Fetal programming

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Ontogenesis PhylogenesisDevelopmental Plasticity Devo-Evo

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XX Century Epidemiologic

Transition

Environment

ERNESTO BURGIOERNESTO BURGIOISDE Scientific CommitteeISDE Scientific Committee ECERI - ECERI - European Cancer and European Cancer and Environment Research InstituteEnvironment Research Institute

From Genetics to Epigenetics

Evolutionary Medicine

Is there a Project ?Is there a Project ?

La transizione epidemiologica del XX secolo: dalla genetica all'epigenetica

Insulino-resistance Diabetes

Cardiovascular Diseases

The XXth CenturyEpidemiological transition

A switch from a prevalence of acute, exogenous (infectious and parasitic) to a prevalence of chronic endogenous diseases (immunological, neurodegenerative, endocrine, cardiovascular and neoplastic..)

Barker Hypothesis (1989)

(1989)

ab

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Let’s begin by the seventhseventh key word..

This is a graph taken from a famous article published 10 years ago on NEJM, showing the rapid decrease of the infectious/acute diseasesand the simultaneous increase of the chronic/inflammatory diseases in the North of the World

This is a graph taken from a famous article published 10 years ago on NEJM, showing the rapid decrease of the infectious/acute diseasesand the simultaneous increase of the chronic/inflammatory diseases in the North of the World

This is a figure taken from the same article, showing the presence

of a South North Gradient concerning this epidemiological transition

This is a figure taken from the same article, showing the presence

of a South North Gradient concerning this epidemiological transition

ENVIRONMENTAL FACTORS >> DNA

TIPE I DIABETES

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& Non-Communicable Diseases

Here we see that environment and lifestyles have, in this epidemiological transition, a much greater role that the DNA: migrants from the South to the North will soon get sick of the typical, chronic “Non-Communicable Diseases”

Here we see that environment and lifestyles have, in this epidemiological transition, a much greater role that the DNA: migrants from the South to the North will soon get sick of the typical, chronic “Non-Communicable Diseases”

• Perchè la celiachia (glutine + DQ2-DQ8 + Permeabilità Intestinale + Gut Ecosystem/TLRs) ha un incremento di circa il 10% annuo in Italia..?

• Perché asthma/allergie di I tipo GC sono passate da una prevalenza dell'1% a 25-30% in meno di un secolo (allergens + genes + flogosis/remodeling) ecc..

• Che legame c'è tra l’incremento epidemico di obesità.. sindrome metabolica.. diabete II.. aterosclerosi e patol.cardiovascolari (genes+epigenetics/phlogosis + TLRs)

• ENVIRONMENT HEALTH• Genome and Epi-genome• The Epidemic Revolution of XXth Century• 3 PARADIGMS

Barker HypothesisHygiene HypothesisSystemic-chronic (low grade) Inflammation

• Back to the NEO-LAMARCKIAN PARADIGM ENVIRONMENT Epigenetic Changes Fetal Programming

Fluid (Epi)genome

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Obesity Trends* Among U.S. Adults 1985

Source: Mokdad A H, et al. J Am Med Assoc 1999;282:16, 2001;286:10.

In the next 6 slides (taken from JAMA) we’ll follow, in quick succession, the dramatic, TRULY EPIDEMIC SPREAD

In the next 6 slides (taken from JAMA) we’ll follow, in quick succession, the dramatic, TRULY EPIDEMIC SPREAD

Obesity Trends* Among U.S. Adults 1987

Source: Mokdad A H, et al. J Am Med Assoc 1999;282:16, 2001;286:10.

Obesity Trends* Among U.S. Adults 1993

Source: Mokdad A H, et al. J Am Med Assoc 1999;282:16, 2001;286:10.

Obesity Trends* Among U.S. Adults 1995

Source: Mokdad A H, et al. J Am Med Assoc 1999;282:16, 2001;286:10.

Obesity Trends* Among U.S. Adults 1997

Source: Mokdad A H, et al. J Am Med Assoc 1999;282:16, 2001;286:10.

Obesity Trends* Among U.S. Adults 1999

Source: Mokdad A H, et al. J Am Med Assoc 1999;282:16, 2001;286:10

Obesity Trends* Among U.S. Adults 2001

Source: Mokdad A H, et al. J Am Med Assoc 1999;282:16, 2001;286:10. Today the situation has further deteriorated: further deteriorated: 65% of Americans are overweight, 35% morbidly obeseToday the situation has further deteriorated: further deteriorated: 65% of Americans are overweight, 35% morbidly obese

Jason Luo - Department of Biomedical Engineering University of California, IrvineJason Luo - Department of Biomedical Engineering University of California, Irvine

The Childhood Obesity EpidemicBM

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US DHHS, 2001; Hedley et al., 2004; Ogden et al., 2006, 2008

