Crosstalk between Hedgehog pathway and energy pathways in human adipose-derived stem cells: A deep sequencing analysis of polysome-associated RNA

Submitted by Manoel Barata ([email protected]) on 2019-02-14T14:01:07Z No. of bitstreams: 1 s41598-018-265.pdf: 2868417 bytes, checksum: 2aafcd778e3da201b5a4ff508d60ebbc (MD5)Approved for entry into archive by Manoel Barata ([email protected]) on 2019-02-21T15:11:16Z (GMT) No. of bitstreams: 1 s41598-018-265.pdf: 2868417 bytes, checksum: 2aafcd778e3da201b5a4ff508d60ebbc (MD5)Made available in DSpace on 2019-02-21T15:11:16Z (GMT). No. of bitstreams: 1 s41598-018-265.pdf: 2868417 bytes, checksum: 2aafcd778e3da201b5a4ff508d60ebbc (MD5) Previous issue date: 2018Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células Tronco. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células Tronco. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Grupo de Espectrometria de Massas Computacional e Proteômica. Curitiba, PR, Brasil.Pontifícia Universidade Católica do Paraná. Escola de Medicina. Programa de Graduação em Ciências da Saúde. Divisão de Terapia Celular. Laboratório Experimental Multiuso. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células Tronco. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Básica de Células Tronco. Curitiba, PR, Brasil.Adult stem cells are considered promising candidates for cellular therapies due to their capacity to differentiate and self-renew. Differentiation leads to changes in the metabolism, structure, and gene expression patterns of cells. Hedgehog is one of the pathways that is involved in the enhancement of osteogenesis and chondrogenesis in adult stem cells, but its mechanisms are poorly understood. In this study, we treated adipose tissue-derived stem cells (ADSC) with two well-characterized drugs, purmorphamine (Hedgehog pathway activator) and cyclopamine (Hedgehog pathway inhibitor), and identified mRNAs associated with polysomes in each treatment group to determine the post transcriptional genetic networks governed by the Hedgehog pathway. Activation of the Hedgehog pathway by purmorphamine results in significant upregulation of mRNAs associated with cellular communication and signal transduction. Furthermore, our experiments show that cyclopamine acts late downregulating GLI1 expression in ADSCs but promotes the upregulation of mRNAs associated with energy pathways and metabolism at early times. Through in silico analysis, we identified some miRNAs, such as miR-355, that could regulate these mRNAs association with polysomes and thereby modulate the Hedgehog pathway. Our results suggest that activation of the Hedgehog pathway by purmorphamine also results in a negative regulation of mRNAs in the protein translation machinery