Epigenetics and Cancer Ya-Hui Chi March 24, 2016 1 Institute of Biotechnology and Pharmaceutical Research, NHRI, Zhunan, Taiwan Question 1: What causes cancer? Douglas Hanahan and Robert A. Weinberg 2 1
The Genetic Code and the Epigenetic Code 3 Stem cell therapy Self-renewal vs. Pluripotency Question 1: Why there are different cell types in an organism with identical gene sequence? ES cells 4 2
Changes in genetic sequence during aging? Question 2: Do identical twins have different susceptibility to cancer and other diseases? 5 W. LOVELACE/GETTY British gangster Ronnie Kray (right) was bisexual, unlike his twin, Reggie (left). The disparity could be down to epigenetics. http://www.nature.com/nature/journal/v487/n7407/full/487298a.html?wt.ec_id=nature-20120719 3
Cancer susceptibility among twins Brest cancer Hereditary 29% Mendelian diseases vs. Ovarian cancer Non-Mendelian diseases Hereditary 22% 7 The double helix and the nucleosome The double helix http://www.mpi-inf.mpg.de/~cbock/privat/overview.htm 8 4
Histones and the epigenetic code (Image: Nature 441, 143-145 (11 May 2006) 9 Epigenetic modification of core histones Epigenetic modifications: Me: Histone methylation Ac: Histone acetylation P: Histone phosphorylation SU: Histone sumoylation Ub: Histone ubiquitination DNA methylation 10 5
Nature Reviews Drug Discovery 8, (September 2009) doi:10.1038/nrd2974 2016/3/22 Histone and the epigenetic modification enzymes 724-732 Acetylation: histone acetyltransferases and deacetylases. Methylation: histone methyltransferases and demethylases, DNA methyltransferases. DNA demethylase? Phosphorylation: kinases. Ubiquitination: Ub ligases. Nature Reviews Drug Discovery 8, 724-732 (September 2009) 11 6
Nature Reviews Drug Discovery 8, (September 2009) doi:10.1038/nrd2974 2016/3/22 Histone and the epigenetic modification enzymes 724-732 https://www.youtube.com/watch?v=tj_6dcutrnm 13 Micro RNA http://cardiovascres.oxfordjournals.org/content/79/4/553 7
Long non-coding RNA http://www.bloodjournal.org/content/121/24/4842 Mechanisms of LncRNA action. In numerous tissues, lncrnas (indicated in green) have been shown to regulate gene expression at multiple levels: chromatin, transcription, mrna, translation, and protein. Hematopoietic lncrnas may act at any of these levels. MicroRNA sponge refers to the ability of lncrnas to sequester cellular micrornas and prevent them from binding mrna targets. Chromosome regions with differential DNA methylation in young and old monozygous twins Significant DNA methylation changes are indicated as thick red and green blocks in the ideograms. The 50-year-old twin pair shows abundant changes in the pattern of DNA methylation (green=hypermethylation and red=hypomethylation), 3-year-old twins have a very similar DNA methylation (yellow). (Image Copyright (2005) National 16 Academy of Sciences, U.S.A) 8
Histone acetyltransferase (HAT) and histone deacetylase (HDAC) Histone acetyltransferase Histone deacetylase GNAT family GCN5 PCAF HAT1 MYST family MYST1 MYST2 HTATIP p300/cbp family p300 CBP Nuclear receptor coactivators AIB1 General transcription factors TAFII250 Class I HDAC1 HDAC2 HDAC3 HDAC8 Class IIa HDAC4 HDAC5 HDAC7 HDAC9 Class IIb HDAC6 HDAC10 Class III SIRT1 SIRT2 SIRT3 SIRT4 SIRT5 SIRT6 SIRT7 Class IV HDAC11 17 Histone methyltransferase (HMT), histone demethylase and DNA methyltransferase (DNMT) Histone methyltransferase H3K4 MLL SET1 SMYD3 H3K9 SUV39H1 SUV39H2 G9a EuHMT SETDB1/ESET RIZ1 H3K27 EZH2 H3K36 NSD1 SMYD2 Histone demethylase H3K4 LSD1 SMCX / JARID1C SMCY / JARID1D RBP2 / JARID1A PLU1/JARID1B H3K9 JHDM2A (JMJD1A) JMJD2A JMJD2B JMJD2C JMJD2D H3K27 UTX JMJD3 H3K36 JMJD2A JMJD2B JMJD2C FBXL10 / JHDM1B DNA methyltransferase DNMT1 DNMT2 DNMT3A DNMT3B DNMT3L 18 9