Matthew W. Gillman, MD, SM

in the 70s childhood obesity virtually did not exist (it was associated with rare genetic syndromes): since then the increase has been rapid and relentless

in the 70s childhood obesity virtually did not exist (it was associated with rare genetic syndromes): since then the increase has been rapid and relentless

Yet the most dramatic increase concerns children and adolescentsYet the most dramatic increase concerns children and adolescents

… with a constant anticipation of the age of onset …… with a constant anticipation of the age of onset …

The most serious consequence of the epidemic of obesity is the association with many chronic diseases: first of all with diabetes 2 ( today affecting 180 million people)The most serious consequence of the epidemic of obesity is the association with many chronic diseases: first of all with diabetes 2 ( today affecting 180 million people)

The origin of insulin resistance is due both to an accumulation of fat in adipocytes, and to the related inflammation. But, above all, both conditions above all, both conditions seem to be the product of a poor prenatal metabolic programmingseem to be the product of a poor prenatal metabolic programming…

The origin of insulin resistance is due both to an accumulation of fat in adipocytes, and to the related inflammation. But, above all, both conditions above all, both conditions seem to be the product of a poor prenatal metabolic programmingseem to be the product of a poor prenatal metabolic programming…

The link between obesity and metabolic 2 diabetes is complex …The link between obesity and metabolic 2 diabetes is complex …

A Silent PandemicIndustrial Chemicals Are Impairing

The Brain Development of Children Worldwide

For immediate release: Tuesday, November 7, 2006

Seven years ago two well known experts in Environmental Health, a pediatrician and an epidemiologist, launched an alarm from the pages of the Lancet, saying that a silent pandemic silent pandemic of ADHD, autism and other neurodevelopmental disorders was spreading also due to the shortage of funds in this area of research shortage of funds in this area of research

A few industrial chemicals (eg, , lead, lead, methylmercury, polychlorinated biphenyls methylmercury, polychlorinated biphenyls [PCBs], arsenic[PCBs], arsenic, and , and toluenetoluene) ) are recognised causes of neurodevelopmental disorders and subclinical brain dysfunction. ……

* **

Grandjean P. Landrigan Ph

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Exposure to these chemicals during early fetal development early fetal development can cause brain injury at doses much lower can cause brain injury at doses much lower than those affecting adult brain functionsAnother 200 chemicals are known to cause clinical neurotoxic effects in adults… The toxic effects of much more chemicals in toxic effects of much more chemicals in the developing human brain are not known and they are not the developing human brain are not known and they are not regulated to protect childrenregulated to protect children

Exposure to these chemicals during early fetal development early fetal development can cause brain injury at doses much lower can cause brain injury at doses much lower than those affecting adult brain functionsAnother 200 chemicals are known to cause clinical neurotoxic effects in adults… The toxic effects of much more chemicals in toxic effects of much more chemicals in the developing human brain are not known and they are not the developing human brain are not known and they are not regulated to protect childrenregulated to protect children

Many scientists and researchers claim that Autism is

the fastest-growing developmental disorder in the world,

with the prevalence of diagnosis having increased prevalence of diagnosis having increased by 600 per cent over the last 20 yearsby 600 per cent over the last 20 years.. And fromfrom

1:1200 to 1:90 children 1:1200 to 1:90 children in US in the last 30 years

Many scientists and researchers claim that Autism is

the fastest-growing developmental disorder in the world,

with the prevalence of diagnosis having increased prevalence of diagnosis having increased by 600 per cent over the last 20 yearsby 600 per cent over the last 20 years.. And fromfrom

1:1200 to 1:90 children 1:1200 to 1:90 children in US in the last 30 years

AUTISME AUTISME (ASD :Autism Spectrum Disorders)(ASD :Autism Spectrum Disorders)

ASD is the fastest-growing developmental disorder in the world, the prevalence of diagnosis having increased by 600% over prevalence of diagnosis having increased by 600% over the last 20the last 20 yearsyearsNew diagnosed cases (incidenceincidence) in US increased from 15,580 in 1992 to 163.773 in 2003to 163.773 in 2003

The estimated prevalenceprevalence isof 8-12 cases/1000 children (2012)

Il 17% dei bambini US < 18°a. ha un Il 17% dei bambini US < 18°a. ha un disturbo dello sviluppo, per lo più a disturbo dello sviluppo, per lo più a carico del SN carico del SN

Disturbi dell’apprendimento

ADHD

Disordini dello spettro autistico

Ritardo mentale

Problemi comportamentali

Analoghe sono le cifre europee

Il cervello è un organo prezioso e vulnerabile e, poiché il suo funzionamento ottimale dipende

dalla sua integrità, anche danni limitati possono avere conseguenze serie ( Grandjean 2006)

Il cervello è un organo prezioso e vulnerabile e, poiché il suo funzionamento ottimale dipende

dalla sua integrità, anche danni limitati possono avere conseguenze serie ( Grandjean 2006)

