Cripto Is Targeted by miR-1a-3p in a Mouse Model of Heart Development
During cardiac differentiation, numerous factors contribute to the development of the heart. Understanding the molecular mechanisms underlying cardiac development will help combat cardiovascular disorders, among the leading causes of morbidity and mortality worldwide. Among the main mechanisms, we i...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-07-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/15/12251 |
| _version_ | 1797586617066586112 |
|---|---|
| author | Tiziana Angrisano Francesca Varrone Elvira Ragozzino Annalisa Fico Gabriella Minchiotti Mariarita Brancaccio |
| author_facet | Tiziana Angrisano Francesca Varrone Elvira Ragozzino Annalisa Fico Gabriella Minchiotti Mariarita Brancaccio |
| author_sort | Tiziana Angrisano |
| collection | DOAJ |
| description | During cardiac differentiation, numerous factors contribute to the development of the heart. Understanding the molecular mechanisms underlying cardiac development will help combat cardiovascular disorders, among the leading causes of morbidity and mortality worldwide. Among the main mechanisms, we indeed find Cripto. Cripto is found in both the syncytiotrophoblast of ampullary pregnancies and the inner cell mass along the primitive streak as the second epithelial–mesenchymal transformation event occurs to form the mesoderm and the developing myocardium. At the same time, it is now known that cardiac signaling pathways are intimately intertwined with the expression of myomiRNAs, including miR-1. This miR-1 is one of the muscle-specific miRs; aberrant expression of miR-1 plays an essential role in cardiac diseases. Given this scenario, our study aimed to evaluate the inverse correlation between Cripto and miR-1 during heart development. We used in vitro models of the heart, represented by embryoid bodies (EBs) and embryonic carcinoma cell lines derived from an embryo-derived teratocarcinoma in mice (P19 cells), respectively. First, through a luciferase assay, we demonstrated that Cripto is a target of miR-1. Following this result, we observed that as the days of differentiation increased, the Cripto gene expression decreased, while the level of miR-1 increased; furthermore, after silencing miR-1 in P19 cells, there was an increase in Cripto expression. Moreover, inducing damage with a cobra cardiotoxin (CTX) in post-differentiation cells, we noted a decreased miR-1 expression and increased Cripto. Finally, in mouse cardiac biopsies, we observed by monitoring gene expression the distribution of Cripto and miR-1 in the right and left ventricles. These results allowed us to detect an inverse correlation between miR-1 and Cripto that could represent a new pharmacological target for identifying new therapies. |
| first_indexed | 2024-03-11T00:25:39Z |
| format | Article |
| id | doaj.art-9c89bd299f1945e2b7c7b3330dc6cf89 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T00:25:39Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9c89bd299f1945e2b7c7b3330dc6cf892023-11-18T23:02:30ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-07-0124151225110.3390/ijms241512251Cripto Is Targeted by miR-1a-3p in a Mouse Model of Heart DevelopmentTiziana Angrisano0Francesca Varrone1Elvira Ragozzino2Annalisa Fico3Gabriella Minchiotti4Mariarita Brancaccio5Department of Biology, University of Naples Federico II, 80126 Naples, ItalyIRBM S.p.A, 80131 Naples, ItalyDipartimento di Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, 26100 Rome, ItalyStem Cell Fate Laboratory, Institute of Genetics and Biophysics, “A. Buzzati-Traverso”, CNR, 80131 Naples, ItalyStem Cell Fate Laboratory, Institute of Genetics and Biophysics, “A. Buzzati-Traverso”, CNR, 80131 Naples, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131 Naples, ItalyDuring cardiac differentiation, numerous factors contribute to the development of the heart. Understanding the molecular mechanisms underlying cardiac development will help combat cardiovascular disorders, among the leading causes of morbidity and mortality worldwide. Among the main mechanisms, we indeed find Cripto. Cripto is found in both the syncytiotrophoblast of ampullary pregnancies and the inner cell mass along the primitive streak as the second epithelial–mesenchymal transformation event occurs to form the mesoderm and the developing myocardium. At the same time, it is now known that cardiac signaling pathways are intimately intertwined with the expression of myomiRNAs, including miR-1. This miR-1 is one of the muscle-specific miRs; aberrant expression of miR-1 plays an essential role in cardiac diseases. Given this scenario, our study aimed to evaluate the inverse correlation between Cripto and miR-1 during heart development. We used in vitro models of the heart, represented by embryoid bodies (EBs) and embryonic carcinoma cell lines derived from an embryo-derived teratocarcinoma in mice (P19 cells), respectively. First, through a luciferase assay, we demonstrated that Cripto is a target of miR-1. Following this result, we observed that as the days of differentiation increased, the Cripto gene expression decreased, while the level of miR-1 increased; furthermore, after silencing miR-1 in P19 cells, there was an increase in Cripto expression. Moreover, inducing damage with a cobra cardiotoxin (CTX) in post-differentiation cells, we noted a decreased miR-1 expression and increased Cripto. Finally, in mouse cardiac biopsies, we observed by monitoring gene expression the distribution of Cripto and miR-1 in the right and left ventricles. These results allowed us to detect an inverse correlation between miR-1 and Cripto that could represent a new pharmacological target for identifying new therapies.https://www.mdpi.com/1422-0067/24/15/12251CriptomiR-1luciferase assaycardiac differentiationcardiac injury |
| spellingShingle | Tiziana Angrisano Francesca Varrone Elvira Ragozzino Annalisa Fico Gabriella Minchiotti Mariarita Brancaccio Cripto Is Targeted by miR-1a-3p in a Mouse Model of Heart Development International Journal of Molecular Sciences Cripto miR-1 luciferase assay cardiac differentiation cardiac injury |
| title | Cripto Is Targeted by miR-1a-3p in a Mouse Model of Heart Development |
| title_full | Cripto Is Targeted by miR-1a-3p in a Mouse Model of Heart Development |
| title_fullStr | Cripto Is Targeted by miR-1a-3p in a Mouse Model of Heart Development |
| title_full_unstemmed | Cripto Is Targeted by miR-1a-3p in a Mouse Model of Heart Development |
| title_short | Cripto Is Targeted by miR-1a-3p in a Mouse Model of Heart Development |
| title_sort | cripto is targeted by mir 1a 3p in a mouse model of heart development |
| topic | Cripto miR-1 luciferase assay cardiac differentiation cardiac injury |
| url | https://www.mdpi.com/1422-0067/24/15/12251 |
| work_keys_str_mv | AT tizianaangrisano criptoistargetedbymir1a3pinamousemodelofheartdevelopment AT francescavarrone criptoistargetedbymir1a3pinamousemodelofheartdevelopment AT elviraragozzino criptoistargetedbymir1a3pinamousemodelofheartdevelopment AT annalisafico criptoistargetedbymir1a3pinamousemodelofheartdevelopment AT gabriellaminchiotti criptoistargetedbymir1a3pinamousemodelofheartdevelopment AT mariaritabrancaccio criptoistargetedbymir1a3pinamousemodelofheartdevelopment |