Department Of Human Genetics And Molecular Medicine

Permanent URI for this communityhttps://kr.cup.edu.in/handle/32116/103

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End date: 2020Has files: No

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    Pharmacogenetics of Cyclophosphamide and Doxorubicin in Breast Cancer & Synthesis and Biological Evaluation of New Imidazole Based Putative Anticancer Agents
    (Central University of Punjab, 2019) Kalra, Sourav; Munshi, Anjana & Kumar, Raj
    Breast cancer is the second most prevalent cancer in women worldwide. It is treated by various strategies, including surgery, radiation therapy, hormonal therapy, targeted therapy and chemotherapy. Chemotherapeutic drugs like cyclophosphamide, doxorubicin, paclitaxel and docetaxal are used to treat the cancer either alone or in combination with other therapies, such as surgery, radiation, or hormone therapy depending upon the stage and the location of cancer cells. Cyclophosphamide and doxorubicin are also given in combination known as adjuvant chemotherapy (AC therapy). However, all the patients do not get benefited by this treatment. Inter-individual response in patients on AC therapy might be on account of variation in genes involved in pharmacokinetics and pharmacodynamics of cyclophosphamide and doxorubicin. Therefore, a detailed pharmacogenomic study was performed using global screening array microchip that contains more than 700,000 upto-date markers, optimized for human genome-wide backbone for unparalleled genomic coverage, including clinically relevant content and all pharma GKB markers. Two gene variants CYP2C19*2 and ALDH1A1*2 involved in the metabolism of cyclophosphamide were found to influence the clinical outcome in breast cancer patients on AC therapy in Malwa region of Punjab. However, none of the gene variants involved in pharmacokinetics vi and pharmacodynamics of doxorubicin were found to be involved in the interindividual response. Therefore, patients should be screened for CYP2C19*2 and ALDH1A1*2 gene variants before prescribing the AC therapy. Due to several problems with the chemotherapeutic drugs the new therapeutic strategies have been introduced in the recent past that have led to the target based drug design. In an attempt to develop new drugs for targeting breast cancer, imidazole based anti-cancer agents were computationally designed and synthesized (Series 1 and Series 2). All the compounds were first assessed for their anti-proliferative activity in T47D, MDA-MB-231, and MCF7 breast cancer cell lines. Lead compounds with IC50 values less than 10 µM in at least two cell lines such as A12, SA1, SA3, SA8, SA11 from series 1 and SRA1, SRA8, SRA12, SRA14 from series 2 were selected for further evaluation of their cytotoxicity (if any) against normal HBL100 and HPBMC cell lines. Two compounds SRA12 and SRA14 were found to be non toxic and upon lead optimisation through in silico approach at CDK4/6 protein afforded ANP12 that emerged out to be potent anticancer compound and exhibited good anti- breast cancer profile. Compounds ANP12 and SRA14 (from series 2) elevated ROS levels, altered mitochondrial membrane potential and induced cell cycle arrest at G0 phase indicating their anticancer mechanism possibly via CDK4/6 inhibiton.
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    Epidemiological factors, genetic and inflammatory markers in the development of Breast Cancer in Malwa region of Punjab
    (Central University of Punjab, 2019) Kaur, Raman Preet; Munshi, Anjana
    Breast cancer is one of the most common malignancy among women and also the leading cause of death worldwide. Malwa region of Punjab is reported to have the highest number of cancer cases in comparison with the other two regions (Doaba and Majha). The cancer prevalence in this region was reported to be 1089 per million/year in 2013 (with around 35.7% breast cancer cases), as per the Department of Health and Family Welfare. The aim of present study was to carry out a systematic epidemiological study, explore the contribution of mutations in breast cancer susceptibility genes, and gene-gene interactions in the development of breast cancer in Malwa region of Punjab. In addition, exome sequencing of the male breast cancer patients was carried out to identify the specific genetic alterations associated with the disease in this region. Inflammatory markers (cytokines and C-reactive protein levels) and albumin levels were also estimated to evaluate their association with the development of the disease and outcome including death, recurrence, and metastasis. Three hundred breast cancer patients (Female:Male = 296:4) were recruited from Guru Gobind Singh Medical College & Hospital, Faridkot and Max Hospital, Bathinda. Equal number of age and sex matched controls were also recruited from the same demographic area with the help of Rural Medical Officers. The study was