Day 1 :
Keynote Forum
Manohar R Furtado
Apton Biosystems Inc., USA
Keynote: Super resolution imaging system for DNA sequencing, genotyping and proteomics
Time : 09:00 - 10:00
Biography:
Manohar R Furtado has completed his PhD in Protein Biochemistry from Pune Universsity and completed Post-Doctoral Training at Northwestern University in Virology. He served on the Faculty at Northwestern in the Department of Pathology and conducted Basic Research in Oncolgy & Infectious Diseases. He joined Applied Biosystems in 2000 and worked there as the Vice Presisident, R&D for Molecular Medicine & Applied Markets. He was a Consultant for Bio-Rad, ACD, Sample 6, DxNow and Apton Biosystems. He was on the National Biodefense Science Board at DHHS (2011-2015). He is currently the CSO and CRO at Apton. He is on the Board of LexaGene.
Abstract:
Apton Biosystems has built a versatile high-speed, affordable, imaging system with the ability to detect single fluorophores on labeled macromolecules (DNA, RNA and protein) on a surface with super resolution capability. The system was used for DNA sequencing using commercial sequencing by synthesis reagents dNTPs and Therminator X enzyme and for molecule detection using fluorescently labeled oligonucleotide and antibogy probes. Target molecules or clusters with incorporated labeled dNTPs or bound by probes or have are imaged and countedWe have sequenced PCR amplified human gene targets, Phage X174 genome, the E. coli genome human exomes and the human genome. We will present a cost validated execution plan to reduce the sequencing costs to $ 10 per human genome.. NSCLC cell lines were cultured and untreated or treated with tyrosine kinase inhibitor erlotinib. We detected protein phosphorylation changes for EGFR, ERK, MET and MEK, using just 0.5 to 2 cell equivalents of protein lysate containing sub pM levels of protein. We also detected indels in EGFR exon 19 and point mutations in L858R, T790M in EGFR and V600E in BRAF, at 0.5% minor component levels, by using an oligonucleotide ligation assay off-chip and then attaching the ligated product to the surface. mRNA levels and fusion mRNAs were detected using 10-20 cell equivalents of RNA. The system will enable comprehensive analysis of cancer related pathways from a few cells to help decipher changes in cellular pathways in response to mutations and consequently help with selection of efficacious drugs that are individual specific. We believe this system will revolutionize the practice of medicine by enabling affordable analysis of biological samples, cellular mutations and pathways.
Keynote Forum
Pravin D Potdar
Dr. A P J Abdul Kalam Education & Research Centre, India
Keynote: Personalized medicine: Rays of hopes to cure of cancer
Time : 10:00 - 11:00
Biography:
Pravin D Potdar has completed his PhD from Tata Memorial Centre, Mumbai in 1991. He has been working in the field of Cellular and Molecular Biology of Cancer. He has retired from Jaslok Hospital & Research Centre, Mumbai as a Head, Department of Molecular Medicine& Biology, after servicing there for 12years. He has established Molecular Diagnostics & Stem cell Research Laboratories at Jaslok and carried out various research Program in the field of Cancer Genetics and developed innovated technologies which can help to diagnose and treat various cancers. He has published 81 papers in national and international journals. He is an Editorial Board of various journals. He is a recipient of National Cancer Institute, USA award and also holds Faculty position at M D Anderson Cancer Centre; Houston TX, USA, He is currently working at Dr A.P.J. Abdul Kalam Research Institute, India.
