ChIP-Seq during cortical neurogenesis. 3. Identify the role of SIRT1-LSD1 on SREBP and Notch signaling pathway during embryonic neurogenesis.
4. Functional validation of genome wide identified mRNA targets and promoter sequences by mRNA-Seq and ChIP-Seq data. 5. Validation of key results from human stem cell model in an animal model of high fat diet induced obesity.
Materials & Methods: Human embryonic stem cells (hESCs) that have the capacity to be differentiated into any cell type of the body, will be used to understand the relationship between increased levels of saturated fatty acids such as palmitate on embryonic neurogenesis in human. They will be subjected to directed differentiation into cortical neurons as a model of early human embryonic neurodevelopment. Using this model, we will study the effect of saturated fatty acid on mRNA expression and histone modification during embryonic neurogenesis.
Results & Conclusion: The identification and functional validation of novel molecular mechanism/s of saturated fatty acid mediated alterations on embryonic neurodevelopment at the transcriptional/epigenetic level, will realized using advanced techniques in stem cell culture and differentiation, high-throughput RNA and PCR techniques, chromatin immunoprecipitation-sequencing (ChIP-Seq), immunocytochemistry and an array of molecular and biochemical techniques. The knowledge thus gained will be critical in the development of preventative strategies and therapeutic interventions for Neurodevelopmental Disorders.
Authors: Amer R1 (Graduate), Bashir G1, Alsbiei A2, Altahrawi A3, Fernandez-Cabezudo MJ2, and Al-Ramadi BK1. Departments of 1Medical Microbiology & Immunology, 2Biochemistry, and 3Pathology, CMHS, UAE University.
Introduction: The activation of the endothelin receptor type A (ETAR) by its ligand endothelin-1 (ET-1) is known to induce vasoconstriction. The ETAR antagonist, Ambrisentan, is FDA-approved for the treatment of pulmonary arterial hypertension. Interestingly, tumor cells also express ETAR and ET-1. Activation of ET-1 axis promotes cell proliferation, survival, and epithelial-to-mesenchymal transition.
Aims & Objectives: To investigate the effect of Ambrisentan, a selective antagonist of ETAR, on tumor growth and metastasis using a syngeneic, orthotopic triple negative breast cancer (4T1) animal model.
Materials & Methods: BALB/c mice were divided into control and Ambrisentan group (n= 8-10 mice/group). Ambrisentan (10mg/kg) was administered by daily oral gavage for 3 weeks before, and another 2 weeks post, tumor implantation. 4T1 or 4T1-Luc2 (luciferase-transfectant) cells were inoculated s.c. in the mammary fat pad. Tumor growth and animal survival were followed for up to 6 weeks. In some studies, mice were sacrificed 3 weeks post implantation and their tumor and organs (spleen, lung and liver) were processed for analysis to assess the extent of tumor metastasis.
Results: Treatment with Ambrisentan significantly reduced tumor growth and improved overall survival of tumor-bearing mice. This was correlated with 40-60% reduction in tumor metastasis to major organs, including liver and lungs. Using 4T1-Luc2 tumor cells and in vivo life imaging (IVIS) of animals revealed 5- and 18-fold decrease in the bioluminescence signal collected from the primary tumor site and distant organs, respectively, in Ambrisentan treated mice. This was further confirmed by quantifying the actual number of tumor cells metastasizing to the lungs using 4T1-Luc2 cells. To understand the underlying mechanism for the anti-metastatic effect of Ambrisentan, we examined the extent of angiogenesis within the tumor tissue. The findings revealed that total tumor vascularity was reduced by ~50%, mainly due to a decrease in the size of blood vessels in animals treated with Ambrisentan.
Conclusion: Ourfindings demonstrate a significant and beneficial effect of Ambrisentan in this highly metastatic breast cancer model and provide a rationale for targeting ETAR as a potential adjuvant therapy in cancer patients.
