Banca de DEFESA: MARIANA BORGES LOPES

Uma banca de DEFESA de DOUTORADO foi cadastrada pelo programa.
STUDENT : MARIANA BORGES LOPES
DATE: 30/03/2020
TIME: 09:00
LOCAL: Centro de Estudos da MEJC
TITLE:

Modulation of the expression of RNAms through the use of miRNA mimics: gene therapy applied to Diabetic Cardiomyopathy” no Periódico “Cardiovascular Research

KEY WORDS:

Diabetes mellitus; cell line H9c2; animal model; mimic miRNAs; Pla2g2a; miR-214 Biomarkers; gene theraphy.

PAGES: 148
BIG AREA: Ciências da Saúde
AREA: Farmácia
SUMMARY:

Diabetic cardiomyopathy (DCM) is a chronic complication of diabetes with high rates of mortality and morbidity. In this context, the present study aimed, innittialy, through an in silico analysis, mRNAs and miRNAs involved in the development of DCM, followed by validation in vitro and in vivo of this molecules, and to elucidate the role of Pla2g2a gene and it possible regulatory miRNA rno-miR-214-3p in inflammation caused by hyperglycemia, proposing an alternative to treatment of DCM based in gene therapy by miRNA mimics. In silico analyzes using microarray datasets available on Gene Expression Omnibus (GSE4745 and GSE44179), tools of Ingenuity Pathway Analysis and miRNAs database of TargetScan made possible an integrative analysis between mRNAs, miRNAs and DCM. The in vitro study, cell culture from myoblast lineage H9c2 were plated in normoglycemic (NG, 5.5 mmol/L of glucose) and hyperglycemic (HG, 25 mmol/L of glucose), extraction of total RNA from the cells has been performed to analyze gene expression (miRNAs e mRNAs) by real time PCR (RTqPCR). In the in vivo model, Wistar rats had diabetes pharmacologically induced by streptozotocin (40 mg/kg, i.v.; n=7) and was compared to the control group (citrate buffer, n=7), after 30 days from induction the animals were euthanized and the left ventricle (LV) was used for total RNA and protein extraction for the analysis of mRNAs and miRNAs expression by RTqPCR and proteins by Western Blotting. Pla2g2a gene was upregulated on the online microarray datasets¸ in the H9c2 cell cultura under hyperglycemic conditions (3-fold increased, p-value=0.043) and in LV of diabetic rats (3-fold increased, p-value=0.004). The rno-miR-214 was reduced both in the databases and in the analysis of in vitro (p-value= 0.043) and in vivo (p=0.025) studies. The expression of sPLA2IIA and IL-6 proteins were increased (p = 0,008 for both proteins) in the LV of diabetic rats. Still in vitro, was performed the transfection of rno-miR-214-3p mimic (50nM for 24 hours) increasing its expression by 6000 times, RTqPCR Pla2g2a and Il6 gene expression results showed that this genes tends to decrease (p-value > 0,005). These results suggests that use bioinformatic tools could assist in the search of biomarkers of diseases. As well, demonstrates that the use of rno-miR-214-3p mimics can be considered an innovative technique for controlling the translation of Pla2g2a mRNA into sPLA2-IIA, modulating Il6 expression, reducing DCM effects.

BANKING MEMBERS:
Externo à Instituição - ALVARO DANILO CERDA MAUREIRA - UFRO
Externa à Instituição - ALICE CRISTINA RODRIGUES - USP
Presidente - 2087759 - ANDRE DUCATI LUCHESSI
Interno - 2275890 - MARCELO DE SOUSA DA SILVA
Interno - 1544647 - MATHEUS DE FREITAS FERNANDES PEDROSA