The histone code hypothesis Strahl BD et al. (2000) Nature 19 Histone Modifying Enzymes Outcome of Gene Expression There are many examples in our everyday lives where common inputs result in differential outcomes, depending on specific control mechanisms. Consider, for example, how a talented musician can produce a wide variety of music on a guitar. A particular piece of music is encoded by the sequence and timing of notes, that are typically transcribed onto sheet music. The guitarist translates the sheet music by spatial and temporal modifications to the guitar strings; that is, he/ she holds down different strings, at different frets, at different times to produce the music (Figure 2A/B). Figure 2 A/B - (A) Music is encoded in the form of notations on sheet music, that define the notes to be played and the tempo at which they are played. (B) A guitarist translates the sheet music into audible music by spatial and temporal control of the guitar strings. In a like manner, gene transcription is spatially and temporally controlled by epigenetic modifications, catalyzed by specific enzymes. http://www.epizyme.com/epigentics/histone-modifying-enzymes.asp 10
Dynamic structure of the chromatin in a cycling cell http://adasilva.glogster.com/chapter-10-bio/ 21 http://www.biology-online.org/biology-forum/viewtopic.php?f=15&t=18785 Coordinated recruitment of histone-modifying activities Rosenfeld MG et al. (2006) Gene Dev. 22 11
Nuclear morphology of undifferentiated and differentiated cells undifferentiated (ESC) differentiated Niwa H. (2007) Development Question 3: What are the differences between undifferentiated and differentiated cells in nuclear morphology? 23 Epigenetic Modification and Transcriptional Control Closed chromatin: transcriptionally repressive Open chromatin: transcriptionally active Kristeleit (2004) Expert Opin. Emerg. Drugs 24 12
Question 6: Which nucleotide can be methylated? S-adenosylmethionine (SAM) : methylation intermediates 25 DNA demethylation TET: ten eleven translocation TDG: thymine-dna glycosylase C S Nabel, R M Kohli Science 2011;333:1229-1230 Published by AAAS 26 13
DNA methylation-induced histone modification for gene silencing http://www.ncc.go.jp/en/nccri/divisions/14carc/14carc01.html 27 Analysis of DNA methylation using bisulphite sequencing http://www.methylogix.com/genetics/protocols.shtml-dateien/schumachersguide1.html http://www.epigenomics.de/en/science-and-technologies/bisulfite-technology/ 28 14
Histone Acetylation 29 Acetylation and deacetylation of the lysine residue 30 15
Timeline of histone acetylation discovery Lee K. (2007) Nat. Rev. Mol. Cell Bio. 31 Histone acetylation and transcriptional control 32 16
Epigenetic gene activation http://www.nature.com/bjc/journal/v90/n4/fig_tab/6601575f2.html S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH)] : methylation intermediates 33 Epigenetic gene silencing http://www.nature.com/bjc/journal/v90/n4/fig_tab/6601575f2.html 34 17
Histone methylation and transcriptional control Niwa H. (2007) Development 35 Epigenetics, Development and Cell Fate Commitment 36 18
Epigenetic gene regulation during mammalian development Reik (2007) Nature 37 Phenotypes of mice knocked out for histone modification enzymes Haberland M. (2009) Nat Rev Genetics 38 19
Huang S. (2009) BioEssays 39 Pluripotency and the transcriptional regulatory circuitry Jaenisch R. and Young R. (2009) Cell 40 20
Nuclear reprogramming genetic code vs. epigenetic code Yamanaga S. (2010) Nature 41 Dolly the sheep http://science.howstuffworks.com/genetic-science/cloning3.htm 42 21
Different methods of reprogramming require transcriptional regulators and passage through cell division 43 Egli D. (2008) Nat. Rev. Mol. Cell. Bio. 44 22
45 Bisulfite genomic sequencing of the promoter regions of OCT3/4, REX1, and NANOG in ips cells ips NTERA-2: a human embryonic carcinoma cell line HDF: human dermal fibroblasts Takahashi K. (2007) Cell 46 23
The epigenetic mark of embryonic stem cells 47 Epigenetic modification inhibitors enhanced generation efficiency of ips NPC MEFs BIX: Histone methyltransferate G9a inhibitor BayK: a L-channel calcium agonist RG108: a DNMT (DNA methyltransferase) inhibitor 48 24
Epigenetic modification inhibitors enhanced generation efficiency of ips + 4 factors 5 -azacytidine (5 -azac): a DNA methyltransferase inhibitor Dexamethasone (dex): a synthetic glucocorticoid, anti-inflammation Valproic acid (VPA): a histone deacetylase inhibitor SAHA: a histone deacetylase inhibitor TSA: a histone deacetylase inhibitor 49 CM+FGF2 SB: sodium butyrate, a HDAC inhibitor +SB, no FGF2 CM+FGF2 +SB no FGF2 With feeder No feeder; on Matrigel 50 25
51 Epigenetics and Cancer 52 26
