InCited2 secrete cardiogenic molecules

• Autor(a) principal: Lopes, João André Pereira
• Data de Publicação: 2020
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10400.1/15097
• topic: CITED2; FGF10; Secretoma; Efeitos cardíacos; Células estaminais embrionárias; Zebrafish; Domínio/Área Científica::Ciências Médicas::Outras Ciências Médicas

The CBP/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 2 (Cited2) interacts with many CBP/p300-dependent transcriptional factors. Cited2 plays an important role in cardiac development in humans and mouse. Furthermore, in vitro studies using embryonic stem cells (ESC) have demonstrated its importance for their specification into cardiac cell lineages. Moreover, the WNT5a and WNT11 were identified and overexpressed in the conditioned medium of ESC overexpressing CITED2 (CM-CITED2) and showed to hold the potential to rescue cardiogenic defects caused by Cited2 depletion in vitro and in zebrafish embryos. We believe that more secreted proteins which may be influenced by CITED2 expression may also have the potential to rescue the cardiogenic defects caused by Cited2 depletion. Amongst these potential candidate proteins FGF10 was identified in a preliminary study but was not validated for its capacity to reverse cardiac defects in zebrafish triggered by Cited2 depletion. Here, we have assessed the capacity of FGF10 to rescue defects caused by Cited2 depletion in zebrafish. Although, we were not able to definitively conclude about FGF10’s ability to rescue defects caused by Cited2 depletion due to a possible degradation of the morpholinos (MO) used to inhibit Cited2 expression, we obtained some promising preliminary results. In order to identify other candidate proteins that may be present in the secretome of CITED2 overexpressing cells, we have used Mass Spectrometry analysis to further determine the content of CM-CITED2 and control medium (CM-control). Interestingly, we identified a differential composition of proteins in the conditioned media, and amongst these proteins, some have been described to be associated with cardiomyocyte’s development. The analysis of transcripts expression of genes encoding relevant proteins suggested that these proteins may be expressed at critical cardiogenic steps. Overall, this work demonstrates that overexpression of CITED2 may promote the early onset expression of genes normally expressed in cardiac differentiated cells. And though we were not able to conclude about FGF10 ability to rescue Cited2 deficiency, we identified other secreted proteins that may as well hold a potential for the development of therapies