Splicing regulators in endothelial cell differentiation

Concetta Schiano, Monica Rienzo, Amelia Casamassimi, Andrea Soricelli, Claudio Napoli

Research output: Contribution to journalArticle

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Abstract

Aims Alternative splicing represents a key mechanism of gene regulation. Despite its role in regulating cell pluripotency and differentiation being well known, the underlining mechanisms are still poorly studied. Here, we investigated the possible involvement of splicing regulators during the different steps of endothelial cell differentiation through expression studies on human circulating progenitors. Methods Total RNAs were extracted from all cells and reverse-transcribed. Semiquantitative and real-time RT-PCR was performed using selective oligonucleotides. Differences between group means were considered significant at P value less than 0.05 and more significant at P value less than 0.01. Protein extracts were incubated with an antibody directed against MED23. Immunoprecipitation of supernatants and pellets was probed with both anti-Muscleblind-like splicing regulator (MBNL)1 and anti-MBNL2 antibodies. Results Several clinical trials demonstrated the safety and efficacy of progenitor cells in regenerative therapy of the cardiovascular system. Particularly, we analyzed the expression of genes belonging to muscleblind family members and MED complex subunits, which are known to be involved during differentiation in other models. This study shows that MED23, MBNL1 and MBNL2 were all expressed at high levels only in differentiated cells. Moreover, immunoprecipitation assays indicated that MED23 is able to bind MBNLs in endothelial cells. Conclusion Our data suggest that MED23, MBNL1 and MBNL2 could regulate alternative splicing events activated during differentiation through a common mechanism. Hence, these observations corroborate previous evidence that splicing regulators may have an essential role in the basic apparatus required for cell pluripotency and reprogramming, allowing identification of novel biomarkers to use for early diagnosis in cardiovascular diseases.

Original languageEnglish
Pages (from-to)742-749
Number of pages8
JournalJournal of Cardiovascular Medicine
Volume18
Issue number10
DOIs
Publication statusPublished - Oct 1 2017

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Alternative Splicing
Immunoprecipitation
Cell Differentiation
Endothelial Cells
Cardiovascular System
Oligonucleotides
Real-Time Polymerase Chain Reaction
Early Diagnosis
Anti-Idiotypic Antibodies
Cardiovascular Diseases
Stem Cells
Biomarkers
Clinical Trials
RNA
Safety
Gene Expression
Antibodies
Genes
Proteins
Therapeutics

Keywords

  • alternative splicing
  • cell therapy
  • human muscleblind-like family
  • mediator complex

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Splicing regulators in endothelial cell differentiation. / Schiano, Concetta; Rienzo, Monica; Casamassimi, Amelia; Soricelli, Andrea; Napoli, Claudio.

In: Journal of Cardiovascular Medicine, Vol. 18, No. 10, 01.10.2017, p. 742-749.

Research output: Contribution to journalArticle

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