Abstract:
Cancer is among the leading cause of death all over the world. Every year millions of cancer patients are enrolled for cancer treatment in which only few patients survive with present cancer therapy. Since last two decades, the revolution has occurred in health care industries. Completion of human genome project by Dr. Francis Collins has introduced the molecular profiling of individual cancer patient to diagnose cancer more precisely than any other method. It has also allowed clinicians to prescribe appropriate therapies which match with cancer patient’s own profile. This new form of medicine is termed as a personalized medicine. Worldwide, personalized medicine programs have focused on analysis of genetic profile of cancer patients by using whole genome sequencing of cancer patients to get precise advice for cancer treatment. However, it is understood that the outcome of cancer treatment is not determined only by the variation in the genetic makeup of a tumor but also inter patient variations in pharmacokinetics which may give rise to drug resistant. Therefore, personalized medicine requires not only to characterize tumor cells but also to see the individualized drug resistant effect on these patients. Protein kinase activity is significantly increased in most of cancers and thus the protein kinase inhibitors play very important role in treatment of various cancers. 17 small molecules and 4 antibodies are approved as protein kinase inhibitors, as cancer therapeutic drugs. These drugs interact with specific site of cancer cells and bring out significant inhibition of cancer cell growth. Most of these drugs are focused on only 8 common linkage targets i.e. VEGF or VEGFR, PI3K, EGFR2 (HER2), mTOR, EGFR, MET, PDGF or PDGFR and KIT. Presently HER2 antibody Trastuzumab is used in HER2 positive breast cancer patients. Similarly, BCR-ABL1 inhibitor Imatinib is used in chronic myeloid leukaemia. Whereas, Gefitinib and Erlotinib which are EGFR kinase inhibitors are used in Non-Small-Cell Lung Cancer (NSCLC). BRAF inhibitor Vemurafenib is used in melanoma and the dual ALK-MET inhibitor Crizotinib is used in NSCLC. Kinase inhibitors for cancer therapy are very much dependent on predictive biomarkers expressed on cancer patient’s cells. Thus the presence or absence of these predictive markers is directly linked to the response rates of particular targeted therapies with small-molecule kinase inhibitors or antibodies. Similarly, latest innovation in analysis of liquid biopsy is also given a non-invasive platform for doing molecular profiling of cancer patients without taking tissue biopsy. Next-generation sequencing potentiates the sequencing of circulating tumor cells or cell-free DNA in liquid biopsies of cancer patients which helps in diagnosis and therapy of cancer. So over all, we are confident that in near future we can be able to implement personalized medicine treatment to counteract with the cellular and molecular heterogeneity of cancer and its drug resistance to cure the cancer.
- Human Genetics | Gene Mutation | Genetic Diseases| Cancer Genetics | DNA Sequencing| Medical Genetics | Stem Cell Therapy | Bioinformatics and Computational Biology | Genome Editing | Sickle Cell Anemia | Genomics | Regenerative Medicine
Location: Andromeda | Radisson Blu Hotel, Abu Dhabi Yas Island
Chair
Mohammed Chyad Al-Noaemi
Al-Ghad International College, Saudi Arabia
Session Introduction
Asma M Alshammari
Kuwait Medical Genetic Centre, Kuwait
Title: Massively parallel next generation sequencing to investigate the cis-acting genetic modifiers of somatic instability in Huntington’s disease
Biography:
Asma M Alshammari is currently working as a senior specilisit in Human Molecular Genetics at Kuwait Medical genetics Centre. She has completed her PhD in Human Molecular Genetics from University of Glasgow.
Abstract:
Huntington Disease (HD) is an extremely variable inherited neurodegenerative disorder caused by expansion of an unstable CAG trinucleotide repeat in the huntingtin gene (HTT). Somatic instability in HD exhibits an age-dependent, expansion-biased and tissue-specific pattern and the highest level of somatic instability is found in tissues that are most susceptible to the disease pathology. Therefore, the aim of this project was to test the hypothesis that somatic instability of the HD CAG repeat plays a major role in disease pathology by quantifying somatic instability in the number of CAG repeats by next generation sequencing (NGS) technology in buccal cell DNA. We developed a high-throughput sequencing pipeline to sequence and genotype HTT alleles from blood and buccal swab DNA of the Scottish and Venezuelan populations, respectively. A total of 210 individuals from the Scottish general population and 742 HD patients and unaffected individuals from the Venezuelan HD cohort were sequenced on the MiSeq platform. We established that it was possible to sequence and genotype the CAG repeats, the polymorphic CCG repeat and the flanking sequences. Our data highlight the utility of NGS technology as an approach to genotype HTT alleles, detect sequence variants and quantify somatic instability of the CAG repeat. Our data emphasise that the somatic instability in HD is age-dependent and expansion-biased, also could be a major factor in disease progression and could be a potential therapeutic target in HD.