Acknowledgments: This study is supported by grant #AJF201709 from Al Jalila Foundation
Authors: Prabakaran AD (Graduate), Kaimala S, Sreedharan SP, Vijayan R, Emerald BS. Department of Anatomy.
Introduction: The molecular mechanism/s which govern the metabolism and changes leading to the increased risk of developing obesity is still not clear. From a microarray analysis aimed at identifying genes which respond to shuttle changes in the nutrition during early embryonic development we have identified clustered Protocadherins as one set of possible regulatory molecules. Herein, we verify the role of PCDHa4 mediated signaling mechanism in modulating lipid metabolism and how changes in them results in obesity using a rodent model system.
Aims & Objectives: The objective of this study was to utilize a rodent model system and to identify the molecular mechanism/s mediating the altered susceptibility towards metabolic syndrome including obesity.
Materials & Methods: Using birth weight as a marker and two groups of the normal birth weight range of rat pups, lower birth weight and average birth weight we have carried out a microarray analysis and established the associated pathways. After the verification of the array by quantitative PCR, muscle and liver samples were used for the analysis of glucose and fat metabolism. High Fat Diet (HFD) adults were also used to further verify the metabolic and molecular changes related to obesity. The role of PCDHa4 and the associated regulatory pathways were analyzed using appropriate techniques such as MeDIP-qPCR, ChIP-qPCR, EMSA, Western blot and Co-IP.
Results: Microarray analysis showed the expression of Protocadherins were altered in LBW adults. Methylation analysis using MeDIP-qPCR showed increased methylation in the regulatory region of PCDHa4 which correlated with the decreased expression of PCDHa4 and correspondingly affected the binding of transcription factor E2F3 as evidenced by ChIP-qPCR and EMSA analysis. Further, Co-IP analysis showed formation of a stable functional complex involving RET-GDNF-GDNFR-alpha which regulates the genes in lipid metabolism including, SREBP1, PPARγ, C/EBPb, C/EBPa. This was altered in LBW and HFD adults. Together our analysis establishes a role for PCDHa cluster in obesity.
Discussion: Our results suggest a link between PCDHa cluster mediated functional complex in maintaining lipid metabolism and changes in this may have a role in obesity.
Acknowledgement: This work is supported by UPAR and Zayed Centre based grants from UAE University.
Authors: 1Khader TA (Graduate), 1Ahmed W, 1Akhlaq S, 2Rizvi TA, 1 Mustafa F. 1Department of Biochemistry, 2Department of Microbiology and Immunology, CMHS, UAEU.
Introduction: Mouse mammary tumor virus (MMTV) causes breast cancer in mice. During replication, MMTV must export its genomic RNA (gRNA) to the cytoplasm without splicing. This is ensured by the signal peptide of Rem that interacts with a 3’ cis-acting RNA element, the Rem-responsive element (RmRE), to facilitate the nucleocytoplasmic transport of the gRNA. Furthermore, we have identified a 5’ cis-acting element that facilitates the transcription, stability, and elongation of the gRNA. However, whether this element is Rem-responsive or has any functional interaction with the 3’RmRE to facilitate MMTV gene expression is not clear since all assays were conducted in the presence of the 3’RmRE.
Aims & Objectives: Investigate role of the 5’ element in transcriptional regulation of MMTV gRNA independent of the 3’RmRE. Hypothesis: Either a transcriptional elongation factor and/or RNA stability protein binds to this region to enhance expression of full-length gRNA, while Rem/RmRE ensures its safe export to the cytoplasm.
Materials & Methods: The 5’ element and 3’RmRE were deleted either alone or together in a molecular clone of MMTV. Stable HEK293T cells expressing mutant clones were generated and treated with Actinomycin D to inhibit transcription with time to study transcription, stability, elongation, and transport of the gRNA. qRT-PCRs were used on cytoplasmic and nuclear RNAs to assess effect on MMTV expression.
