The role of microRNAs on the proliferation and cell differentiation of the embryonal carcinoma pluripotent cell line NTera-2.

• Grant number: 13/27061-0
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: May 01, 2014
• End date: September 30, 2017
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Rodrigo Alexandre Panepucci
• Grantee: Ildercílio Mota de Souza Lima
• Host Institution: Hemocentro de Ribeirão Preto. Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da USP (HCMRP). Secretaria da Saúde (São Paulo - Estado). Ribeirão Preto , SP, Brazil
• Associated research grant: 13/08135-2 - CTC - Center for Cell-Based Therapy, AP.CEPID
• Associated scholarship(s): 15/15864-6 - Functional evaluation of microRNAs in cell proliferation and differentiation of embryonal carcinoma cell line NTera-2 using high content screening, BE.EP.DR

Abstract

The pluripotency is the capacity that some cells posses to ability to generate cells from the three germ layers (ectoderm, mesoderm and endoderm) as well as the germ cells. The embryonic stem cells (ESC) are the most known pluripotent cells, they present a high capacity of cell differentiation and self-renewal. These properties make ESC as potential tools for regenerative medicine, but its use in pratical clinic is not viable, because, in addition to the ethics issues, it is necessary a imunocompatibilite between these cells and the host. Aiming to solve these problems, it was generated the induced pluipotent stem cells (iPSC). These cells, on other hand, are obtained thought nuclear reprogramming by inserting some pluripotency-related factor, initially been these factor the ones known as the four Yamanaka Factors (Oct4, Sox2, c-Myc and Klf4). It was recently discoverd some regulatory RNAs, the microRNAs (miRNAs) that plays an important role on pluripotency maintaining, as well as on cell differentiation process. These miRNAs are small molecules of RNA that act as pós-transcriptional regulators of gene expression by binding to target mRNA resulting in translation inhibition. In addition to pluripotency and cell differentiation process, these miRNAs may regulate many cellular processes like proliferation and cell death among others. It is known that the use of synthetic miRNAs molecules can generate iPSC or increase the efficiency in nuclear reprogramming, as well as to induce stem cells differentiation. The embryonal carcinoma cells (ECC) have similarities to ESC, like pluripotency, in vitro cell differentiation capacity and potential for generate teratomas in vivo. Comparative analysis between the profile of mRNA and miRNA expression in ESC and ECC demonstrated these cells share similarities, thus the use of ECC is widely applied in studying the pluripotent stem cells biology. The present study aims to evaluate the effect of different miRNAs on a human ECC line, NTera-2, by using synthetic precursors (pré-miRs) or inhibitors (anti-miRs) miRNAs molecules, studying the effects of these molecules over cell proliferation, apoptosis and cell differentiation through using different methodologies as qPCR, microarrays and High-Content Analysis/Microscopy (HCS).

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)

PANEPUCCI, RODRIGO ALEXANDRE; DE SOUZA LIMA, ILDERCILIO MOTA. Arrayed functional genetic screenings in pluripotency reprogramming and differentiation. STEM CELL RESEARCH & THERAPY, v. 10, JAN 11 2019. (15/15864-6, 13/08135-2, 15/08070-3, 13/27061-0)

DE SOUZA LIMA, ILDERCILIO MOTA; DOS SANTOS SCHIAVINATO, JOSIANE LILIAN; PAULINO LEITE, SARAH BLIMA; SASTRE, DANUTA; DE OLIVEIRA BEZERRA, HUDSON LENORMANDO; SANGIORGI, BRUNO; CORVELONI, AMANDA CRISTINA; THOME, CAROLINA HASSIBE; FACA, VITOR MARCEL; COVAS, DIMAS TADEU; et al. High-content screen in human pluripotent cells identifies miRNA-regulated pathways controlling pluripotency and differentiation. STEM CELL RESEARCH & THERAPY, v. 10, JUL 8 2019. (15/08070-3, 15/15864-6, 13/27061-0, 13/08135-2)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

LIMA, Ildercílio Mota de Souza. Identification of microRNA-modulated pathways in cell differentiation and pluripotency maintainance in human cells. 2017. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC) Ribeirão Preto.