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Large-scale analysis of microRNA function in cell cycle, pluripotency, self-renewal and differentiation of stem cells using high-content screening

Grant number: 15/08070-3
Support type:Scholarships abroad - Research
Effective date (Start): December 01, 2015
Effective date (End): June 30, 2016
Field of knowledge:Health Sciences - Medicine
Principal Investigator:Rodrigo Alexandre Panepucci
Grantee:Rodrigo Alexandre Panepucci
Host: Marc Bernard Thomas Bickle
Home 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
Local de pesquisa : Max Planck Society, Dresden, Germany  
Associated research grant:13/08135-2 - CTC - Center for Cell-Based Therapy, AP.CEPID

Abstract

Pluripotency and self-renewal are distinctive properties of embryonic stem cells (ESCs). microRNAs (miRs) play important roles in the post-transcriptional control of gene expression, by targeting multiple mRNA, and have been implicated in the maintenance of pluripotent cells characteristics. Many miRs effects are attributed to cell cycle regulation, in line with the intimate link between cell-cycle regulation, self-renewal, pluripotency and differentiation. We will systematically investigate the roles of all currently annotated human miRs (2,588 mature miRs) in these characteristics, using automated High-Content Screening fluorescence microscopy. Images derived from pluripotent cells transfected with microRNA mimics will be analyzed, in order to quantify biological features reflecting, cell cycle status (based on Cyclin B1 staining and Hoechst/DNA content analysis), pluripotency (based on Oct4 staining) and differentiation (cell morphology). Single-cell data will be combined into multiparametric population profiles describing miR induced phenotypes. Profiles will be clustered to identify microRNA groups with common effects. Enrichment functional analysis of predicted miR targets will be carried out to identify biological pathways and processes mediating the phenotypes observed. The primary objective of the present FAPESP-BPE application is to allow the applicant to develop specific image and data analysis pipelines, under the guidance of Dr. Marc Bickle (Max Planck Institute of Molecular Cell Biology and Genetics, Dresden-Germany). For this, images from a pilot focused screen (30 miRs) will be used in the development and optimization of the analysis pipelines. Once established, the pipelines will be adapted to the full library screen, which will be carried by the PhD student Ildercílio Lima (supervised by Dr. Panepucci, FAPESP 2013/27061-0), in collaboration with Dr. Miguel Mano (ICGEB, Trieste-Italy). In view of the complexity of the analytical approaches, this fellowship will bring important immediate and precisely defined benefits for the success of this project (a subproject of the Center for Cell Therapy - CTC, a CEPID FAPESP).

Matéria(s) publicada(s) na Agência FAPESP sobre a bolsa:
New mechanisms that regulate pluripotency in embryonic stem cells are discovered 

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)
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; ZAGO, MARCO ANTONIO; GIACCA, MAURO; MANO, MIGUEL; PANEPUCCI, RODRIGO ALEXANDRE. High-content screen in human pluripotent cells identifies miRNA-regulated pathways controlling pluripotency and differentiation. STEM CELL RESEARCH & THERAPY, v. 10, JUL 8 2019. Web of Science Citations: 0.
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. Web of Science Citations: 1.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.
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