Results: qRT-PCR on Actinomycin-D-treated MMTV stables are being conducted to determine the effect of mutations in the 5’ and 3’ elements on the expression of genomic as well as spliced RNAs. Mutations in the 5’ element in the presence/absence of RmRE should help clarify the role each element plays in MMTV transcriptional regulation since all studies conducted so far have studied the role of one element in the presence of the other.
Conclusion: MMTV contains a novel cis-acting element downstream of the major splice donor critical for facilitating MMTV gRNA transcription, elongation, transport, and stability. Presence of mirror repeat within the element may represent important viral or host factor binding site(s) within MMTV gRNA.
Acknowledgements: This research was supported by PhD student funds (#31M444) & a grant from the CMHS (#31M331), UAEU.
Authors: Al Mugaddam F (Graduate), Sugathan S, Arnone D, Abdel Aziz K, Karam S. Departments of Psychiatry and Anatomy
Introduction: Ghrelin is a hormone that increases hunger and appetite andhas been implicated in some psychiatric disorders such as dementia and addiction. Ghrelin is involved in mood and reward-related behaviors and Bipolar Disorder (BD) is associated with changes in mood and goal-directed activities, so measuring ghrelin in BD could potentially be significant. To the best of our knowledge, this is the first study in the Arab world to investigate ghrelin levels in BD.
Aims & Objectives: We aimed to measure serum levels of total ghrelin (TG) and active ghrelin (AG) in Bipolar Disorder (BD) subjects compared to healthy controls to determine whether there is an association between ghrelin levels and BD. We hypothesized that there are differences in levels of these markers between subjects with BD and controls.
Materials & Methods: We recruited 24 subjects (15 males, 9 females) diagnosed with BD from the psychiatry department at Al-Ain Hospital, United Arab Emirates between 2016 and 2019. We compared them to 27 healthy controls (13 males, 14 females) for levels of TG and AG. We correlated TG and AG levels with body mass index (BMI) and hunger severity scores using the Hunger Visual Analog Scale.
Results: Mean TG levels in BD were 408.59 pg/mL (SD= 165.49) vs 636.86 pg/mL in controls (SD=295.16) (t= -3.291, p= 0.002) and mean AG levels were 185.22 pg/mL (SD=168.84) in BD vs 521.37pg/mL in controls (SD= 394.17) (t= -3.650, p=<0.0001). AG levels significantly correlated with BMI (r= -0.326; p= 0.024) but not with hunger severity scores (r= 0.152; p= 0.304). TG levels did not significantly correlate with BMI (r= -0.200; p= 0.164) or with hunger severity scores (r= 0.019; p= 0.897).
Conclusion: Our study found significantly lower levels of both TG and AG in BD compared to controls. AG significantly correlated negatively with BMI. This suggests that ghrelin could be a possible marker in BD that is associated with changes in BMI, which itself could be related to medications for the treatment of BD that frequently affect appetite and weight.
Acknowledgments: This study was funded from a seed grant from the CMHS, UAEU.
Authors: Hassan Z. (Graduate), Philip P. S. & Khan G.Department of Microbiology and Immunology, United Arab Emirates University, Al-Ain, UAE.
Introduction: Epstein-Barr virus (EBV) is a ubiquitous herpesvirus associated with human malignancies. It infects and establishes latency in more than 90% of the human population worldwide. Based on the viral genes produced, EBV latency is subdivided into 4 programs, all of which express EBV-Encoded RNAs (EBERs). EBERs are two small capped, non-polyadenylated, non-coding, structurally conserved transcripts that are expressed in millions of copies per infected cell. They have been proposed to have a role in EBV-associated oncogenesis since they promote the rate of cell growth. The mechanisms by which EBERs drive proliferative advantage is poorly understood. Understanding the functions associated with EBERs conserved structure could help immensely in tackling the menace of EBV.
Aims & Objectives: To investigate the structure-to-proliferative function of EBER1 in lymphoid cell line.
