epigenetics: ”from phenomenon to field” and impact on plastic surgery research basic science...
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Epigenetics:”From Phenomenon to Field”
and Impact on Plastic Surgery Research
Basic Science Seminar SeriesUniversity of MichiganDepartment of Surgery
Section of Plastic Surgery
Brent M. Egeland, MDOctober 13, 2007
Outline
Importance History Definitions Chemistry Laboratory Techniques Implications in Plastic Surgery
Why?
Definition: Epigenetics
Waddington 1942 Originally a combination of the
words "genetics" and "epigenesis" Epigenesis: differentiation of cells
from a totipotent state in embryonic development (used in contrast to "preformationism").
Definition: Epigenetics
69th CSH Symposia (2004): Conference Dedicated to Epigenetics No definition could be agreed upon
”An epigenetic phenomenon is a switch-like change in phenotype that is heritable, but does not involve DNA mutation.” C. David Allis
Why Epigenetics? Post-Genomic Era
ENCODE Genomics and proteomics is not enough Contol of above beyond promoters and
regulators is of utmost importance in: Oncogenesis Stem Cell Biology Genetic therapy Aging
History
Coalesced into field of study that affects all areas of biology
Collection of apparently disparate phenomena Over 1000 manuscripts on topic First observations dates to 1941
HJ Mueller ”eversporting displacement” McClintock noted that these positional effects were
the basis of ”mutible loci”
History
X-Chromosome Inactivation Pre-32 cell stage mosaic cell population
History
1980's: Molecular Biology and Genetics
1990's: Genomics 2003: Human Genome
Complete 2004: ENCODE 2005: NCI announce:
Human Epigenome Project
Definition: Epigenome
”Sum of genome-wide epigenetic patterns, distinguishes and defines one tissue from another, stem cells from somatic cells, and aged from young cells.”
National Cancer Institute (NCI), Division of Cancer Biology (DCB)
Workshop on Defining the Epigenome: Addressing the Value and Scope of a Human Epigenome Project
November 28–29, 2005Andrew P. Feinberg and Peter A. Jones
Chromatin
Euchromatin: Uncompacted Active
Heterochromatin: Compacted Untranscribed Changes with cell cycle
30nm Fiber
Beads on a String
H1: Linker histone: contacts the exit/entry of the DNA strand on the nucleosome.
Linker DNA between Histones
Nucleosome
Histone
DNA Packaging
Varys greatly with Cell Cycle DNA never completely ”naked”
Scaffold or Dynamic?
Is chromatin simply a scaffold for packaging DNA?
Dynamic
Histones are Modified DNA is Modified Heritable transcription states
Histone Modifications
K K
Acetylation = Transcriptional CompetenceMethylation = Transcriptionally Silenced
DNA Cellular Differentiation
All cells inherit the same DNA sequences Cellular differentiation processes rely strongly on
epigenetic rather than genetic inheritance.
DNA Modification
Chemical modification of DNA Can be inherited without sequence
change Part of the epigenetic code Methylation is most characterized
epigenetic mechanism Commonly occurs at ”junk” DNA
DNA Methylation
DNA Methylation
CpG Islands
Laboratory Techniques
Chromatin Immunoprecipitation Inhibition of Deacetylases (HDAC-I) Site-specific analysis of histone
methylation Multigenerational assesment of above Methylation-specific PCR in-situ
Hybridization Q-PCR in Combination with ChIP
Consequences
Genome is static Genotype Germ Cell Inheritance
Epigenome is dynamic Phenotype Somatic epigenetic inheritance
Cellular "memory" Most terminally (irreversibly)
differentiate Morphogens activate/silence
genes – DNA is not altered
Consequences Mammals cannot respond to loss of some tissues
Inability to regenerate limbs
Plant cells do not terminally differentiate
Evolution
Short-term adaptation allowing for reversible phenotype
Allows multigenerational revisible phenotypic switch to express and repress that particular gene
DNA message is unchanged Differential mutation rates associated
with epigenetic features to control the mutation rates of particular genes
Teratogens
Exert effects on fetus by epigenetic means Epigenetic effects may preserve effect of
teratogen throughout life of child Can a teratogen exert influence offspring?
Diethylstilbestrol: Second and possibly Third generation effects
Research
2000 2001 2002 2003 2004 2005 2006 2007
0
20
40
60
80
100
120
140
160
Epigenetic Grants
Year
Gra
nts
Aw
ard
ed
Why Do I Care?
?
Plastic Surgery
Fundamentally involved in clinical epigenetics Aging Oncology Wound Healing
Basic Science in Plastic Surgery Regenerative medicine Stem Cell biology Organogenesis Neural plasticity
Oncology
Clin Cancer Res 2007;13(13) July 1, 2007
Aging Chronological or Epigenetically
Defined? Free-Radicals Oxidants Telomeres methylation
Aging
Change in phenotype with methylation of DNA.