Mariam Fida et al.
Al Jawhara Center, Bahrain
Title: Allele amplification failure in the HBB gene due to allelic dropout in a pre-preimplantation genetic testing case
Biography:
Mariam Fida has recieved her PhD in Medical Genetics from the Univesrity of Edinburgh, Scotland. She is also specialized in Preimplantation Genetics Diagnosis at Reproductive Genetic Innovations in Chicago. She established the first PGD center in Bahrain and is currently the Director of the Center and is a Consultant in Medical Genetics.
Abstract:
Beta-Thalassemia is one of the common autosomal recessive inherited genetic conditions in the GCC region due to increased rates of consanguinity. It is a condition caused by several different types of mutations in the HBB gene. The current case represents a 30-year old gentleman who was informed to be a heterozygous carrier of β-thalassemia. At the clinic, peripheral blood was drawn for DNA sequencing. Although the patient indicated that he was a carrier, sequencing results surprisingly showed him to be affected with the codon 39 mutation which does not correspond at the genotype-phenotype level indicating the possibility of an allele dropout. Further analysis was carried out using two different sets of designed primers to confirm the carrier status. This showed that the gentleman is indeed a carrier of β-thalassemia at codon 39 of the HBB gene. In conclusion, to overcome and minimize the incidence of an allele dropout event in the pre-PGT setting and during PGT-M, it is advisable to detect every mutation using at least two different sets of primers to avoid unknown SNPs around the mutation region.
Hilde Van Esch
Katholieke Universiteit Leuven, Belgium
Title: Modelling neuro developmental disorders in a dish
Biography:
Hilde Van Esch has completed her PhD from Katholieke Universiteit Leuven, Belgium. She has then pursued Post-doc at Institut Cochin in Paris on X-linked intellectual disability. She currently works as a Clinical Geneticist at the University Hospitals UZ Leuven and is Associate Professor in the Department of Human Genetics and Head of the Laboratory of Genetics of Cognition at the University of Leuven, Belgium. Her research interests includes the identification of genes involved in rare diseases and intellectual disability. She has published and coauthered more than 170 papers in reputed journals and is a Board Member of the European Society of Human Genetics.
Abstract:
The leading manifestation of brain dysfunction is intellectual disability, affecting approximately 3% of the general population. Given the uniqueness and the complexity of human cognition and behavior, studies in humans are essential to understand the role of the multitude of genes involved in these processes. We previously developed IPSC from patients with MECP2 duplication syndrome carrying different duplication sizes, to study the impact of increased MePC2 dosage in human neurons. MECP2 duplication syndrome is a severe neurodevelopmental disorder in males, characterized by severe neurodevelopmental delay with onset at birth, limited or absent speech, hypotonia, epilepsy, autism and motor dysfunction. Cortical neurons derived from Mecp 2dup-iPSCs had more synapses and altered network synchronization as well as dendritic complexity. Next, we tested a series of epigenetic drugs for the ability to rescue neuronal defects and validated two HDAC inhibitors as potential clinical candidates. We are currently developing an iPSC model for a novel tubulinopathy, characterized by intellectual disability associated with characteristic dysmorphic signs: circumferential skin creases, cleft palate, facial dysmorphisms and short stature. This developmental disorder is caused by mutations in a novel gene, MAPRE2 (Microtubule-Associated Protein Member 2) encoding a member of the EB family. We derived patient-specific induced Pluripotent Stem Cell (iPSC) and established isogenic rescue lines as well as patient specific knock-in lines using CRISPR/CAS9. These iPSC’s are then differentiated towards Neural Progenitor Cells (NPC), cortical neurons and Cranial Neural Crest Cells (CNCC) using adapted and optimized differentiation protocols. Preliminary functional experiments using patient fibroblasts showed an increase in migration speed and overall mitosis duration. We will now perform similar experiments to analyze the mitosis and migration rate of iPSC derived NPC’s and CNCC’s. In addition a full morphological work-up is ongoing. We will present these novel data at the meeting.