Materials & Methods: We disrupted EBER1 structure by deleting individually three of its stem-loops (SLs) to create mutants; DSL1, 3, and 4. These mutants were cloned separately into a Hebo plasmid and expressed in Jurkat cell lines. Wildtype EBER1 and Hebo plasmid transfected cells were used as positive and negative control cells, respectively. A total of 5x104 cells per 2 mL were seeded in a 12-well plate, and cell growth was monitored by trypan blue exclusion dye daily for up to 6 days. Cell cycle progression and regulation were studied using PCR and western blot analyses.
Results: We found a significantly higher cell count and upregulation of the G2/M-phase genes in EBER1 compared to the Hebo, DSL1 and DSL3 but not DSL4 mutants. There was a significant upregulation of G0/G1 and S-phase markers in EBER1 compared to the Hebo cells. We also found differential expression of CDC6, CDC45 and CDT1, among the constructs. Autodegradation of CDT1 protein was observed in Hebo and DSL1 only. Likewise, the absolute cell count of DSL1 was similar to that of Hebo.
Conclusions: EBERs are constitutively expressed molecules in all stages of EBV life cycle. EBER1 conserved structure is crucial for its proliferative advantage derived by prolonging S-phase and disrupting cell cycle regulations.
Acknowledgements: This study wasfunded by grants from Al Jalila foundation (21M132) and Zayed centre-based (31R090).
Authors: Ahmed SAH (Graduate), Anil Kumar C, Kaimala S, Emerald BS. Department of Anatomy.
Background: Studies have shown a link between gestational diabetes mellitus (GDM) and increased risk of developing metabolic syndrome, especially diabetes mellitus (DM). As these changes occur over a long period of time, epigenetic modifications such as DNA methylation are increasingly implicated in these processes.
Aims & Objectives: Our goal is to gain more insight into the epigenetic mechanism (DNA methylation) by which GDM alters fetus’s pancreatic development and predispose the fetus towards metabolic diseases as an adult.
Methodology: Our study investigates the expression of the pancreatic islet hormones, insulin and glucagon, and some of the key transcription factors governing the differentiation and development of the pancreas, aristaless related homeobox (Arx), paired box-6 (Pax6), NK2 homeobox-2 (Nkx2.2), NK6 homeobox-1 (Nkx6.1), and pancreatic and duodenal homeobox-1 (Pdx1), in normal and GDM Wistar rats. Two developmental time points, embryonic days 18.5 (E18.5), and 1-day-old pups where changes in the expression of islet hormones, insulin and glucagon were seen were chosen for analysis.
Results: Immunostaining has revealed significant differences in the protein expression of insulin and glucagon hormones, and the transcription factors, between the two groups, control and GDM, in both of the time points analyzed.
Conclusion: The offspring’s of GDM mothers were heavier at birth and impaired glucose tolerance. They also show changes in the expression of key transcription factors which govern the development and differentiation of the pancreas. Further analysis by using the whole-genome DNA methylation and RNA expression analyses may help in identifying the pathways that might be involved in the increased predisposition of offspring of GDM towards metabolic diseases including GM later in adult life.
Acknowledgements: This work is supported by a Zayd Centre based grant #31R089.
Authors: Hammad, F.T. a, Al-Salam, S b, AlZaabi, S.S.c (Undergraduate), Alfalasi, M. M.c (Undergraduate), Hammad, A.F.d, Yasin, Je, Lubbad, L.a Department of Surgery, b Department of Pathology and e Department of Internal Medicine, College of Medicine & Health Sciences, United Arab Emirates University, Al Ain, United Arab Emirates, c College of Medicine & Health Sciences, United Arab Emirates University, UAE, d School of Medicine, University of Jordan, Amman, Jordan.
