کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5589337 | 1569827 | 2017 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Profiling of human epigenetic regulators using a semi-automated real-time qPCR platform validated by next generation sequencing
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کلمات کلیدی
HDAC7HDAC4HDAC9WDR5ALPLHDAC2Hdac5UTYHDAC1histone deacetylase 9CBXEpigenetic regulatorHDAC11Epigenetic regulatorsBRDKMTHDACMSCRT-qPCRMesenchymal stromal/stem cellRNA-seqKDMAlkaline phosphatase - آلکالین فسفاتاز یا فسفاتاز قلیاییEpigenetics - اپی ژنتیکBromodomain - برومودومینRNA sequencing - ترتیب RNAdeacetylase - دیاستیلازMesenchymal stem cell - سلول های بنیادی مزانشیمیlysine demethylase - لیزین دیمی ایزاlysine methyltransferase - لیزین متیل ترانسفرازMethyltransferase - متیل ترانسفرازHistone - هیستونhistone acetyl transferase - هیستون استیل ترانسفرازhistone deacetylase - هیستون داستیلازhistone deacetylase 1 - هیستون دیازتیلاز 1Histone deacetylase 2 - هیستون دیازتیلاز 2Histone deacetylase 5 - هیستون دیازتیلاز 5Histone deacetylase 4 - هیستون دیازکتیلاز 4Panobinostat - پانوبینواستاتHAT - کلاه
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Epigenetic mechanisms control phenotypic commitment of mesenchymal stromal/stem cells (MSCs) into osteogenic, chondrogenic or adipogenic lineages. To investigate enzymes and chromatin binding proteins controlling the epigenome, we developed a hybrid expression screening strategy that combines semi-automated real-time qPCR (RT-qPCR), next generation RNA sequencing (RNA-seq), and a novel data management application (FileMerge). This strategy was used to interrogate expression of a large cohort (n > 300) of human epigenetic regulators (EpiRegs) that generate, interpret and/or edit the histone code. We find that EpiRegs with similar enzymatic functions are variably expressed and specific isoforms dominate over others in human MSCs. This principle is exemplified by analysis of key histone acetyl transferases (HATs) and deacetylases (HDACs), H3 lysine methyltransferases (e.g., EHMTs) and demethylases (KDMs), as well as bromodomain (BRDs) and chromobox (CBX) proteins. Our results show gender-specific expression of H3 lysine 9 [H3K9] demethylases (e.g., KDM5D and UTY) as expected and upregulation of distinct EpiRegs (n > 30) during osteogenic differentiation of MSCs (e.g., HDAC5 and HDAC7). The functional significance of HDACs in osteogenic lineage commitment of MSCs was functionally validated using panobinostat (LBH-589). This pan-deacetylase inhibitor suppresses osteoblastic differentiation as evidenced by reductions in bone-specific mRNA markers (e.g., ALPL), alkaline phosphatase activity and calcium deposition (i.e., Alizarin Red staining). Thus, our RT-qPCR platform identifies candidate EpiRegs by expression screening, predicts biological outcomes of their corresponding inhibitors, and enables manipulation of the human epigenome using molecular or pharmacological approaches to control stem cell differentiation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 609, 20 April 2017, Pages 28-37
Journal: Gene - Volume 609, 20 April 2017, Pages 28-37
نویسندگان
Amel Dudakovic, Martina Gluscevic, Christopher R. Paradise, Halil Dudakovic, Farzaneh Khani, Roman Thaler, Farah S. Ahmed, Xiaodong Li, Allan B. Dietz, Gary S. Stein, Martin A. Montecino, David R. Deyle, Jennifer J. Westendorf, Andre J. van Wijnen,