ISDE Palermo - Maria Vittoria Di Matteo

Alzheimer_and_other_dementias_world_map_-_DALY_-_WHO2004_svg

In 1997, the prevalence in the US was 2.32 million

An equally dramatic trend show neurodegenerative diseases and in particular Alzheimer's disease

An equally dramatic trend show neurodegenerative diseases and in particular Alzheimer's disease

Increased amyloid A-deposition

Accumulation of hyperphosphorylated microtubule associated protein “tangles”

And even in this case we have many evidences of an early origin of the disease and of a progressive progressive anticipation in the age of onsetanticipation in the age of onset (LEARn) model

No one likes to talk about a CANCER PANDEMIC.. But we must not forget that today, practically all over the North of the world, one person out of two is likely to have a cancer ..

No one likes to talk about a CANCER PANDEMIC.. But we must not forget that today, practically all over the North of the world, one person out of two is likely to have a cancer ..

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the significant increase significant increase in the in the Less Developed CountriesLess Developed Countries & in young people all overin young people all overthe worldthe world demonstrates the limits of the SMT (limits of the SMT (necessary link necessary link between between agingaging & &CA)CA)

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(1) Cancer continuous increase(1) Cancer continuous increase

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Leucémie Leucémie lymphatilymphatiqueque

Leucémie lymphatiqueLeucémie lymphatiqueEncéphaleLymphômesNeuroblastoma-Retinoblastoma1

<1 Tessuti molli, rene (Wilms), gonadi

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ReinCerveau

It is generally argued that childhood cancers are a rare condition.

But it should be reminded

that CANCER is the main cause of death by disease in childhood

that there is a constant and significant increase of tumors in the world for this age group

that 1 : 5-600 children falls ill with cancer

That more than 13 000 children fall ill

with cancer each year in the U.S.

Bleyer A, O’Leary M, Barr R, Ries LA,editors. Cancer epidemiology in older adolescents and young adults 15-29 ears of age, including SEER incidence and survival: 1975-2000. NIH Pub. No.06-5767. Bethesda (MD): NationalCancer Institute; 2006. Jemal A, SiegelR, Ward E, et al. Cancer statistics,2008. CA Cancer J Clin 2008;58:71 – 6.

Alberto Tommasini, Laboratorio Immunologia Pediatrica, IRCCS Burlo Garofolo

is is the leading cause of death due the leading cause of death due to diseases among children over to diseases among children over the first year of agethe first year of age

(2) Child cancer increase(2) Child cancer increase

Incidenza di tumori (anno/100.000)Incidenza di tumori (anno/100.000)

We should always consider the epidemiological data in the medium and long termepidemiological data in the medium and long term, not to be deceived by the inevitable fluctuations. It's evident that the incidence rates have increased incidence rates have increased dramatically over the past 30 years in the US, from 130 to 170-180 new cases/year per million dramatically over the past 30 years in the US, from 130 to 170-180 new cases/year per million inhabitantsinhabitants (to demonstrate the importance of these data, it is useful to remember it is useful to remember that a very similar increase occurred in Europe in the same period)that a very similar increase occurred in Europe in the same period)

CA incidence in childhood and adolescence IN EUROPE ( 1970-1999)( 1970-1999)

mother

latency

A first draft of the report, published on the Lancet in 2004, demonstrated an annual increase of 1-1,5% for all cancers (with more marked increases in lymphomas, soft tissue sarcomas, tumours of the nervous system…) . But the most troubling was the increase - almost the double most troubling was the increase - almost the double - for all cancers - for all cancers in the very first in the very first year of life (apparently due to transplacental or even trans-generational exposure)year of life (apparently due to transplacental or even trans-generational exposure)

Steliarova-Foucher E, Stiller C, Kaatsch P, Berrino F, Coebergh JW, Lacour B, Parkin M. Geographical patterns and time trends of cancer incidence and survival among children and adolescents in Europe since the 1970s (the ACCISproject): an epidemiological study. Lancet. 2004 Dec 11-17;364(9451):2097-105

http://www-dep.iarc.fr/accis.htm

Cancers in adults predominantly arise in (epithelial) tissues chronically exposed to environmental stress and in cells and tissues continually urged to respond/react to it

Cancers in adults predominantly arise in (epithelial) tissues chronically exposed to environmental stress and in cells and tissues continually urged to respond/react to it

While almost all childhood cancers belong to three major groups:45% oncohaematologic tumors (leukemias and lymphomas) 25% brain tumors25% neoplastic degeneration of embryonal residuals

While almost all childhood cancers belong to three major groups:45% oncohaematologic tumors (leukemias and lymphomas) 25% brain tumors25% neoplastic degeneration of embryonal residuals

The increase particularly affects children in their first life year (the incidence rate increased by> 2%)

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Il n’y a que deux possibilités:1) l’exposition du fœtus à des agents physiques (X-rays), chimiques ou biologiques (virus) (transmis par transmission trans-placentaire ) qui puissent endamager directement le foetus 2) la transmission trans-generationelle d’une ou plusieurs lesions genètiques ou epi-génetiques

The real question is: are these (epi) mutations stochasticor provoked by environmentally induced stress ?!?