Introduction: Renal ischemia-reperfusion injury (IRI) is an invariable consequence of conditions such as renal transplantation. The inhibition of several systems involved in the pathophysiology of the IRI-induced renal alterations have been shown to have beneficial effect on renal functions. This study focuses on the renin-angiotensin system (RAS) and the natriuretic peptide (NP) systems.
Aims & Objectives: Neprilysin (NEP) is the key enzyme for degradation of NPs. The effect of NEP inhibition on the IRI-induced renal alterations has not been investigated and will be investigated in this research. The second aim was to investigate if combining the NEP inhibition to RAS inhibition would have further protective value in IRI.
Materials & Methods: Male Wistar rats underwent bilateral renal ischemia for 20 minutes (G-IRI) and the functional parameters were determined before and after recovery from the IRI. G-Als, G-Scb and G-Als+Scb underwent similar protocol but received Aliskirin (renin inhibitor), Sacubitril (NEP inhibitor) and combination of both pre- and post-IRI, respectively. G-Sham underwent sham surgery and did not receive treatment. Gene expression of renal injury, inflammatory, fibrosis, and apoptotic markers was analysed by real time PCR of kidney tissues.
Results: Before IRI, all renal parameters and markers of renal injury were similar in all groups. Post-IRI, serum creatinine in G-IRI was higher compared to G-Sham. It decreased significantly in G-Als, G-Scb and G-Als+Scb, when compared to G-IRI but not to pre-IRI. Similar trends were observed in serum urea, creatinine clearance and urine albumin/creatinine ratio. Markers of acute renal injury such as kidney injury molecule-1 (KIM-1) was significantly lower in G-Als, G-Scb and G-Als+Scb when compared to G-IRI. Similar findings were obtained for neutrophil gelatinase-associated lipocalin (NGAL) and pro-inflammatory, pro-fibrotic and pro-apoptotic markers such as tumour necrosis factor-alpha (TNF-α), tumour growth factor-β (TGF-β), plasminogen activator inhibitor-1 (PAI-1) and p53. Based on the histology, G-Als+Scb reduced the degree of acute tubular necrosis more significantly than G-Als and G-Scb separately.
Conclusion: RAS blockade by the use of aliskiren or sacubitril separately led to significant attenuation in the renal IRI-induced renal dysfunction. The combination of aliskiren and sacubitril in IRI was more effective than either one alone.
Authors: Parween S (Graduate), Varghesse DS, Ardah MT, Ansari SA. Department of Biochemistry.
Introduction: The nutrient responsive O-GlcNAcylation is a dynamic post-translational proteinmodification found on several nucleocytoplasmic proteins. Previous studies havesuggested that hyperglycemia induces the levels of total O-GlcNAcylation inside thecells. Hyperglycemia mediated increase in protein O-GlcNAcylation has been shownto be responsible for various pathologies including insulin resistance and Alzheimer’sdisease. Since maternal hyperglycemia during pregnancy is associated with adverseneurodevelopmental outcomes in the offspring, it is intriguing to identify the effect ofincreased protein O-GlcNAcylation on embryonic neurogenesis.
Aims & Objectives: To investigate and verify the effect of increased protein O-GlcNAcylation on human embryonic neurogenesis by directed differentiation of hESCs into cortical neurons fate and to investigate how increased O-GlcNAc affect and involve in premature neurogenesis through transcriptional/epigenetic mechanisms.
Materials & Methods: By utilizing the culture system established in our lab hESCs were differentiated from undifferentiated cells (Day0) into neural progenitors (Day12) and early born deep layer cortical neurons (Day63 and Day77) using SMAD inhibition protocol. This process precisely mimics early events of human embryonic corticogenesis which were confirmed by using stage specific markers at different stage of cortical differentiation. Furthermore, during differentiation, cells were treated with vehicle control (DMSO) and 40µM of Thaimet-G (O-GlcNAcase inhibitor) which the level of total O-GlcNac inside the cells and was determined by Immunocytochemistry and western blotting. Further, the expression of various regulatory genes needed for proper embryonic neurogenesis were analysed through western blotting, qPCR and immunocytochemistry and their transcriptional/ epigenetic mechanisms through high-throughput sequencing and CHIP-qPCR.
