Transcriptional regulation of cardiac extracellular matrix gene expression and fibroblast phenotype by scleraxis
| dc.contributor.author | Adhikari Bagchi, Rushita | |
| dc.contributor.examiningcommittee | Kardami, Elissavet (Physiology and Pathophysiology) Kirshenbaum, Lorrie (Physiology and Pathophysiology) Dixon, Ian (Physiology and Pathophysiology) Wigle, Jeffrey (Biochemistry and Medical Genetics) Lincoln, Joy (Nationwide Children's Hospital, Columbus, OH) | en_US |
| dc.contributor.supervisor | Czubryt, Michael Paul (Physiology and Pathophysiology) | en_US |
| dc.date.accessioned | 2016-04-18T16:39:14Z | |
| dc.date.available | 2016-04-18T16:39:14Z | |
| dc.date.issued | 2015 | |
| dc.degree.discipline | Physiology and Pathophysiology | en_US |
| dc.degree.level | Doctor of Philosophy (Ph.D.) | en_US |
| dc.description.abstract | Cardiac fibrosis contributes to heart failure by dramatically impairing cardiac function, increasing patient morbidity and mortality. The primary fibrillar collagen expressed in the heart is type I, and increased collagen synthesis is the hallmark of cardiac fibrosis. Our laboratory has shown that the transcription factor scleraxis is sufficient to regulate the gene encoding collagen Iα2. The present thesis identifies and focuses on three key functions of scleraxis in the heart. First, we show that scleraxis is required for production of the cardiac extracellular matrix. Using in vitro and in vivo models, we observed a significant upregulation/reduction of matrix genes in response to induction/loss of scleraxis gene function respectively. In fact, scleraxis overexpression was sufficient to rescue matrix synthesis in scleraxis-null cells. In a murine model of cardiac pressure overload, scleraxis gene deletion blunted the induction of fibrotic collagen gene expression. Second, we provide evidence that scleraxis governs fibroblast-myofibroblast phenotype transition and fibroblast number. Scleraxis gene induction promoted cardiac myofibroblast phenoconversion while knockdown reduced myofibroblast marker gene expression. Scleraxis exerts direct transcriptional control on the a-smooth muscle actin gene-an established marker of myofibroblasts. Scleraxis null mice exhibited a dramatic reduction in cardiac fibroblast numbers- this is attributed to impairment of the epithelial-to-mesenchymal transition program which was marked by a corresponding loss of mesenchymal markers and increased epithelial markers. Loss-of-function experiments using primary cardiac proto-myofibroblasts recapitulated this paradigm, whereas scleraxis gene induction showed a reciprocal effect on mesenchymal markers. Third, data from this study supports the required role of scleraxis in the TGFb/Smad signaling pathway. Scleraxis is strongly upregulated by the potent pro-fibrotic cytokine TGFb, and works synergistically with the canonical Smad signaling pathway to increase Col1a2 expression by cardiac fibroblasts and myofibroblasts. Smad3 induced expression of the fibrillar collagens – an effect that was significantly attenuated following scleraxis knockdown. Smad3 binding to the Col1a2 gene promoter was significantly reduced in scleraxis null hearts. This study involved a comprehensive series of in vitro and in vivo experiments, and is the first to identify scleraxis as a key regulator of multiple fibroblast functions and a potential future target for therapeutic intervention in cardiac fibrosis. | en_US |
| dc.description.note | May 2016 | en_US |
| dc.identifier.uri | http://hdl.handle.net/1993/31243 | |
| dc.language.iso | eng | en_US |
| dc.rights | open access | en_US |
| dc.subject | fibroblast | en_US |
| dc.subject | myofibroblast | en_US |
| dc.subject | transcription | en_US |
| dc.subject | gene regulation | en_US |
| dc.subject | transgenic mice | en_US |
| dc.subject | extracellular matrix | en_US |
| dc.subject | conditional knockout | en_US |
| dc.subject | fibrosis | en_US |
| dc.title | Transcriptional regulation of cardiac extracellular matrix gene expression and fibroblast phenotype by scleraxis | en_US |
| dc.type | doctoral thesis | en_US |