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
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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

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Figure 7

H19X targets DDIT4L via 3D genomic interaction and chromatin rearrangements.

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H19X targets DDIT4L via 3D genomic interaction and chromatin rearrangeme...
BJ5TA cells were stimulated with TGF-β for 6 hours in 1% FBS medium. ChIRP peaks for H19X capture are shown as fold enrichment relative to input for 2 sets of probes (even, odd, red tracks). ChIRP tracks for H3K27ac (light blue) and H3K4me1 (dark blue) were obtained from publicly available data sets (57). ATAC-Seq tracks showing variations in chromatin accessibility after H19X silencing. H19X was silenced in dermal SSc fibroblasts with 25 nM ASO (light green) and compared with SSc fibroblasts transfected with scrambled control (dark green) (A). Hi-C data (58) obtained from publicly available data sets revealed interactions between H19X binding site, the active enhancer, and DDIT4L promoter. Query regions: DDIT4L promoter, DDIT4L enhancer (chr4:100294478), H19X binding site (chr4:100375842). Peak signals indicate interaction events and are highlighted by red circles (B). Schematic representation of DDIT4L gene (blue) and its regulatory elements: DDIT4L enhancer (green, in light green it is highlighted where H19X silencing caused variation in chromatin accessibility) and H19X binding site (red) (C).