Results: We discovered that the elevated O-GlcNac modification results in reduced proliferation of neural stem cells and their differentiation into neurons. We also found increased RNA expression of different transcription factors or genes, which are either involved in neuronal differentiation or express in the post-mitotic neurons which in turn might have a role in premature neurogenesis observed.
Discussion: Most neurodevelopmental disorders persist for life and severely impact both the individuals’ and family’s life. In an effort to understand the role of O-GlcNacylation on neurodevelopmental disorders at molecular levels, we propose that increased protein O-GlcNacylation during embryonic neurogenesis could be a mechanisms leading to adverse neurodevelopmental outcomes.
Authors: Bdair R (Graduate), Sulaiman M, Kaimala S, Adeghate E, Mohsin S, Department of Anatomy.
Introduction: Lipocalin 2 (LCN2), also known as neutrophil gelatinase-associated lipocalin (NGAL) plays an important role in the innate immune response and functions as a growth factor. The role of LCN22 as an anti-inflammatory protein and its function in bone metabolism is poorly understood. It is hypothesized that administration of Lipocalin-2 in diabetic and non-diabetic rats will positively improve the bone microstructure.
Aims & Objectives: To investigate the effect of lipocalin2 on bone metabolism in type 1 diabetic osteopathy.
Materials & Methods: Three-month-old male Wistar rats (n=24) were obtained from the animal house facility at United Arab Emirates University for this study. All animal procedures were approved by the animal ethical committee at the United Arab Emirates University. The animals were equally divided into four groups ; (a) control-treated (b) control-untreated (c) diabetic treated (d) diabetic-untreated Animals were made diabetic by injecting them with streptozotocin (STZ) 60 mg/kg and the treated groups were injected with lipocalin-2 (2 µg/ml) for 2 weeks. Bones were dissected out after the animals were sacrificed. The specimens were fixed with 4% paraformaldehyde prior to decalcification using 10% ethylenediaminetetraacetic acid (EDTA) followed by paraffin infiltration and embedding. Histological changes in the proximal part of the femur were analysed using histological stains.
Results: The results of the study showed that type 1 diabetes deteriorates the bone microstructure and bones become porous. LCN-2 treatments improved the bone quality by increasing the thickness of the trabeculae and decreasing the trabecular separation. Our data also exhibited an increase in the size of the growth plate in the treated groups however, changes found were not significant. The treatment improved the number of healthy bone cells and decreased the osteoclasts and mast cells.
Conclusion: Our provisional data results suggest a positive role of LCN2 in bone metabolism, which may be a promising treatment for type 1 diabetic osteopathy.
Acknowledgments: The research was supported by the CMHS faculty grant.
Authors: Hassan Z. (Graduate), Philip P. S. & Khan G. Department of Microbiology and Immunology, United Arab Emirates University, Al-Ain, UAE.
Introduction: Efficient communication among cells is a key factor that coordinates functions and propagates biological processes. One mechanism by which cells perform this vital function is through the release of exosomes. Exosomes are bioactive extracellular vesicles derived from the cell’s endosomal pathway. They are characterised by a lipid membrane and size of 30–120 nm. They regulate physiological and pathophysiological processes. Exosomes contain bioactive molecules, including RNAs, proteins as well as infectious agents and their components. Epstein-Barr virus (EBV) encoded RNAs (EBERs) are two small, capped, non-polyadenylated, untranslated transcripts of RNA polymerase III, expressed at >106 copies per EBV cell. EBERs have been shown to be secreted in exosomes released from EBV infected cells. EBV is an oncogenic herpesvirus associated with many human malignancies of epithelial and lymphoid origins. EBERs are highly conserved in all stages of EBV infection ranging from asymptomatic (latency 0) to malignant (latency III, lytic). It is hypothesised that EBER carrying exosomes induce biological changes that modulate the cellular microenvironment.
