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Identification of single-nucleotide polymorphisms in microRNAs involved with the glioblastoma susceptibility

Grant number: 15/03870-1
Support Opportunities:Regular Research Grants
Duration: November 01, 2015 - October 31, 2018
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Manoela Marques Ortega
Grantee:Manoela Marques Ortega
Host Institution: Universidade São Francisco (USF). Campus Bragança Paulista. Bragança Paulista , SP, Brazil
Associated researchers: Ander Matheu Fernandez ; Carmen Silvia Passos Lima


Glioblastoma multiforme (GBM) is the most common primary malignant brain tumor. Patients with GBM have a median survival of about 14 months, despite aggressive treatment. Due to the clinical and pathological heterogeneity of the GBM, there are many recent attempts to better understand and characterize these tumors at the genetic and molecular level. The goal of this study is to identify single nucleotide polymorphisms (SNPs) in microRNAs (miRNAs) as risk factors for GBM. To achieve this goal, it will be performed, for the first time, a global analysis of about 278,000 SNPs located in regions of miRNAs in 80 DNA samples collected from tumor tissues of newly diagnosed GBM patients and in 80 DNA samples collected from non-tumor brain tissues of healthy subjects undergoing surgery to remove brain tumor by using Axiom® miRNA Target Site Genotyping Array (Affymetrix). Next, a selection of relevant SNPs in GBM compared with samples from healthy individuals will be performed. After the selection of candidate SNPs in miRNAs as risk factors for the disease, the influence of different genotypes (homozygous wild allele, heterozygous allele and variant allele) will be evaluated by quantitative real-time PCR (qPCR) using RNA samples extracted from other 200 RNA samples collected from tumor tissues of newly diagnosed GBM patients. Also, qPCR will be performed using RNA samples collected from at least 10 fresh primary cultures derived from fresh tumors tissue obtained from newly diagnosed GBM patients after surgery and 60 frozen tumors tissue obtained from newly diagnosed GBM patients after surgery. RNA samples collected from normal brain tissue of patients undergoing surgery to remove brain tumor will be used as control. After studying the influence of different genotypes in the precursor of miRNAs selected above, a relevant miRNAs whose function has not been defined previously will be selected for functional studies. The binding of the selected miRNAs to untranslated region (UTR) of the mRNA of the predicted target gene generated using TargetScan, will be tested. Also, it will be possible evaluate the impact of the SNPs on miRNA expression of the target gene expression. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
BONAFE, GABRIEL ALVES; DOS SANTOS, JESSICA SILVA; ZIEGLER, JUSSARA VAZ; UMEZAWA, KAZUO; RIBEIRO, MARCELO LIMA; ROCHA, THALITA; ORTEGA, MANOELA MARQUES. Growth Inhibitory Effects of Dipotassium Glycyrrhizinate in Glioblastoma Cell Lines by Targeting MicroRNAs Through the NF-kappa B Signaling Pathway. FRONTIERS IN CELLULAR NEUROSCIENCE, v. 13, . (15/03870-1, 17/03064-0)

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