c44b6d9fa9dfac50601c0bd5507e8f5e pmc_35215234.pdf ff75c23203f3cbb7c39def08a4cc2c0d15f55237 pmc_35215234.pdf ac9458aed94e90a53bf475f7a72de5ed5efd0e1c2693760cd2bde06a06a89da0 pmc_35215234.pdf Title: Epigenetic Modulation of Gremlin-1/NOTCH Pathwayin Experimental Crescentic Immune- Mediated Glomerulonephritis Subject: Crescentic glomerulonephritis is a devastating autoimmune disease that without early and properly treatment may rapidly progress to end-stage renal disease and death. Current immunosuppressive treatment provides limited efficacy and an important burden of adverse events. Epigenetic drugs are a source of novel therapeutic tools. Among them, bromodomain and extraterminal domain (BET) inhibitors (iBETs) block the interaction between bromodomains and acetylated proteins, including histones and transcription factors. iBETs have demonstrated protective effects on malignancy, inflammatory disorders and experimental kidney disease. Recently, Gremlin-1 was proposed as a urinary biomarker of disease progression in human anti-neutrophil cytoplasmic antibody (ANCA)-associated crescentic glomerulonephritis. We have now evaluated whether iBETs could regulate Gremlin-1 in experimental anti-glomerular basement membrane nephritis induced by nephrotoxic serum (NTS) in mice, a model resembling human crescentic glomerulonephritis. In NTS-injected mice, the iBET JQ1 inhibited renal Gremlin-1 overexpression and diminished glomerular damage, restoring podocyte numbers. Chromatin immunoprecipitation assay demonstrated BRD4 enrichment of the Grem-1 gene promoter in injured kidneys, consistent with Gremlin-1 epigenetic regulation. Moreover, JQ1 blocked BRD4 binding and inhibited Grem-1 gene transcription. The beneficial effect of iBETs was also mediated by modulation of NOTCH pathway. JQ1 inhibited the gene expression of the NOTCH effectors Hes-1 and Hey-1 in NTS-injured kidneys. Our results further support the role for epigenetic drugs, such as iBETs, in the treatment of rapidly progressive crescentic glomerulonephritis. Keywords: crescentic glomerulonephritis; chronic kidney disease; bromodomain; BET proteins; Gremlin; NOTCH Author: Lucia Tejedor-Santamaria, Jose Luis Morgado-Pascual, Laura Marquez-Exposito, Beatriz Suarez-Alvarez, Raul R. Rodrigues-Diez, Antonio Tejera-Muñoz, Vanessa Marchant, Sergio Mezzano, Carlos Lopez-Larrea, Anna Sola, Gema Maria Fernandez-Juarez, Alberto Ortiz, Sandra Rayego-Mateos and Marta Ruiz-Ortega Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Jan 21 02:01:24 2022 CET ModDate: Fri Jan 21 02:04:32 2022 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2723636 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SWKKKB+VnURWPalladioL Type 1 Custom yes yes no 10 0 PHCWYT+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 CKIBBL+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 FCXLSV+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 32 0 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Nephrotoxic Nephritis Destination: subsection.2.1 Item: Title: Gremlin-1 Is Overexpressed in Experimental Nephrotoxic Nephritis in Mice: Effect of BET Inhibition Destination: subsection.2.2 Item: Title: Gremlin-1 Is One of the Specific Targets of BET Inhibition Destination: subsection.2.3 Item: Title: JQ1 Inhibits the NOTCH Pathway in Experimental Nephrotoxic Nephritis Destination: subsection.2.4 Item: Title: Differential Gene Expression in Experimental Nephrotoxic Nephritis: Impact of BET Inhibition Destination: subsection.2.5 Item: Title: BET Inhibition Diminished the Renal Expression of the Chemokine Ccl-8 in Experimental Anti-glomerular Basement Membrane Nephritis Destination: subsection.2.6 Item: Title: BET Inhibition Downregulates Gremlin-1 Expression in Other Models of Renal Fibrosis Destination: subsection.2.7 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Ethics Statement Destination: subsection.4.1 Item: Title: Experimental Models Destination: subsection.4.2 Item: Title: Histology and Immunohistochemistry Destination: subsection.4.3 Item: Title: Gene Expression Studies Destination: subsection.4.4 Item: Title: Protein Studies Destination: subsection.4.5 Item: Title: Chromatin Immunoprecipitation Destination: subsection.4.6 Item: Title: Statistical Analysis Destination: subsection.4.7 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: Epigenetic Modulation of Gremlin-1/NOTCH Pathwayin Experimental Crescentic Immune- Mediated Glomerulonephritis Author: Lucia Tejedor-Santamaria, Jose Luis Morgado-Pascual, Laura Marquez-Exposito, Beatriz Suarez-Alvarez, Raul R. Rodrigues-Diez, Antonio Tejera-Muñoz, Vanessa Marchant, Sergio Mezzano, Carlos Lopez-Larrea, Anna Sola, Gema Maria Fernandez-Juarez, Alberto Ortiz, Sandra Rayego-Mateos and Marta Ruiz-Ortega Subject: Crescentic glomerulonephritis is a devastating autoimmune disease that without early and properly treatment may rapidly progress to end-stage renal disease and death. Current immunosuppressive treatment provides limited efficacy and an important burden of adverse events. Epigenetic drugs are a source of novel therapeutic tools. Among them, bromodomain and extraterminal domain (BET) inhibitors (iBETs) block the interaction between bromodomains and acetylated proteins, including histones and transcription factors. iBETs have demonstrated protective effects on malignancy, inflammatory disorders and experimental kidney disease. Recently, Gremlin-1 was proposed as a urinary biomarker of disease progression in human anti-neutrophil cytoplasmic antibody (ANCA)-associated crescentic glomerulonephritis. We have now evaluated whether iBETs could regulate Gremlin-1 in experimental anti-glomerular basement membrane nephritis induced by nephrotoxic serum (NTS) in mice, a model resembling human crescentic glomerulonephritis. In NTS-injected mice, the iBET JQ1 inhibited renal Gremlin-1 overexpression and diminished glomerular damage, restoring podocyte numbers. Chromatin immunoprecipitation assay demonstrated BRD4 enrichment of the Grem-1 gene promoter in injured kidneys, consistent with Gremlin-1 epigenetic regulation. Moreover, JQ1 blocked BRD4 binding and inhibited Grem-1 gene transcription. The beneficial effect of iBETs was also mediated by modulation of NOTCH pathway. JQ1 inhibited the gene expression of the NOTCH effectors Hes-1 and Hey-1 in NTS-injured kidneys. Our results further support the role for epigenetic drugs, such as iBETs, in the treatment of rapidly progressive crescentic glomerulonephritis. 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410, 468 Annotation: Subtype: Link Rect: 370, 422, 408, 433 Annotation: Subtype: Link Rect: 414, 422, 450, 433 Annotation: Subtype: Link Rect: 353, 389, 391, 399 Annotation: Subtype: Link Rect: 397, 389, 433, 399 Annotation: Subtype: Link Rect: 95, 354, 133, 364 Checksum: b6b024ba Type: CRC32 Checksum: c44b6d9fa9dfac50601c0bd5507e8f5e Type: MD5 Checksum: ff75c23203f3cbb7c39def08a4cc2c0d15f55237 Type: SHA-1 Checksum: ac9458aed94e90a53bf475f7a72de5ed5efd0e1c2693760cd2bde06a06a89da0 Type: SHA-256