Six hallmarks of cancer Epigenetic silencing of tumor suppressor genes (examples in parentheses) plays a significant role in the development of each of these six tumor traits http://www.bioscience.org/2005/v10/af/1668/figures.htm 53 Epigenetic targets in cancer http://epi.grants.cancer.gov/bulletins/nov1904.html 54 27
Interaction of TF and epigenetic modification factors Two possible mechanisms by which epigenetic modification can lead to cancer. (A) A previously unmethylated tumor suppressor gene such as p53 becomes methylated. Transcription factor(s) (TF) can no longer bind the promoter region, the gene is not expressed, and damaged cells are allowed to proliferate and become cancerous. (B) A protooncogene is demethylated, allowing TFs to initiate transcription and express the protein product. As in (A), uncontrolled cell growth ensues and leads to cancer. Copyright 2008 Nature Education. http://www.nature.com/scitable/content/two-possible-mechanisms-by-which-epigenetic-modification-24705 55 DNA methylation, gene expression and cancer DNA methylation is normally associated with gene silencing http://www1.elsevier.com/homepage/sab/oncoserve/cl_mr/nephew.htm 56 28
DNA methylation, gene expression and cancer DNA hypomethylation and aberrant gene activation in cancer http://www.deduveinstitute.be/dna_hypomethylation.php 57 DNA methylation, gene expression and cancer DNA hypermethylation and aberrant gene silencing in cancer http://www1.elsevier.com/homepage/sab/oncoserve/cl_mr/nephew.htm 58 29
Histone acetylation, Figure 1 Epigenetic gene gene expression silencing in cancer. and cancer Histone acetylation is normally associated with gene activation Yoo CB and Jones PA (2006) Epigenetic therapy of cancer: past, present and future Nature Reviews Drug Discovery 5: 37 50 doi:10.1038/nrd1930 59 The epigenetic progenitor model of cancer tumor-suppressor gene (TSG) tumor-progenitor genes (TPG) gatekeeper mutation (GKM) oncogene (ONC) Feinberg AP et al. (2005) The epigenetic progenitor origin of human cancer Nat Rev gene. 7: 21 33 doi:10.1038/nri1748 60 30
The epigenetic progenitor model of cancer crypt paneth 61 Age-dependent DNA methylation and cancer 62 31
Epigenetic Therapy 63 Epizyme http://www.youtube.com/v/8bwdhdwfpu 8?autoplay=1&rel=0&enablejsapi=1&playe rapiid=ytplayer 64 32
Therapeutic targeting of the hallmarks of cancer Douglas Hanahan and Robert A. Weinberg 65 From Epigenomics: Roadmap for regulation 518, 314 316 (19 February 2015) doi:10.1038/518314a The Roadmap Epigenomics Project has produced reference epigenomes that provide information on key functional elements controlling gene expression in 127 human tissues and cell types 2, 3, 4, 5, 6, 7, 8, 9, and encompassing embryonic and adult tissues, from healthy individuals and those with disease. a, Many of the adult tissues investigated were broken down by cell type or region blood into several types of immune cell, for instance, and the brain into regions including the hippocampus and dorsolateral prefrontal cortex. Tissue samples and cells were subjected to a range of epigenomic analyses, along with genome sequencing and genome-wide association studies (GWAS). b, Embryonic stem (ES) cells, which are taken from the embryo at the 'blastocyst' stage and can give rise to almost every cell type in the body, were used to analyse, for example, the differentiation of stem cells into different neuronal lineages. The ES-cell-derived cell lines underwent the same epigenomic analyses as the tissue samples. 33
318 NATURE VOL 518 19 FEBRUARY 2015 Data sets available for each reference epigenome 34
Hypermethylated genes commonly used for cancer detection Carins P. (2007) Nat. Rev. Cancer 69 70 35
72 Haberland M. (2009) Nat Rev Genetics 36
Hypermethylated genes commonly used for cancer detection Carins P. (2007) Nat. Rev. Cancer 73 HDAC inhibitors for cancer therapy 74 37
HDAC inhibitors for cancer therapy Kristensen L. (2009) Eur. J. Pharm. 75 DNMT inhibitors for cancer therapy Kristensen L. (2009) Eur. J. Pharm. 76 38
H2AX inducers for cancer therapy Bonner W. (2008) Nat. Rev. Cancer 77 Tumour-selective action of HDACi in acute promyelocytic leukaemia http://www.medscape.com/viewarticle/521403_8 78 39
Discussion 79 80 40
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