Functional evaluation of microRNAs in cell proliferation and differentiation of embryonal carcinoma cell line NTera-2 using high content screening

• Grant number: 15/15864-6
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: October 15, 2015
• End date: April 14, 2016
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Rodrigo Alexandre Panepucci
• Grantee: Ildercílio Mota de Souza Lima
• Supervisor: Miguel Luis Cunha Mano
• 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
• Institution abroad: International Centre for Genetic Engineering and Biotechnology (ICGEB), Italy
• Associated to the scholarship: 13/27061-0 - The role of microRNAs on the proliferation and cell differentiation of the embryonal carcinoma pluripotent cell line NTera-2., BP.DR

Abstract

Pluripotency and the ability to self-renew are distinctive properties of embryonic stem cells (ESCs), that confer them a great potential in regenerative medicine. The understanding of the regulatory networks acting on ESCs, led to the identification of a core set of transcription factors (TFs) able to control pluripotency, leading to the discovery that somatic cells could be reprogramed into induced pluripotent stem cells (iPSCs). In addition, small RNA molecules called microRNAs (miRNAs) have been shown to effect on gene expression by binding to target mRNAs, leading to translation repression and mRNA degradation and some of them are specifically expressed in pluripotent stem cells (PSC). The ectopic expression of these miRNAs can improve the generation of iPSCs. Although a lot of work has been devoted to the characterization of miRNAs differentially expressed in undifferentiated cells and their differentiated counterparts, there is a lack of systematic studies aiming at defining the roles of the known miRNAs in the biology of PSC. Only a handful of works evaluated the function of a few hundred miRNAs, on the proliferation of ESCs or in the reprograming process. Despite the reduced number, the aforementioned studies helped to pinpoint important miRNA functions in the regulation of self-renewal, pluripotency and differentiation of mouse PSC. Functional studies are fundamental to elucidate the roles of miRNAs in the biological properties of PSC and to accelerate the progress in this important research field. With that in mind, we will systematically investigate the roles of all the currently annotated human miRNAs in the regulation of pluripotency and cell proliferation in an embryonal carcinoma pluripotent cell line (NTera-2), using unbiased experimental approaches based on High Content Screening by fluorescence microscopy. Overall this project will lead to an unprecedented understanding of the role of miRNAs in pluripotency and cell proliferation by a functional screening of human miRNAs. (AU)

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)