Manisha Mishra et al.
All India Institute Of Medical Sciences, India
Title: Association of MMP2 polymorphisms in preeclampsia
Biography:
Manisha Mishra had completed Bachelor of Dental Surgery and MSc Anatomy from All India Institute of Medical Sciences, New Delhi, India. She is currently a Senior Reseach Fellow at All India Institute of Medical Sciences, India.
Abstract:
Background & Aim: Imbalanced Matrix Metalloproteinase (MMP) expression, including MMP-2, has been demonstrated in pre-eclampsia. However, little is known about the effect of MMP-2 gene polymorphisms on hypertensive disorders of pregnancy. Therefore, we examined Matrix Metalloproteinase (MMP-2) gene polymorphisms (g.-735C/T) and its association with Preeclampsia (PE) and measured the levels of MMP-2 serum concentrations in PE. Method: 30 preeclamptic and 30 healthy pregnant women were enrolled from department of obstetrics and gynaecology, AIIMS, New Delhi after getting approval from Institute ethical committee. Genomic DNA was extracted from blood and amplified by PCR. MMP2 gene polymorphisms of -735C/T was detected by Restriction Fragment Length Polymorphism (RFLP). The levels of MMP2 in sera were measured by ELISA. Result: The maternal serum MMP2 levels was found to be more in PE patients than in control group (p=0.03). The increased frequency of CT genotype for MMP2 (-735C/T) Single Nucleotide Polymorphism was seen in PE patients as compared to control group. However the difference in genotype frequency was not statistically significant (p=0.35).
Conclusion: These findings may help to understand the relevance of MMP-2 and its genetic polymorphisms to the pathophysiology of hypertensive disorders of pregnancy like preeclampsia and IUGR.
Sarah Sabir
Kinnaird College for Women, Pakistan
Title: Molecular characterization of families affected with autosomal recessive primary microcephaly
Biography:
Sarah Sabir has completed her MPhil (Master in Philosophy) in Biochemistry in 2016 from Kinnaird College, Pakistan. Her research interests are in molecular genetics of various diseases, retrovirus biochemistry, neurodevelopmental disorders, cell division, cell biology and structural biological studies of proteins. She has published her Bachelors research work entitled “Molecular Studies on preproinsulin gene” in Matec web of conference 2016 6th International Conference on Chemistry and Chemical Process, ICCCP 2016. She also did Internship at Chagatai’s Lahore lab for six months and worked in the departments of hematology, microbiology, molecular biology and biochemistry and learned about testing of blood samples, streaking and culturing of the samples of blood and urine, doing PCR on HCV samples and performing LFTs and RFTs on various blood samples. Currently she is working as a High School Science Assistant in Lahore American School. She has plans to pursue her PhD in Biochemistry from a well known institute.
Abstract:
Statement of the Problem: Cousin Marriage is very common in part of world and there have been proven association with the degree of consanguineous marriages and prevalence of autosomal recessive genetic disorders. It is reported that 60% of cousin marriages in Pakistan is the subject of prevalence of most rare genetic disorders in our population. Microcephaly which is present at birth and causes non-progressive mental retardation is called autosomal recessive primary microcephaly (MCPH), whereas which develops postnatally is secondary microcephaly. It is a neurogenic mitotic disorder which results in small brain size compared to one third of normal brain. Affected patients have normal neuronal migration, neuronal apoptosis and neuronal function. Sixteen MCPH loci have been reported by different scientist from various populations around the world containing the following genes Microcephalin, WDR62, CDKRAP2, CASC, ASPM, CENPJ, STIL, CEPH135, CEPH152, ZNF335, PHC1, CDK6, CENPE, SAS6, MFSD2A and ANKLE2. Mutations in any one of the gene will lead to disease phenotype due to premature chromosomal condensation, disturbed mitotic spindle orientation, signaling response as a result of DNA damage.