Aims & Objectives: To investigate the molecular mechanisms involved in intracellular transport and extracellular secretion of EBER1.
Materials & Methods: We created stable transfectants of EBER1 (wildtype) and EBER1 mutants (EBER1DSL1, DSL3 and DSL4) with deletions in stem-loops shown to interact with cellular proteins that may be involved in intracellular/extracellular movements. Intracellular transport/secretion of the EBER1 was studied using RT-PCR, and qPCR from total cellular, subcellular and extracellular fractions. Western blotting was used to determine the presence of EBER1-binding proteins in the various fractions.
Results: There was a significant reduction in both intracellular and extracellular expression of EBER1 in all mutants compared to the wildtype. However, the levels of EBER1-binding proteins remained similar. Our data suggest that stem-loop 3 of EBER1 maybe the most important loop structure in the secretion of the RNA since deletion of this stem-loop had the greatest impact on the secretion of cytoplasmic EBER1 into exosomes.
Conclusions: Understanding the mechanisms involved in the transport of EBER1 into exosome is important in unravelling the role and function of this abundantly expressed RNA in the biology of EBV and its associated diseases.
Acknowledgements: This study wasfunded by grants from Al Jalila foundation (21M132) and Zayed centre-based (31R090).
Authors: Mehra N1, (Graduate), Challagandla AK, Kaimala S1, Ansari SA2, Al Marzooqi S3, Emerald BS1. Department of Anatomy1, Department of Biochemistry2, Department of pathology3, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, Abu Dhabi, UAE.
Introduction: Pancreatic cancer (PC) is one of the most lethal cancers as it is asymptomatic in the early stages and the fifth most cause of cancer-related deaths including UAE. Pancreatic ductal adenocarcinoma (PDAC) is the most common type, accounting for more than 90% of all PCs. Although the molecular basis of PC development being elucidated the risk factors for PC is not yet fully understood. Hepatocyte nuclear factor 1-beta (HNF1β) encodes a member of the homeodomain-containing superfamily of transcription factors and binds to DNA as either a homodimer or heterodimer with the related protein hepatocyte nuclear factor 1-alpha (HNF1α). The expression of HNF1β is altered in some types of cancer.
Aims & Objectives: Although alteration in the expression of HNF1β has been implicated in some type of cancers its role in PC or PDAC, if any, is not yet known. So we wanted to understand the contribution of HNF1β in PDAC. Such an understanding may help in premalignant diagnoses and may also lead to the development of better intervention strategies for PDAC.
Material & methods: Publicly available RNA/ChIP-Seq data sets were utilized to identify the HNF1β genomic binding locations, respective gene expression patterns and how these are changed in PDAC. From these, we have extracted the metastasis and EMT related pathways to understand the role of HNF1β in PDACs. We have also used PDAC tissue array to verify the expression of HNF1β. Stable ectopic expression of HNF1β and CRISPR/cas9 mediated knockout cell lines are being generated to determine the role of HNF1β in phenotypic conversion (epithelial-mesenchymal transition -EMT) using PDAC cell lines.
Results: The preliminary results from the Oncomine database, PDAC tissue array and PDAC cell lines showed the HNF1β expression was significantly reduced, suggesting a possible role for HNF1β as a tumor suppressor gene in PDAC development. Publicly available RNA-Seq data identified 6410 differentially expressed genes (DEGs) among PANC1, CAPAN2 and CFPAC. We have found the cell migration, cell motility and cell adhesion pathways were enriched based on the DEGs analysis. Based on the ChIP-Seq analysis we have also identified the genes of HNF1β targets and are changed in PDACs.
Conclusion: The preliminary results suggest HNF1β may be a tumor suppressor gene for PDAC.
Acknowledgement: This work is supported by an ADEC grant.