1c211d730ab6330e8044aea474cf27c3 pmc_33333886.pdf 9fe8ec72fb1d362f4625880d79e77e3b3d4266e0 pmc_33333886.pdf 25fe17f95bbd1e4e04d2bfc6011cb944eebb9a6e12398068187ac4ff77e34a71 pmc_33333886.pdf Title: Epigenetic Silencing of Tumor Suppressor miR-124 Directly Supports STAT3 Activation in Cutaneous T-Cell Lymphoma Subject: Increasing evidence supports a potential role for STAT3 as a tumor driver in cutaneous T-cell lymphomas (CTCL). The mechanisms leading to STAT3 activation are not fully understood; however, we recently found that miR-124, a known STAT3 regulator, is robustly silenced in MF tumor-stage and CTCL cells. Objective: We studied here whether deregulation of miR-124 contributes to STAT3 pathway activation in CTCL. Methods: We measured the effect of ectopic mir-124 expression in active phosphorylated STAT3 (p-STAT3) levels and evaluated the transcriptional impact of miR-124-dependent STAT3 pathway regulation by expression microarray analysis. Results: We found that ectopic expression of miR-124 results in massive downregulation of activated STAT3 in different CTCL lines, which resulted in a significant alteration of genetic signatures related with gene transcription and proliferation such as MYC and E2F. Conclusions: Our study highlights the importance of the miR-124/STAT3 axis in CTCL and demonstrates that the STAT3 pathway is regulated through epigenetic mechanisms in these cells. Since deregulated STAT3 signaling has a major impact on CTCL initiation and progression, a better understanding of the molecular basis of the miR-124/STAT3 axis may provide useful information for future personalized therapies. Keywords: cutaneous T-cell lymphoma; mycosis fungoides; Sézary syndrome; STAT3 (signal transducer and activator of transcription-3); JAK/STAT (janus kinase/signal transducer and activator of transcription); miR-124 Author: Lidia García-Colmenero, Jéssica González, Juan Sandoval, Yolanda Guillén, Angel Diaz-Lagares, Evelyn Andrades, Arnau Iglesias, Lara Nonell, Ramon Maria Pujol, Anna Bigas, Lluís Espinosa and Fernando Gallardo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Dec 16 07:32:59 2020 CET ModDate: Wed Dec 16 07:46:09 2020 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 12 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1535635 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NFAGMW+Pxsy Type 1 Builtin yes yes no 10 0 JECVIE+VnURWPalladioL Type 1 Custom yes yes no 14 0 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valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 437 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Study Samples, Cell Lines, and Treatments Destination: subsection.2.1 Item: Title: DNA Methylation Status of the miR-124 Promoter in CTCL Cell Lines and Patients Destination: subsection.2.2 Item: Title: Expression Analysis of miR-124 by qRT PCR Destination: subsection.2.3 Item: Title: Lentiviral Transfection Destination: subsection.2.4 Item: Title: Microarray Expression Profiles Destination: subsection.2.5 Item: Title: Gene Ontology Analysis of Differentially Expressed Genes Destination: subsection.2.6 Item: Title: Ethical Considerations Destination: subsection.2.7 Item: Title: Results Destination: section.3 Children: Item: Title: Silencing of miR-124 as a Consequence of Its Promoter Hypermethylation in Both CTCL Cell Lines and MF Tumor Samples Destination: subsection.3.1 Item: Title: Hypermethylation of the miR-124 Promoter in CTCL Cells Leads to miR-124 Silencing and Increased STAT3 Levels Destination: subsection.3.2 Item: Title: Ectopic miR-124 Expression in CTCL Cells Imposes an Altered Gene Expression Profile Destination: subsection.3.3 Item: Title: CTCL Cells Carrying STAT3 Activation Are Highly Sensitive to Pharmacologic JAK/STAT Inhibition Destination: subsection.3.4 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: section.4 Info: Title: Epigenetic Silencing of Tumor Suppressor miR-124 Directly Supports STAT3 Activation in Cutaneous T-Cell Lymphoma Author: Lidia García-Colmenero, Jéssica González, Juan Sandoval, Yolanda Guillén, Angel Diaz-Lagares, Evelyn Andrades, Arnau Iglesias, Lara Nonell, Ramon Maria Pujol, Anna Bigas, Lluís Espinosa and Fernando Gallardo Subject: Increasing evidence supports a potential role for STAT3 as a tumor driver in cutaneous T-cell lymphomas (CTCL). The mechanisms leading to STAT3 activation are not fully understood; however, we recently found that miR-124, a known STAT3 regulator, is robustly silenced in MF tumor-stage and CTCL cells. Objective: We studied here whether deregulation of miR-124 contributes to STAT3 pathway activation in CTCL. Methods: We measured the effect of ectopic mir-124 expression in active phosphorylated STAT3 (p-STAT3) levels and evaluated the transcriptional impact of miR-124-dependent STAT3 pathway regulation by expression microarray analysis. Results: We found that ectopic expression of miR-124 results in massive downregulation of activated STAT3 in different CTCL lines, which resulted in a significant alteration of genetic signatures related with gene transcription and proliferation such as MYC and E2F. Conclusions: Our study highlights the importance of the miR-124/STAT3 axis in CTCL and demonstrates that the STAT3 pathway is regulated through epigenetic mechanisms in these cells. Since deregulated STAT3 signaling has a major impact on CTCL initiation and progression, a better understanding of the molecular basis of the miR-124/STAT3 axis may provide useful information for future personalized therapies. 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