Methodology & Theoretical Orientation: In the current study, the molecular genetics of five families affected with autosomal recessive primary microcephaly from Pakistani origin were studied. By taking the head circumference of the affected individual these families were identified from different cities of Pakistan. Clinical information was collected with the help of a carefully designed questionnaire. Venous blood sample of the affected families was collected and genomic DNA was extracted using standard phenol-chloroform method. Four loci i.e. MCPH5 (ASPM), MCPH2 (WDR62), MCPH1 (Microcephalin) and MCPH6 (CENPJ) were selected for initial screening bring the most prevalent loci reported from Pakistan. Linkage analysis of the affected families was done by Polymerase Chain Reaction (PCR) using specific microsatellite markers flanking the selected gene. 8% native Polyacrylamide Gel (PAGE) was used to identify PCR results.
Conclusion & Significance: Five families affected with autosomal recessive primary microcephaly were mentioned in this study. These families were screened for ASPM, WDR62, CENPJ genes. These families showed no linkage to the initial screening done for the three most prevalent loci reported from this region. Hence, it can be concluded that disease phenotype in these families is apparently not due to mutations in these three genes, however further screening using more markers and mutation screening of these families will give confirmed results. The aim of this study is to elucidate the molecular genetics of this disorder in five affected families. Linkage analysis will be initially done followed by mutations screening of the families linking to any of the known loci. The current project will enable us to offer carrier screening and genetic counseling, which will be our meager contribution towards reducing the prevalence of this disease our parent population. Results: The gel analysis revealed that two of the families are linked with ASPM gene, whereas the rest of three were not found linked with ASPM, WDR62, CENPJ and Microcephalin genes.
Asmaa AlFadil Hassan Khalifa
University of Medical Sciences and Technology, Sudan
Title: Genetic markers associated with inflammatory bowel disease - detection of NODI, 2 mutations in inflammatory bowel disease among Sudanese patients
Biography:
Biography Asmaa AlFadil Hassan Khalifa has completed her BSc from University of Gezira School of Pharmacy and MSc studies from University of Medical Sciences and Technology, Sudan. In 2018, she has completed a Diploma in IV Compounding Techniques from Notting Hill College, Manchester, United Kingdom. She is a Clinical Pharmacy Specialist and a Member of Sudan Medical Council. She is licensed as Clinical Pharmacist in Department of Health, Abu Dhabi, United Arab Emirates also a BLS Provider in American Heart Association. She did a research in Injection Safe Disposal: Safe Disposal Policy in Compliance with WHO recommendations in 2010 for her BSc thesis sponsored by WHO. Late in 2013 she presented her second research: Genetics Markers associated with Inflammatory Bowel Disease: Detection of NOD1,2 Mutations in Inflammatory Bowel Disease among Sudanese Patients, in completion of her thesis for MSc degree. Through her professional journey she worked as Community Pharmacist, House-Officer Pharmacist, Hospital In-Charge Pharmacist and Clinical Pharmacist. She joined many facilities while she was in Sudan which include: Paediatric Hospital, Oncology Institute, Khartoum Teaching Hospital and Police Hospital. In UAE she worked in Tawam Hospital as a Trainee and lately worked for Canadian Medical Centre.
