Polysome profiling shows extensive posttranscriptional regulation during human adipocyte stem cell differentiation into adipocytes

Submitted by Manoel Barata ([email protected]) on 2017-08-18T12:45:14Z No. of bitstreams: 1 Spangenbergpoly.pdf: 1003715 bytes, checksum: 4e042c381ca6191b3ccac780cc2def23 (MD5)Approved for entry into archive by Manoel Barata ([email protected]) on 2017-08-18T14:28:45Z (GMT) No. of bitstreams: 1 Spangenbergpoly.pdf: 1003715 bytes, checksum: 4e042c381ca6191b3ccac780cc2def23 (MD5)Made available in DSpace on 2017-08-18T14:28:45Z (GMT). No. of bitstreams: 1 Spangenbergpoly.pdf: 1003715 bytes, checksum: 4e042c381ca6191b3ccac780cc2def23 (MD5) Previous issue date: 2013Institut Pasteur Montevideo. Unidad de Bioinformática. Montevideo, Uruguay.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Núcleo de Tecnologia Celular. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Núcleo de Tecnologia Celular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Institut Pasteur Montevideo. Unidad de Bioinformática. Montevideo, Uruguay.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Adipocyte stem cells (hASCs) can proliferate and self-renew and, due to their multipotent nature, they can differentiate into several tissue-specific lineages, making them ideal candidates for use in cell therapy. Most attempts to determine the mRNA profile of self-renewing or differentiating stem cells have made use of total RNA for gene expression analysis. Several lines of evidence suggest that self-renewal and differentiation are also dependent on the control of protein synthesis by posttranscriptional mechanisms. We used adipogenic differentiation as a model, to investigate the extent to which posttranscriptional regulation controlled gene expression in hASCs. We focused on the initial steps of differentiation and isolated both the total mRNA fraction and the subpopulation of mRNAs associated with translating ribosomes. We observed that adipogenesis is committed in the first days of induction and three days appears as the minimum time of induction necessary for efficient differentiation. RNA-seq analysis showed that a significant percentage of regulated mRNAs were posttranscriptionally controlled. Part of this regulation involves massive changes in transcript untranslated regions (UTR) length, with differential extension/reduction of the 3'UTR after induction. A slight correlation can be observed between the expression levels of differentially expressed genes and the 3'UTR length. When we considered association to polysomes, this correlation values increased. Changes in the half lives were related to the extension of the 3'UTR, with longer UTRs mainly stabilizing the transcripts. Thus, changes in the length of these extensions may be associated with changes in the ability to associate with polysomes or in half-life

Polysome profiling shows the identity of human adipose-derived stromal/stem cells in detail and clearly distinguishes them from dermal fibroblasts

O artigo encontra-se disponível em acesso aberto no site do Editor.Submitted by Michel Batista ([email protected]) on 2014-12-11T23:53:17Z No. of bitstreams: 1 zych2014 (1).pdf: 669092 bytes, checksum: c4215d46bb3118c318f1160caf288d86 (MD5)Approved for entry into archive by Michel Batista ([email protected]) on 2014-12-12T00:08:23Z (GMT) No. of bitstreams: 1 zych2014 (1).pdf: 669092 bytes, checksum: c4215d46bb3118c318f1160caf288d86 (MD5)Made available in DSpace on 2014-12-12T00:08:23Z (GMT). No. of bitstreams: 1 zych2014 (1).pdf: 669092 bytes, checksum: c4215d46bb3118c318f1160caf288d86 (MD5) Previous issue date: 2014This work was supported by grants from Ministério da Saúde and Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq, FIOCRUZ-Pasteur Research Program, and Fundação Araucária. L.S. received fellowship from ANII (Agencia Nacional de Investigación e Innovación, Uruguay); S.G., J.Z., and B.D. from CNPq; P.S. from FIOCRUZ; and A.C. from Fundação Araucária.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Instituto Pasteur. Unidad de Bioinformática. Montevideo, Uruguay.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Núcleo de Tecnologia Celular. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Núcleo de Tecnologia Celular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Instituto Pasteur. Unidad de Bioinformática. Montevideo, Uruguay.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Although fibroblasts and multipotent stromal/stem cells, including adipose-derived stromal cells (ADSCs), have been extensively studied, they cannot be clearly distinguished from each other. We, therefore, investigated the cellular and molecular characteristics of ADSCs and fibroblasts. ADSCs and fibroblasts share several morphological similarities and surface markers, but were clearly found to be different types of cells. Contrary to previous reports, fibroblasts were not able to differentiate into adipocytes, osteoblasts, or chondrocytes. Polysome-bound mRNA profiling revealed that*1,547 genes were differentially expressed (DE) in the two cell types; the genes were related to cell adhesion, the extracellular matrix, differentiation, and proliferation. These findings were confirmed by functional analyses showing that ADSCs had a greater adhesion capacity than fibroblasts; the proliferation rate of fibroblasts was also higher than that of ADSCs. Importantly, 185 DE genes were integral to the plasma membrane and, thus, candidate markers for ADSC isolation and manipulation. We also observed that an established marker of fibroblasts and ADSCs, CD105, was overexpressed in ADSCs at both mRNA and protein levels. CD105 expression seemed to be related to differentiation capacity, at least for adipogenesis. This study shows that ADSCs and fibroblasts are distinct cell types. These findings should be taken into account when using these two cell types in basic and therapeutic studies