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Characterization of embryonic stem cell transcriptome during its differentiation into insulin-producing cells in the absence and presence of growth/differentiation peptide factors

Grant number: 19/21935-4
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2020
Effective date (End): January 31, 2024
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Mari Cleide Sogayar
Grantee:Isaura Beatriz Borges Silva
Home Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Associated research grant:16/05311-2 - Regenerative medicine aiming at therapy for chronic degenerative diseases (cancer and diabetes), AP.TEM
Associated scholarship(s):21/12497-3 - Evaluation of a pectin-polymer and human recombinant prolactin in improving the viability and functionality of microencapsulated pancreatic islets, BE.EP.DR

Abstract

Diabetes mellitus (DM) is a chronic disease characterized by the absence of insulin production (T1D) or inefficiency of its use by the individual (T2D). Consequently, patients with DM develop hyperglycemia, which, if not controlled, can lead to serious complications in the functioning of vital organs. Conventional T1D treatments are often unable to maintain perfect control of glycemic levels. Given the growing need for new therapeutic interventions for T1D, strategies based on stem cell differentiation into insulin producing cells have been shown to be a promising approach. The present work proposes to analyze the transcriptomic profile during the process of differentiation of murine Embryonic Stem Cells (mESCs) and genetically modified mESCs, in which the Txnip gene is inhibited, into Insulin Producing Cells (IPCs), in the presence of different growth and differentiation peptide factors. Using standardized methodologies by our laboratory, the cell differentiation process will be performed in the absence and presence of peptide factors (PDGF-BB and BMP-7), aiming to identify the differentially expressed gene sets using large-scale RNAseq transcriptome analysis. Subsequently, the most relevant transcripts during the differentiation process will be selected to perform molecular interventions (overexpression and/or knockout) in a functional genomics approach. This approach should allow to reveal the molecular basis of the mESCs differentiation process into IPCs and to identify new molecular targets acting in this process. The results obtained will be critically important for the development of better strategies for DM treatment, aiming to minimize the risk of complications and even death and to improve the quality of life of patients with unstable T1D. (AU)

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