Abstract:
Inflammatory Bowel Disease (IBD) comprising Ulcerative Colitis (UC) and Crohn's Disease (CD) is debilitating chronic immune disorder of the intestinal mucosa, multigenic in nature, resulting from dysfunctional interactions between the intestinal immune system and its micro flora, influenced by host genetic susceptibility as suggested in linkage, epidemiologic, racial, familial aggregation and twin studies. It has led to the discovery of mutations in nucleotide- binding oligomerization domain containing protein 2 (NOD2) also known as caspase recruitment domain containing protein 15 (CARD15) or inflammatory bowel disease protein 1 (IBD1) which located in chromosome 16q12 and associated with ileal CD, also numerous other genes’ mutations have been found to be associated with IBD susceptibility such as NOD1/CARD4 mutations in chromosome 7p14.3 in UC patients. Nod-like receptor family plays a key role in realization of innate and adaptive immune response. Their polymorphisms may shift balance between pro- and antiinflammatory cytokines, modulating the risk of chronic inflammation. The rational and prevalence of IBD is dependent on geographic location and racial background. The study met its objective in determining whether genetic, environment and socio-economical components contribute to the development of IBD in Sudanese population and furthermore, whether the race and dietary intake associated. Different groups’ joined this study were from Sudanese population. They had been divided into two; patients with colonoscopy diagnosed IBD and individuals who don’t have IBD as controls. Each group was classified according to gender, age and race. Blood samples (Biospecimen Retention) from the included participants at the Gastroenterology Department in Ibn-Sina Hospital were collected and sent to Al-Neelain Medical Research Centre for DNA purification and isolation, Polymerase Chain Reaction (PCR) amplification, establishment of sequence analysis and product identification of genetic polymorphisms associated with IBD. Finally genotyping was the end point. Also, a questionnaire was designed to collect data in such a way to achieve objectives of the study. The collected data was tabulated and analyzed using Statistical Package for Social Sciences (SPSS 14.0) computer program. The study revealed that Shaigeia and Ga’aleia (Northern tribes) were found to be more affected by Ulcerative Colitis than other tribes (13.3%). Thus, suggesting possible link with ethnic origin. Also, it resulted that ulcerative colitis was associated with the economic status of the patient. Subjects with low economic status were found to be more likely to develop it (60%). The most important result after the laboratory experiments was the obvious association of ulcerative colitis with the allelic mutation in NOD1 (70%). While Crohn’s disease was not found to be associated with (-/C) allele insertion.
Nigora Mavlyanova et al.
Ministry of Health of the Republic of Uzbekistan, Uzbekistan
Title: The role of xenobiotic enzyme genes in predicting fetal loss syndrome in Uzbekistan
Biography:
Nigora Mavlyanova is the Scientific Leader of the Youth Applied Grant from the Republican Specialized Scientific and Practical Medical Center of Obstetrics and Gynecology, Ministry of Health of the Republic of Uzbekistan. She is a Member of the Association of Obstetricians and Gynecologists of the Republic of Uzbekistan. She has published over 20 articles in famous magazines.
Abstract:
Introduction: Fetal loss syndrome is a multifactorial disease characterized by a universal integrated response of the female body to any ill health in the pregnant woman, the fetus and the environment, the result of the action of functionally weakened variants (alleles) of many genes against the background of adverse external and internal factors. Glutathione-S-transferase (GST) metabolizes foreign substances or controls the entry of carcinogens into cells. Aim: The goal of our research was to establish the role of the polymorphic variants of the xenobiotic enzymes genes GSTM1 and GSTT1 and IIe 105Val of the gene GSTP1 in the mechanism of formation and development of fetal loss syndrome. Material & Methods: Molecular genetic studies were conducted in 114 pregnant women aged from 20 to 45 years old. Molecular genetic examination of biomaterials (DNA) was performed at Department of Molecular Medicine and Cellular Technologies of the Research Institute of Hematology and Blood Transfusion of the Ministry of Health of the Republic of Uzbekistan. Statistical analysis of the results was carried out using the statistical software package "OpenEpi 2009, Version 2.3".