The real question is: are these (epi) mutations stochasticor provoked by environmentally induced stress ?!?

(epi)mutations in gametes

Transgenerational Carcinogenesis

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(4) Tomatis legacy(4) Tomatis legacy

Translocations typical of myeloid leukaemia, probably due to maternal exposure to some toxic compound, were shown to be present at birth in children who developed the disease years later (while not sufficient per se to cause the disease, they might increase the risk for leukaemia by inducing genomic instability) Tomatis L. Identification of carcinogenic agents and primary prevention of cancer. Ann N Y Acad Sci. 2006 Sep;1076:1-14

Translocation involving band 11q23 in AML may occur as a result of a deletion or trans-locations with a number of other chromosomes and is usually associatedwith M4 or M5 and a poor prognosis

.. the first unambiguous evidence for a prenatal origin of leukaemia was derived from studies in identical twins with leukaemia. A case of identical (monozygotic) infant twins with leukaemia was recorded in 1882, and, since that time, more than 70 pairs have been published albeit in variable detail ...

The concordance rate of leukaemia varies according to subtype and age. For infants with ALL, the rate is exceedingly high (> 50%), for “COMMON” child-ALL, is ~10%.

Adult leukaemia (ALL/ AML), in contrast, has a very low rate of concordance (< 1%).

Chromosomal translocations and preleukaemic clones arise at a substantially higher frequency (~100 X) before birth than the cumulative incidence or risk of disease, reflecting the requirement for complementary and secondary genetic events that occur postnatally. A consequence of the latter is a very variable and occasionally protracted postnatal latency of disease (1—15 years).

~1% of newborns had TEL-AML1 positive B lineage clones…which represents 100 times the incidence of TEL-AML1 positive ALL (~1 in 12,000).

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(5) Pro-leukemic translocations in foetuses(5) Pro-leukemic translocations in foetuses

Even if leukaemia fusion gene leukaemia fusion gene formation isformation is spontaneous, the risk of this occurring may be modified by other factors, including folate availability. There is dietary and genetic evidence that folate has an impact on the risk of infant and childhood leukaemia ..

MLL rearranged leukemias are associated with poor prognosis and very brief latency for MLL-AF4+ infant B ALL. This raises the question of how this disease can evolve so quickly,!

!!

Our study has supported the hypothesis that in utero exposure to chemicals causes MLL* infant leukemia and has generated specific hypotheses that require further testing. Exposure to dipyrone is widespread, particularly in Central and South America where it is available as an inexpensive, nonprescription drug. Mosquitocidals are similarly in general use in these same settings. Propoxur (Baygon°) is also widely used against cockroaches, fleas, and similar pests. Therefore, it is important that the associations observed in this study are reevaluated in an extended case-control study

Interphase chromosomes

Mitotic chromosome

Euchromatin

Heterochromatin

to recognize in the study of epigenetics the most appropriate and powerful tool to build up a new systemic and molecular model of genome ..

The first keyword: Epigenetics The first keyword: Epigenetics

finally understood as a dynamic and fluid network which can interact inside itself and with the outside

finally understood as a dynamic and fluid network which can interact inside itself and with the outside

DNAdoublehelix(2-nmdiameter)

Metaphase chromosome700nm

Tight helical fiber(30-nm diameter)

Nucleosome(10-nm diameter)

Histones“Beads ona string”

Supercoil(200-nm diameter)

Campbell NE et al (Eds): Biology: Concepts & Connections4th Edition, 2003

Euchromatin

Heterochromatin

Multiple levelsof packing are required to fit the DNA into the cell nucleus

Multiple levelsof packing are required to fit the DNA into the cell nucleus

.. it has become evident that the genome is a complex molecular system (network) made up not only by DNA sequence, but also by a dynamic and responsive structure of histones and an "epigenetic" cloud of molecules (methyl and acetyl groups, enzymes, transcription factors, microRNAs )..

In 1997 the well known molecular biologist R. Strohman attempted an oblique attackattack against the against the central central dogma dogma of molecular biologyof molecular biology; the deterministic, linear, uni-directional, and encapsulated path from DNA to RNA to proteins to phenotype..

