Long noncoding RNA H19X is a key mediator of TGF-β–driven fibrosis
Elena Pachera, Shervin Assassi, Gloria A. Salazar, Mara Stellato, Florian Renoux, Adam Wunderlin, Przemyslaw Blyszczuk, Robert Lafyatis, Fina Kurreeman, Jeska de Vries-Bouwstra, Tobias Messemaker, Carol A. Feghali-Bostwick, Gerhard Rogler, Wouter T. van Haaften, Gerard Dijkstra, Fiona Oakley, Maurizio Calcagni, Janine Schniering, Britta Maurer, Jörg H.W. Distler, Gabriela Kania, Mojca Frank-Bertoncelj, Oliver Distler
Elena Pachera, Shervin Assassi, Gloria A. Salazar, Mara Stellato, Florian Renoux, Adam Wunderlin, Przemyslaw Blyszczuk, Robert Lafyatis, Fina Kurreeman, Jeska de Vries-Bouwstra, Tobias Messemaker, Carol A. Feghali-Bostwick, Gerhard Rogler, Wouter T. van Haaften, Gerard Dijkstra, Fiona Oakley, Maurizio Calcagni, Janine Schniering, Britta Maurer, Jörg H.W. Distler, Gabriela Kania, Mojca Frank-Bertoncelj, Oliver Distler
View: Text | PDF
Research Article Autoimmunity Cell biology

Long noncoding RNA H19X is a key mediator of TGF-β–driven fibrosis

Abstract

TGF-β is a master regulator of fibrosis, driving the differentiation of fibroblasts into apoptosis-resistant myofibroblasts and sustaining the production of extracellular matrix (ECM) components. Here, we identified the nuclear long noncoding RNA (lncRNA) H19X as a master regulator of TGF-β–driven tissue fibrosis. H19X was consistently upregulated in a wide variety of human fibrotic tissues and diseases and was strongly induced by TGF-β, particularly in fibroblasts and fibroblast-related cells. Functional experiments following H19X silencing revealed that H19X was an obligatory factor for TGF-β–induced ECM synthesis as well as differentiation and survival of ECM-producing myofibroblasts. We showed that H19X regulates DDIT4L gene expression, specifically interacting with a region upstream of the DDIT4L gene and changing the chromatin accessibility of a DDIT4L enhancer. These events resulted in transcriptional repression of DDIT4L and, in turn, in increased collagen expression and fibrosis. Our results shed light on key effectors of TGF-β–induced ECM remodeling and fibrosis.

Authors

Elena Pachera, Shervin Assassi, Gloria A. Salazar, Mara Stellato, Florian Renoux, Adam Wunderlin, Przemyslaw Blyszczuk, Robert Lafyatis, Fina Kurreeman, Jeska de Vries-Bouwstra, Tobias Messemaker, Carol A. Feghali-Bostwick, Gerhard Rogler, Wouter T. van Haaften, Gerard Dijkstra, Fiona Oakley, Maurizio Calcagni, Janine Schniering, Britta Maurer, Jörg H.W. Distler, Gabriela Kania, Mojca Frank-Bertoncelj, Oliver Distler

×

Figure 8

DDIT4L is a repressor of TGF-β–induced collagen production.

Options: View larger image (or click on image) Download as PowerPoint
DDIT4L is a repressor of TGF-β–induced collagen production. H19X was sil...
H19X was silenced in dermal SSc fibroblasts with 25 nM ASO for a total of 120 hours and stimulated with TGF-β for the last 72 hours in 1% FBS medium. The mRNA and protein levels of DDIT4L were analyzed by qPCR and Western blot respectively. Pictures are representative of n = 5 biological replicates (A). SSc fibroblasts were transfected with 25 nM of ASO and 50 nM of siRNA for 120 hours and stimulated with TGF-β for the last 72 hours in 1% FBS medium. Expression of DDIT4L, H19X, and COL1A1 was analyzed by qPCR (untreated samples) (B). qPCR data were normalized to GAPDH and RPLP0. Data are presented as single values and mean (n = 5–6). Fold change was calculated respective to untreated control set as 1 (dashed line). One-way ANOVA (A and B). *P < 0.05, **P < 0.01, ***P < 0.001.