Werner Syndrome
Breast Cancer
Stem Cells
Organogenesis
Neuroal Plasticity
Why does the CNS lack plasticity?
What are the possibilitys of increasing this plasticity towards regenerative capacity?
Hernia
Male Pattern Baldness
The list goes on....
Are there differing methylation signatures of fibroblasts in normal, hypertrophic, keloid scarring.
Can fibrotic tissue (fibroblasts) be reversed to form skin, bone, muscle?
Is cleft lip epigenetically inhereited?
Can we Modify Epigenetics?
Can methylation be reversed? Rate decreased?
Environment? Diet? Pharmacologically?
Diet
Agouti Gene: Yellow fur, obesity, diabetes, tumorogenesis
.
Methyl Donor Diet: (onions, garlic, beets, folic acid, B12, genestein)
No Supplements
HCC: Histone deacetylase inhibitors (HDAC-I) & Demethylating agents
Journal of Clinical Oncology, 2006 ASCO Annual Meeting Proceedings Part I. Vol 24, No. 18S (June 20 Supplement), 2006: 10066
Decitabine & Azacitidine
Mechanism of action: Phosphorylation and direct insertion into
DNA Blocks effects of DNA methyltransferase Causing hypomethylation fo DNA. May reverse inactivation of supressor
genes Can lead to cytotoxicity, bone marrow
suppression
Azacitidine
aza-
5-aza-2'-deoxycytidine
References
Age-related DNA methylation changes in normal huma...[Clin Cancer Res. 2007] - PubMed Result
A method to study the expression of DNA methyltran...[Methods Mol Biol. 2007] - PubMed Result
Clinical Trial: Epigenetics in the Aging Process
Decreased androgen receptor gene methylation in pr...[J Clin Endocrinol Metab. 2006] - PubMed Result
EMF Award: Epigenetic Control of Senescence and Aging
Entirely new approach to the treatment of aging
Epidermal stem cells are defined by global histone...[PLoS ONE. 2007] - PubMed Result
Epigenetic inactivation of the premature aging Werner syndrome gene in human cancer -- Agrelo et al. 103 (23): 8822 -- Proceedings of the National Academy of Sciences
From The Cover: Epigenetic differences arise during the lifetime of monozygotic twins -- Fraga et al. 102 (30): 10604 -- Proceedings of the National Academy of Sciences
Google Image Result for http://edoc.hu-berlin.de/dissertationen/seitz-stefanie-2004-10-20/HTML/seitz_html_21996ee1.png
Google Image Result for http://www.biophysik.uni-bremen.de/radmacher/gallery/osteoblast_b.gif
Google Image Result for http://www.med.ufl.edu/biochem/keithr/fig2pt2_flowchart-sm.gif
Google Image Result for http://www.mun.ca/biology/desmid/brian/BIOL2250/Week_Two/bead2sol.jpg
Human hair genealogies and stem cell latency. [BMC Biol. 2006] - PubMed Result
IngentaConnect Techniques Used in Studies of Age-Related DNA Methylation Changes
It Is Time for a Human Epigenome Project -- Rauscher 65 (24): 11229 -- Cancer Research
Leukemia - Abstract of article: Age-related differences in leukemia biology and prognosis: the paradigm of MLL-AF4-positive acute lymphoblastic leukemia
Mitochondria, oxidants, and aging. [Cell. 2005] - PubMed Result
Modern Pathology - Abstract of article: DNA methylation in breast and colorectal cancers
NOVA | Ghost in Your Genes | Epigenetic Therapy | PBS
NOVA: Epigenetic Therapy
http://www.bio.sci.osaka-u.ac.jp/bio_web/lab_page/ogura/photo-neuron.jpg
PLoS Biology - DNA Methylation Profiling of the Human Major Histocompatibility Complex: A Pilot Study for the Human Epigenome Project
Program | The First Annual Iberian Congress on Anti-Aging Medicine and Biomedical Technologies
Proteome analysis of human hair shaft: from protei...[Mol Cell Proteomics. 2006] - PubMed Result
Reversal of Maternal Programming of Stress Responses ... Altering Epigenetic Marking Later in Life -- Weaver et al. 25 (47): 11045 -- Journal of Neuroscience
Title: Epigenetics and aging: the targets and the marks. Source: Trends in genetics [0168-9525] Fraga yr:2007 vol:23 iss:8 pg:413 -8
The DNA methyltransferases of mammals. [Hum Mol Genet. 2000] - PubMed Result
The emerging role of epigenetics in cellular and o...[Exp Gerontol. 2003 Nov-Dec] - PubMed Result
Transposable Elements: Targets for Early Nutritional Effects on Epigenetic Gene Regulation -- Waterland and Jirtle 23 (15): 5293 -- Molecular and Cellular Biology
Transposable Elements: Targets for Early Nutritional Effects on Epigenetic Gene Regulation -- Waterland and Jirtle 23 (15): 5293 -- Molecular and Cellular Biology
NCI Epigenome Project