We have wrongly extended the linear theory of the gene to the “realm” of the gene management... but the gene management is an entirely different process, involving interactive interactive cellular cellular processes that processes that display an display an interactive interactive complexity… complexity… which is which is epigenetic in epigenetic in naturenature1

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A)The Central Dogma

Francis Crick's statement of the central dogma, from an early draft of Crick (1958)available at http://profiles.nlm.nih.gov/SC/B/B/F/T/_/scbbft.pdf

Bromham L Biol. Lett. 2009;5:503-505©2009 by The Royal Society

http://news.sciencemag.org/sciencenow/2009/04/21-03.html

IN FACT GenesIN FACT Genes need to be told to switch need to be told to switch “off” and “on”:“off” and “on”:• Genes need to be told how much expression (protein) is required and where.• Genes need to be regulated Genes need to be regulated – this regulation is not performed by DNAregulation is not performed by DNA but by but by many other controls arranged in a many other controls arranged in a complex complex networknetwork• DNA has been called the Book of Life by the Human Genome Project scientists, but many other biologists consider DNA to be DNA to be simply a random collection of words simply a random collection of words from from which a meaningful story of life may be which a meaningful story of life may be assembled…assembled…• In In order to assemble that meaningful story, a living cell uses a second cell uses a second informational systeminformational system. (...) The key concept here is that these dynamic-these dynamic-epigenetic networks have a life of their epigenetic networks have a life of their own —they follow network-rules not own —they follow network-rules not specified by DNAspecified by DNA

From directing the fate of stem cells to determining how.. we grow, the genes in our body act the genes in our body act in complex networks..in complex networks.. the whole Genome is a Complex and highly dynamic molecular Network of interacting Genes and non-codifying sequences.. and proteins

Strohman R. , April 2001 Beyond genetic determinism

….Genes Know How to Network…BUT...

If the Central Dogma of Molecular Biology depicted one direction-flow of genetic information…

REVERSETRANSCRIPTION

TRANSCRIPTIONFACTORS

(and COFACTORS)

GRNetworks

SYSTEMS GENOMICS(BIOLOGY)

NATURAL GENETIC ENGINEERING

TRANSPOSABLE ELEMENTS

The “Fluid Genome”

EPIGENETIC CHANGES

ENVIRONMENT

…we now know that things are quite different: information flow is circular between genome and environment

Rudolf Jaenisch Whitehead Institute & Dept. of Biology, MIT, Cambridge, MARudolf Jaenisch Whitehead Institute & Dept. of Biology, MIT, Cambridge, MA

The Histone tailsare a critical determinant of chromatin structure

The tails of histones could be regarded as the sensory / sensory / receptive receptive componentcomponent of the genome

Histone Tails are subject to a variety of covalent modifications

Histone Code”hypothesis: modifications of the Histone tails act as marks read by other proteins to control the expression or replication of chromosomal regions

E.g. generally, Histone Acetylation is associated with transcriptionally active genesDeacetylation is associated with inactive genes(= gene silencing)

DNA methylation

Covalent modification of the DNA is important for gene silencing human cells. Most genes have GC rich areas of DNA in their promoter regions. These are referred to as CpG islands.Methylation of the C residues within the CpG islands leads to gene silencing

(highly unstable base)

Nuclear Receptor DNA Response Element

Histone Lysine Acetylation

Histone Deacetylases.

Histone Acetyltransferases;

Histone Methyltransferases

ATP-dependent Nucleosome Remodeling Complex

Many toxicants cause rapid alterations in gene expression by activating protein kinase signaling

cascades.

The resulting rapid, defensive alterations in

gene activity require the transmission of a signal directly to the histones

present in the chromatin of stress response genes:

within minutes of exposure

the phosphorylation of serine 10 of histone H3

and the acetylation

of lysines 9 and/or 14 take place

H3-K9 H3-S10

P

The “meeting-point” between the information coming from the environment and the information encoded in the DNA (hardware) is the epigenome (software): mimetic molecules (EDCs) and other pollutants or danger-signals induce the epigenome to change

Chromatin itself is the direct target of many toxicants * … toxicant-induced perturbations in chromatin structure may precipitate adverse effects.. Forcing genome to change

Chromatin itself is the direct target of many toxicants * … toxicant-induced perturbations in chromatin structure may precipitate adverse effects.. Forcing genome to change

TCDD

Viruses

HERVs

EMF

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SYNERGISM !!

“FLUID EPI-GENOME”

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We may represent the environment as a continuous stream of information (simple: photons: individual packages of E = M = Information) or complex (organic molecules, viruses etc) interacting with our cells [membrane /transmembrane receptors, signal transduction proteins, nuclear receptors, genome (DNA + Epigenome)] forcing them to adapt

We may represent the environment as a continuous stream of information (simple: photons: individual packages of E = M = Information) or complex (organic molecules, viruses etc) interacting with our cells [membrane /transmembrane receptors, signal transduction proteins, nuclear receptors, genome (DNA + Epigenome)] forcing them to adapt

The second keyword: Environment

Everyday levels Everyday levels mattermatter

At truly low levels At truly low levels ……it interferes with gene it interferes with gene activationactivation