Results: The results of molecular genetic studies in pregnant women with Fetal Loss Syndrome (PPS) showed an increased detection rate of combined functionally defective genotypes GSTM10/0+GSTT10/0 - 25.4%, against the control group 4.1% (χ2=12.4; P=0.0004; OR=7.8; 95% CI 2.146-28.65). Whereas, with the combined variants - the null and functional genotypes of the polymorphism of the GSTM1 and GSTT1 genes between the studied groups did not reveal statistically significant differences (χ2=0.1; P=0.3; OR=1.4; 95% CI 0.697-282; p>0.05). Whereas, the distribution of genotypes IIe 105 Val of the GSTP1 xenobiotic enzyme in pregnant women revealed a high detectability of A/G genotype polymorphism in the pregnant group compared to the control group 56.1% versus 19.4%, which was 2.9 times higher than the control groups. Thus, an analysis of the association of genic combinations of zero polymorphisms of the GSTM1 and GSTT1 genes revealed that in the group of pregnant women with fetal loss syndrome, combinations of the homozygous del/del genotype responsible for the lower level of protein product synthesis are significantly more common. The chance of developing pathology in the presence of this combination of the genotypic version of the del/ del genes GSTM1 and GSTT1 increases significantly: Up to 7.8 times more than other genotypes (χ2=12.4; P=0.0004; OR=7.8; 95% CI 2.146-28.65). Whereas, the functionally unfavorable GSTP1 G allele 2.7 times was statistically significantly predominant in the studied chromosomes of pregnant women with PPS compared with pregnant women without PBS (χ2=4.6; P=0.03; OR=4.5; 95% CI 1.061-19.5).
Conclusion: Analysis of the results showed that the polymorphism variants of the GSTM10/0+GSTT10/0 genotypes of the GSTM1 and GSTT1 genes, as well as the G/A IIe 105 Val genotypes of the GSTP1 gene are significant predictors of the risk of developing fetal loss syndrome, resulting in disorders of the detoxification process in the body in women during pregnancy.
Salma Ahmadloo
Shahid Beheshti University, Iran
Title: Gene expression signatures identify novel regulatory genes and signaling pathways in young hypertensive patients making them susceptible to develop premature coronary heart disease
Biography:
Salma Ahmadloo completed her PhD the field of Human Genetics at University Putra Malaysia, currently she is postdoctoral fellowship at Shahid Beheshti University, Iran. She is an experienced senior researcher with a demonstrated history of working in higher education industry, skilled in gene expression profiling, PCR array technique, real time PCR and Genotyping.
Abstract:
Hypertension stands out as major modifiable risk factors for premature Coronary Heart Disease (CHD) and it is well established that the incidence of over 80% of CHD is attributable to the modifiable risk factors including hypertension. This chronological study was carried out for the purpose of profiling expression of hypertension associated genes and identify related biological process and modulated signaling pathways of young Malaysian subjects (<55 years old, total number= 168) with hypertension who had developed premature CHD after at least five years. In order to achieve the goal, four groups of subjects were divided into: (1) 42 Healthy subjects, (2) 42 subjects with only HT, (3) 42 subjects with only HT and (4) 42 subjects with CHD+HT. The RNA was extracted from blood specimens by mean of commercial extraction kits. The RT2 Profiler™ PCR Array was utilized to determine gene profiling on group 1 and group 2, group 1 and group 3, group 1 and group 4. To validate the results of RT2 Profiler™ PCR Array, significantly dysregulated genes were selected and validation was conducted through q-RT-PCR in a larger and independent population. For this purpose, new subjects (total number=150) were divided into: (1) 75 Healthy subjects, 2) 75 subjects with hypertension+CHD. Sixteen significantly dysregulated genes related to hypertension were identified and the Ras cell signaling pathway was highlighted as a culprit in people suffering from hypertension which may be prone to CHD. In silico analysis showed that the majority of the identified genes involved in reninangiotensin regulation and other categories related to renin-angiotensin such as, regulation of blood volume and regulation of blood vessel by renin-angiotensin. In conclusion, some differentially dysregulated genes and modulated pathways were identified which warrant further investigation in the setting of premature hypertension and CHD.