title: Prolonged re-expression of the hypermethylated gene EPB41L3 using artificial transcription factors and epigenetic drugs.
abstract: Epigenetic silencing of tumor suppressor genes (TSGs) is considered a significant event in the progression of cancer. For example, EPB41L3, a potential biomarker in cervical cancer, is often silenced by cancer-specific promoter methylation. Artificial transcription factors (ATFs) are unique tools to re-express such silenced TSGs to functional levels; however, the induced effects are considered transient. Here, we aimed to improve the efficiency and sustainability of gene re-expression using engineered zinc fingers fused to VP64 (ZF-ATFs) or DNA methylation modifiers (ZF-Tet2 or ZF-TDG) and/or by co-treatment with epigenetic drugs [5-aza-2'-deoxycytidine or Trichostatin A (TSA)]. The EPB41L3-ZF effectively bound its methylated endogenous locus, as also confirmed by ChIP-seq. ZF-ATFs reactivated the epigenetically silenced target gene EPB41L3 (∼ 10-fold) in breast, ovarian, and cervical cancer cell lines. Prolonged high levels of EPB41L3 (∼ 150-fold) induction could be achieved by short-term co-treatment with epigenetic drugs. Interestingly, for otherwise ineffective ZF-Tet2 or ZF-TDG treatments, TSA facilitated re-expression of EPB41L3 up to twofold. ATF-mediated re-expression demonstrated a tumor suppressive role for EPB41L3 in cervical cancer cell lines. In conclusion, epigenetic reprogramming provides a novel way to improve sustainability of re-expression of epigenetically silenced promoters.
authors.affiliation: a Department of Pathology and Medical Biology.
authors.forename: Christian
authors.initials: C
authors.lastname: Huisman
authors.forename: Monique G P
authors.initials: MG
authors.lastname: van der Wijst
authors.forename: Fahimeh
authors.initials: F
authors.lastname: Falahi
authors.forename: Juul
authors.initials: J
authors.lastname: Overkamp
authors.forename: Gellért
authors.initials: G
authors.lastname: Karsten
authors.forename: Martijn M
authors.initials: MM
authors.lastname: Terpstra
authors.forename: Klaas
authors.initials: K
authors.lastname: Kok
authors.forename: Ate G J
authors.initials: AG
authors.lastname: van der Zee
authors.forename: Ed
authors.initials: E
authors.lastname: Schuuring
authors.forename: G Bea A
authors.initials: GB
authors.lastname: Wisman
authors.forename: Marianne G
authors.initials: MG
authors.lastname: Rots
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D045744:Cell Line, Tumor; D019175:DNA Methylation / Q000187:drug effects*; D000077209:Decitabine; D044127:Epigenesis, Genetic; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D042421:Histone Code / Q000187:drug effects; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008840:Microfilament Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; C116887:EPB41L3 protein, human; D006877:Hydroxamic Acids; D008840:Microfilament Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; C012589:trichostatin A; D000077209:Decitabine; D006655:Histone Deacetylases; D001374:Azacitidine
keywords: DNA demethylases; anti-cancer drug response; artificial transcription factors; epigenome editing; methylation biomarkers; novel antitumor agents
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