At high levels… arsenic kills people

At moderately low levels… it causes a range of diseases

KaltreiderKaltreider et alet al. 2002. 2002

Many of these substances (Dioxins, Heavy Metals, Polycyclic aromatic Hydrocarbons) are dangerous for humans health at very low-every day-doses (which are very difficult to be assessed by the ordinary toxicological studies)

Many of these substances (Dioxins, Heavy Metals, Polycyclic aromatic Hydrocarbons) are dangerous for humans health at very low-every day-doses (which are very difficult to be assessed by the ordinary toxicological studies)

In-vitro, animal, and human investigations have identified several classes of environmental chemicals that modify epigenetic marks.. including - metals (cadmium, arsenic, nickel, chromium, CH3-mercury), - peroxisome proliferators (trichloroethylene, dichloroacetic acid…), - Air Pollutants (PM 0,1/2,5/10, black carbon, benzene), - EDCs - Endocrine-Disrupting/reproductive toxicants (DES, bisphenol A, persistent organic pollutants, dioxin).

Because these epigenetic changes are small, epigenetic changes are small, potentially cumulativepotentially cumulative, , and they may develop and they may develop over timeover time, it may be difficult to establish the difficult to establish the cause-effect cause-effect relationships among environmental relationships among environmental factors, factors, epigenetic changesepigenetic changes, and diseases, and diseases..

Conclusions: We found decreased repeated-element methylation after exposure to traffic particles.

LINE-1 methylation decreased in relation to higher black carbon and PM2.5 ambient level ,with significant associations for all the time windows evaluated and stronger effects at the

longer time windows (2–7 d).

Alu methylation showed no significant association with pollutant levels at any of the time windows evaluated

Hypomethylation of repetitive DNA sequences is expected to lead to the transcriptional activation of those repetitive sequences that still contain active promoters, potentially resulting in disruption of transcription factor balance, sense or antisense transcriptional interference, and production of transcripts

complementary to endogenous transcripts or to alterations in genomic organization and stability

A small fraction of the chromosomal DNA codes for proteins

Transposable elements can be seen as a natural genetic engineering system capable of acting not just on one location at a time but on the genome as a whole ..This dynamic view of the genome has been illustrated most impressively by Shapiro who stated that the genome is composed of modular units arranged in a “Lego-like” manner that can be altered under certain circumstances

XXI century: a dramatic transformation of the environment and uterine microenvironment

The main problem that the myriad sources of pollution across the planet is likely to causeto our health and to future generations is indeed the load of toxic molecules many of whichare incidental products of incomplete combustion.. accumulated in our bodies: EPA(Environmental Protection Agency); US Centers for Disease Control and Prevention; EnvironmentalWorking Group (EWG) (an environmental organization specialized in environmental research) are systematically biomonitoring the body burden of toxic chemical compounds, elements, or their metabolites (Chemical Body Burden), in biological substances since many years

The gift our mothers never wanted to give us

http://www.ewg.org/reports/generations/

CHEMICAL FALL OUT

ULTRAFINE PARTICLESHEAVY METALS

ENDOCRINE DISRUPTORSdioxin-like moleculles

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That’s why at present many studies in various parts of the world are evaluating the chemical body burden .. especially in women, children, embryos / fetuses, providing dramatic results.

Heavy metals, dioxins and other carcinogens released into ecosphere, and conveyed in living organisms, may bio-accumulate in tissues (bones and fat) and bio-magnify in food chains

And from tissues where they accumulate (sometimes for decades), their release is generally slow and continuous

Biomagnificazione

..these pollutants are present in blood and tissues of all men and women living in urban and industrial environments and even in the cord blood and placental and fetal tissues in more and more significant amounts year after year

What is the Global Chemical BurdenGlobal Chemical Burden..

Industrial chemicals in mothers and daughters: the pollution we share and inherit

.. metals, dioxins and other lipophilic pollutants, accumulated in maternal tissue, may pass, even many years after their absorption, into the blood and reach the fetus

Scientific studies show that fetal exposure to 800-1900 MHz used in cell phones may lead to behavioral and neurophysiological changes that persist in the behavioral and neurophysiological changes that persist in the adultadult

• Mice exposed during pregnancy Mice exposed during pregnancy have have memory impairment ,memory impairment , anxiety disordersanxiety disorders and are and are hyperactive hyperactive

• suggesting that in utero exposure to RF is a potential cause of neurobehavioral disorders.We also demonstrate an increase increase in glutamatergic synaptic in glutamatergic synaptic transmission on pyramidal cells transmission on pyramidal cells in the prefrontal cortex in the prefrontal cortex associated with these behavioral associated with these behavioral changeschanges

• suggesting a mechanism suggesting a mechanism by which exposure to cell phone radiation in utero may lead to increased prevalence of neurobehavioral disorders

Brain plasticity Brain plasticity and modulation of its structure and its functions

Motility of neurons and in particular the formation of new formation of new connections (synapses) can be connections (synapses) can be modified (perturbed) by modified (perturbed) by exposure exposure to environmental stressors

Wingate Imagining the brain cell: the neuron in visual culture. Nature Rev Neuroscience 2006; 7: 745-752.