approved by the ethics committee of the Central University of Punjab. The information regarding the demographic profile was collected in a structured questionnaire. The blood samples were collected with the written informed consent of the patients. DNA isolation was carried out by phenol:chloroform method. The genotyping was performed by global screening array (GSA; Illumina Infinium HD), exome sequencing and PCRRFLP. The levels of inflammatory markers and albumin were estimated by using BD Accuri flow cytometer and ERBA 200 bioautoanlyzer. The association of demographic features with the disease was evaluated by Student’s t-test. Mann-Whitney test was used to evaluate the association of CRP, cytokines and albumin with the outcome. Softwares including R/Bioconductor and OpenEpi were used to evaluate the association of pathogenic alterations with the disease. The influence of novel mutations on protein structure was examined by molecular dynamics simulations. Follow-up of the patients was carried out with the help of clinicians. Infiltrating breast cancer was the most common type of breast cancer in patients. Triple negative breast cancer was observed in 17.5% of patients. All the patients underwent modified radical mastectomy and lumpectomy. The breast cancer patients v carried mono/biallelic alteration in 15 breast cancer susceptibility genes including ATM, BRCA2, STK11, PTEN, BRIP1, CHEK2, RAD51C, NBN, MLH1, MSH2, MSH6, MUTYH, CDK2NA, CYTB and MTHFR as per the results of GSA. Familial cases of breast cancer were found to carry alterations in ATM, BRCA2, STK11, PTEN, BRIP1, CHEK2, RAD51C, NBN, MLH1, MSH2, MSH6, MUTYH, CDK2NA, CYTB and MTHFR genes. Novel pathogenic variants in PALB2, STK11, CHEK2, BRCA2, BARD1, BRIP1, NF2, and FZR1 genes were detected in male breast cancer patients. These were found to alter the specific protein structure. Among the inflammatory markers CRP, IL-6 & IL17A were found to be significantly associated with poor outcome in the patients. High levels of albumin were also found to be associated with increased overall survival. In conclusion, results of this study can guide to develop a panel to test the breast cancer patients for pathogenic mutations from Malwa region of Punjab. The present study suggests that the screening of BRCA2, MSH2, and MLH1 should be carried out in general population since previous studies have reported that these mutations increase the risk of breast cancer by 10-fold and accordingly evidence-based management recommendations can be given to the individuals bearing these mutations. Familial breast cancer patients need to be screened for BRAD1, MSH2, MLH1, BRCA2 and CYTB genes since these alterations were observed in familial breast cancer patients. IL-6, IL-17A, CRP, and albumin can be used as prognostic markers for breast cancer in Malwa region of Punjab.
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    Transcriptional Regulation of Pro-metastatic Protein Formin Binding Protein17 (FBP17) in Breast Cancer
    (Central University of Punjab, 2018) Suman, Prabhat; Chander, Harish
    Breast cancer is a diverse disease with multiple subtypes. Among the different molecular subtypes, triple negative breast cancers (TNBC) harbor frequent mutation in tumor suppressor p53. Recently it was shown that p53 suppresses Transducer of Cdc42-dependent Actin assembly-1 (Toca-1) that belongs to CIP4 subfamily. Members of the family including FBP17 play a significant role in actin assembly. FBP17 and Toca-1 have been recognized as key scaffolds for recruiting actin regulatory protein to promote invadopodia formation. Metastatic cancer cells form invadopodia and the F-BAR proteins are shown to enhance invadopodia. FBP17 consists of F-BAR domain, Cdc-15 homology, putative Rho-binding domain and SH3 domain. In the present study, we elucidate the correlation between p53 and FBP17 that affects metastatic potential of cancer cells. We observed that cancer cell lines with mutated p53 have high levels of FBP17. Activation of wild type p53 reduces FBP17 both at mRNA and protein level. Further, the ectopic expression of wild type p53 reduces FBP17 whereas mutant p53 failed to do so. Different cell lines and different methods of p53 activation were used to study the p53-FBP17 axis. Chromatin immunoprecipitation studies revealed the binding of p53 in the promoter of FBP17. The metastatic potential of breast cancer cells was observed after double knock down of both p53 and FBP17. Interestingly, we found that combined silencing of these two proteins led to a partial rescue of invasion upon p53 silencing in vitro. In conclusion we suggest that p53 controls FBP17 expression and FBP17 contributes to the invasion of cancer cells upon loss of p53 activity in cancer.
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    Mechanisms of Anti-Tumor Activity of Withania somnifera (Ashwagandha)
    (Routledge, 2020) Mehta, V; Chander, H; Munshi, A.