The IndividualIndividual wiring

Early critical periods in the development of SYNAPTOGENESIS and brain functions

Formation of new synapses following stimulation..

The IndividualIndividual wiring

this is not a generic concept, not a generic concept, concerning the way in which concerning the way in which the "genetic program" the "genetic program" contained in DNA is contained in DNA is translatedtranslated, during the nine months of the ontogenetic process, in a specific complex phenotype.

on the contrary,this is a precise precise technical term that refers to technical term that refers to the ability, and at the same the ability, and at the same time to the necessity, of time to the necessity, of embryo-fetal cells to define embryo-fetal cells to define their epigenetic setting in their epigenetic setting in adaptive (and predictive) adaptive (and predictive) responseresponse to the informationto the information coming from the mother and, coming from the mother and, through her, from the outer through her, from the outer world.world.

Dioxin and Dioxin-like molecules

(Ultra)-fine particles

Heavy Metals

Polycyclic Aromatic Hydrocarbons (PAH)

Benzene

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The third key word is fetal programming

MATERNAL MATERNAL STRESSSTRESS

ONTOGENYONTOGENY

Fetal Programming

Differentiation

epi-mutations

Cellular Differentiation: an Epigentic process The actual genetic program of a actual genetic program of a particularparticular individual is actually the individual is actually the product of nine months of product of nine months of epigenetic adaptive-predictiveepigenetic adaptive-predictive “formattingformatting” of billions of cells)..

This is the stage of life which is far more sensitive to information coming from the environment information coming from the environment (particularly to maternal-fetal stress,(particularly to maternal-fetal stress, to nutritional errors, to pollutants ..)

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Nature 447, 425-432 (24 May 2007)

PLASTICITY

The fourth keyword is developmental plasticity

This image clearly shows the "power" of the epigenome and the predominant role of environmental information in the phenotypic shaping of cells, tissues , organisms .. the huge phenotypic (morpho- functional) difference between a lymphocyte and a neuron is not due to DNA, which is virtually identical in the two cells , but to the manner in which the same genome has been utilized by the two cells, on the basis of the information (positional and environmental) received formation (positional and environmental) received during the first months of life (for neuron in the first 2 years) during the first months of life (for neuron in the first 2 years) and processed by the epigenetic networks

The larvae that develop into workers and queens are genetically identicalgenetically identical. But as a result of the royal jelly diet, as a result of the royal jelly diet, the queen will develop functional the queen will develop functional ovaries and a larger abdomenovaries and a larger abdomen for egg laying

A Bee's Royal Diet

Queen Bee Larvae: Queens are raised in specially-constructed cells called "queen cups," which are filled with royal jelly.

Kucharski R., Maleszka J., Foret S., Maleszka R. Nutritional Control of Reproductive Status in Honeybees via DNA Methylation Science (2008) 319: 1827-1830

… although twins are epigenetically indistinguishable during the early years of life, … older monozygous older monozygous twins exhibited twins exhibited remarkable remarkable differences in differences in their overall content overall content and genomic distribution of 5-methylcytosine DNA and histone acetylation, affecting their gene-expression portrait.

3-year-old twins 50-year-old twins3-year-old twins 50-year-old twins

Epigenetic differences in homozygotic twins

Fraga et al., PNAS. Jul 26 (2005);102(30):10604-9..

Epigenetic differences arise during the Epigenetic differences arise during the lifetimelifetime of monozygotic twins

The chimpanzee DNA is for 98.77% identical to the human . On average, a gene encoding a protein in a man differs from its chimpanzee ortholog by only two aa substitutions .. almost one third of human genes has exactly the same protein translation as their orthologs

in chimpanzee

Species phylogeny

Orangutan Gorilla Chimpanzee Human

From the Tree of the Life Website,University of Arizona

Sanger Institute

We are quite stable (for millions of years) both

genetically and phenotypically

Evo

The fifth key word is phylogeny

And of 9 months of an 9 months of an individual developmentindividual development

of 4 billion years of molecular of 4 billion years of molecular coevolution coevolution * (in particular, our DNADNA is the product of this long journey) ..

PhylogenesisPhylogenesis

We should never forget that we are we are at the same time at the same time the productthe product

OntogenyRecapitulates(anticipates) Phylogeny

Devo-Evo

our epigenome being the product of nine months of cellular and tissue programming (adaptive to an environment that is rapidly changing)..