    Increasing herbal formulations have been used to treat several diseases including cancer. W. somnifera (Ashwagandha) is one such plant the extracts of which have been tested against a number of ailments including cancer, which remains as one of the most dreadful diseases on the globe. The ever-increasing number of cancer related mortality demands the development of novel chemopreventive agents with minimum side effects. Different compounds isolated from various parts of the plant like root, stem, and leaves have been reported to display significant anti-cancerous and immunomodulating properties and thus can be used alone or in combination with other chemotherapeutic drugs for cancer treatment. Through this review, we highlight the importance of W. somnifera in countering the potential oncogenic signaling mediators that are modulated by active constituents of W. somnifera in a variety of cancer types. Further, we hope that active constituents of W. somnifera will be tested in clinical trials so that they can be used as an important adjuvant in the near future for the effective treatment of cancer. � 2020, � 2020 Taylor & Francis Group, LLC.
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    Combinatorial Effect of DCA and Let-7a on Triple-Negative MDA-MB-231 Cells: A Metabolic Approach of Treatment
    (SAGE Publications Inc., 2020) Sharma, P; Singh, S.
    Dichloroacetate (DCA) is a metabolic modulator that inhibits pyruvate dehydrogenase activity and promotes the influx of pyruvate into the tricarboxylic acid cycle for complete oxidation of glucose. DCA stimulates oxidative phosphorylation (OXPHOS) more than glycolysis by altering the morphology of the mitochondria and supports mitochondrial apoptosis. As a consequence, DCA induces apoptosis in cancer cells and inhibits the proliferation of cancer cells. Recently, the role of miRNAs has been reported in regulating gene expression at the transcriptional level and also in reprogramming energy metabolism. In this article, we indicate that DCA treatment leads to the upregulation of let-7a expression, but DCA-induced cancer cell death is independent of let-7a. We observed that the combined effect of DCA and let-7a induces apoptosis, reduces reactive oxygen species generation and autophagy, and stimulates mitochondrial biogenesis. This was later accompanied by stimulation of OXPHOS in combined treatment and was thus involved in metabolic reprogramming of MDA-MB-231 cells. � 2020 Sage Publications.
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    miRNA dysregulation in ischaemic stroke: Focus on diagnosis, prognosis, therapeutic and protective biomarkers
    (Blackwell Publishing Ltd, 2020) Vasudeva, K; Munshi, A.
    Stroke is one of the leading causes of death and disability in both developing and developed countries. Biomarkers for stroke and its outcome can greatly facilitate early detection and management of the disease. miRNAs have been explored for their potential as biomarkers for diagnosis, prognosis and brain injury in ischaemic stroke. A substantial body of evidence suggests that miRNAs play key roles in numerous cellular changes following ischaemic stroke including mitochondrial dysfunction, energy failure, cytokine-mediated cytotoxicity, oxidative stress, activation of glial cells, increased intracellular calcium levels inflammatory responses and disruption of the blood�brain barrier (BBB). In addition, targeting specific miRNAs, therapeutic modulation of brain injury and apoptosis can also be achieved. Therefore, the current review has been compiled within an aim to give an overview of the developments exploiting miRNAs at different stages of stroke as prognostic, diagnostic, protective and therapeutic biomarkers. � 2020 Federation of European Neuroscience Societies and John Wiley & Sons Ltd
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    Establishing molecular signatures of stroke focusing on omic approaches: a narrative review
    (Taylor and Francis Ltd, 2020) Ludhiadch, A; Vasudeva, K; Munshi, A.
    Stroke or �brain attack� is considered to be the major cause of mortality and morbidity worldwide after myocardial infraction. Inspite of the years of research and clinical practice, the pathogenesis of stroke still remains incompletely understood. Omics approaches not only enable the description of a huge number of molecular platforms but also have a potential to recognize new factors associated with various complex disorders including stroke. The most significant development among all other omics technologies over the recent years has been seen by genomics which is a powerful tool for exploring the genetic architecture of stroke. Genomics has decisively established itself in stroke research and by now wealth of data has been generated providing new insights into the physiology and pathophysiology of stroke. However, the efficacy of genomic data is restricted to risk prediction only. Omics approaches not only enable the description of a huge number of molecular platforms but also have a potential to recognize new factors associated with various complex disorders including stroke. The data generated by omics technologies enables clinicians to provide detailed insight into the makeup of stroke in individual patients, which will further help in developing diagnostic procedures to direct therapies. Present review has been compiled with an aim to understand the potential of integrated omics approach to help in characterization of mechanisms leading to stroke, to predict the patient risk of getting stroke by analyzing signature biomarkers and to develop targeted therapeutic strategies. � 2020, � 2020 Informa UK Limited, trading as Taylor & Francis Group.
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    Correction to: Association of the genetic variants of insulin receptor substrate 1 (IRS-1) with type 2 diabetes mellitus in a Saudi population (Endocrine, (2014), 47, 2, (472-477), 10.1007/s12020-014-0177-2)
    (Springer, 2020) Alharbi K.K.; Khan I.A.; Munshi A.; Alharbi F.K.; Al-Sheikh Y.; Alnbaheen M.S.
    Type 2 diabetes mellitus (T2DM) is a chronic degenerative disease, phenotypically and genetically heterogeneous, characterized by high levels of glucose and metabolic complications. Insulin receptor substrate 1 (IRS-1) plays a key role in the insulin-stimulated signal transduction pathway. A glycine-to-arginine substitution at codon 972 (G972R) (rs1801278) in the IRS-1 gene has been associated with impaired insulin action. Another SNP rs2943641 in the IRS-1 gene has been found to be associated with T2DM and insulin resistance in genome-wide association studies. The aim of the present study was to evaluate whether rs1801278 and rs2943641 are associated with increased risk of T2DM in the Saudi population. The study included 376 T2DM cases and 380 healthy controls. Genomic DNA was isolated using a commercially available kit supplied by Norgen Biotech Corp. Genotyping was performed by PCR and RFLP analysis. There was a significant difference in the genotypic distribution as well as allelic frequency between the T2DM cases and controls in case of both the polymorphisms for rs1801278 (1.752, 95 % CI 1.002–3.121; p = 0.04), and for rs2943641 (OR = 1.482, 95 % CI 1.176–1.867; p = 0.001). In conclusion, both the (rs1801278 and rs2943641) polymorphisms are associated with T2DM in the Saudi population.
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    Small regulatory molecules acting big in cancer: Potential role of mito-miRs in cancer
    (Bentham Science Publishers, 2019) Sharma P.; Bharat; Dogra N.; Singh S.
    MicroRNAs [miRNAs] are short, non-coding, single stranded RNA molecules regulating gene expression of their targets at the posttranscriptional level by either degrading mRNA or by inhibiting translation. Previously, miRNAs have been reported to be present inside the mitochondria and these miRNAs have been termed as mito-miRs. Origin of these mito-miRs may either be from mitochondrial genome or import from nucleus. The second class of mito-miRs makes it important to unravel the involvement of miRNAs in crosstalk between nucleus and mitochondria. Since miRNAs are involved in various physiological processes, their deregulation is often associated with disease progression, including cancer. The current review focuses on the involvement of miRNAs in different mitochondrial mediated processes. It also highlights the importance of exploring the interaction of miRNAs with mitochondrial genome, which may lead to the development of small regulatory RNA based therapeutic options.
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    Genetics of platelet traits in ischaemic stroke: focus on mean platelet volume and platelet count
    (Taylor and Francis Ltd, 2019) Vasudeva K.; Munshi A.
    Purpose/Aim of the study: The aim of this review is to summarize the role of genetic variants affecting mean platelet volume (MPV) and platelet count (PLT) leading to higher platelet reactivity and in turn to thrombotic events like stroke and cardiovascular diseases. Materials and Methods: A search was conducted in PUBMED, MEDLINE, EMBASE, PROQUEST, Science Direct, Cochrane Library, and Google Scholar related to the studies focussing on genome-wide association studies (GWAS), whole exome sequencing (WES), whole genome sequencing (WGS), phenome-wide association studies (PheWAS) and multi-omic analysis that have been employed to identify the genetic variants influencing MPV and PLT. Results: Antiplatelet agents underscore the crucial role of platelets in the pathogenesis of stroke. Higher platelet reactivity in terms of mean platelet volume (MPV) and platelet count (PLT) contributes significantly to the interindividual variation in platelet reaction at the site of vessel wall injury. Some individuals encounter thrombotic events as platelets get occluded at the site of vessel wall injury whereas others heal the injury without occluding the circulation. Evidence suggests that MPV and PLT have a strong genetic component. High throughput techniques including genome-wide association studies (GWAS), whole exome sequencing (WES), whole genome sequencing (WGS), phenome-wide association studies (PheWAS) and multi-omic analysis have identified different genetic variants influencing MPV and PLT. Conclusions: Identification of complex genetic cross talks affecting PLT and MPV might help to develop novel treatment strategies in treating neurovascular diseases like stroke. � 2019, � 2018 Informa UK Limited, trading as Taylor & Francis Group.