A major risk: the EDCs and other xenobiotics (not being the product of molecular coevolution) can interfere at this level, acting as pseudo-morphogens

OntogenesisOntogenesis

MismatchMismatch

All this means that the main variations in our phenotype (both physiological and pathological) have their origin in fetal programming and are induced by the altogether changing environment and modulated by the epigenome …

All this means that the main variations in our phenotype (both physiological and pathological) have their origin in fetal programming and are induced by the altogether changing environment and modulated by the epigenome …

Dutch famine versus Leningrad Siege

The association between poor intrauterine growth and increased risk ofdisease in later life may reflect a mismatch between the environment‘foreseen’ by the embryo/foetus, based on signals from the mother duringgestation, and the actual environment experienced in later life.

The sixth key word is mismatchThe sixth key word is mismatch

That’s why overweight, obesity, overweight, obesity, type 2 diabetes and other chronic type 2 diabetes and other chronic diseases diseases are advancing even more rapidly in the III Worldmore rapidly in the III World …owing to the very quick environmental changes in developing developing countriescountries

The sixth key word is mismatchThe sixth key word is mismatch

The problem is how to evaluate the consequences of these epigenetic genomic changes on populations chronically (daily) exposed to heavy metals and other epigeno-toxic pollutants ??!!The problem is how to evaluate the consequences of these epigenetic genomic changes on populations chronically (daily) exposed to heavy metals and other epigeno-toxic pollutants ??!!

We also know that if we expose primates (and other mammals) in the very first stages of their development to some xeno-biotics (endocrine disruptors etc) or to heavy metals (mercury, lead, cadmium) we induce fetal programming (modifying the epigenetic setting of their endocrine and immunological tissues) conditioning the whole life of their cells and tissues and opening the way to many diseases (breast cancer, Alzheimer disease..)

Agouti mice

Questi 2 topini sono geneticamente identici, ma fenotipicamente assai diversi

Quello di sinistra non è soltanto grasso e giallo (agouti), ma predisposto a diabete 2 e cancro

Tutto questo essenzialmente a causa del fatto che un singolo gene mutato può essere espresso o meno..

a seconda del fatto che sia stato o meno epigeneticamente epigeneticamente marcato /silenziato marcato /silenziato nelle nelle primissime fasi della vita primissime fasi della vita

(il topino di dx ha il gene (il topino di dx ha il gene metilatometilato grazie alla grazie alla somministrazione alla madre somministrazione alla madre

nel corso della gravidanza nel corso della gravidanza

di di genisteinagenisteina, , un componente un componente della soia che dona gruppi della soia che dona gruppi metilici) metilici)

Dr. Dana Dolinoy of Duke University

We used the Agouti mice to study how maternal nutrients and environmental factors affect the epigenome.

Specifically, we wanted to know whether a mom's exposure to a contaminant found everywhere in the environment could alter the fetal epigenome, and eventually the long-term fate of her offspring.** Bispenol A - BPA: a common chemical found in certain plastics, in many commonly used products, including food and beverage containers, baby bottles, dental sealants

About four years ago, the CDC studied approximately 400 people, and in 95

percent of these 400 people, they measured detectable levels of bisphenol-A.

THE ELUSIVE AGOUTI

In the yellow obese mouse, the Agouti gene is unmethylated and turned on all the time, while in the brown mouse, the gene is completely methylated and shut down.

There are also other mice that appear mottled in which half of the cells are methylated and shut down, and the other half are unmethylated and turned on, and these mice appear to be yellow and brown.

Transgenerational epigenetic “protection”

The extra methyl groups extra methyl groups protect the offspring from protect the offspring from obesityobesity and other adult-onset heath problems.

Surprisingly, this protection Surprisingly, this protection extends beyond the extends beyond the offspring and into the next offspring and into the next generation of mice generation of mice (the grandchildren of the initial yellow-furred mother)..

Which suggests that epigenetic modifications epigenetic modifications can be passed across can be passed across generations – a “memory” generations – a “memory” of environmental influence of environmental influence that occurred many years previously.

Environmental exposures have been found to promote several transgenerational disease states or phenotypes

The reproducibility and frequency of these disease phenotypes suggests they are likely epigenetic rather than due to DNA sequence mutations.

The exposure of a gestating mother exposes the F0 generation mother, the F1 generation embryo and the germ-line of the F2 generation.

The F3 generation would be the first unequivocal transgenerational generation not exposed.

Research has demonstrated that 90% of all male progenyfor four generations (F1–F4) developed these disease states after the direct exposure of the F0 gestating rat

This transgenerational phenotype was only transmitted through the male germ-line (sperm) and was not passed through the female germ-line (oocyte).

These animals had the following disease state frequencies; 20% tumor development,50% prostate disease, 40% kidney disease, 30% immune abnormalities, and 30% severe infertility in males from F1 to F4 generations

Obesity/Metabolic Syndrome

Neurobehavioral Deficits and Diseases

CANCER

Reproductive Diseases/Dysfunctions

Multiorgan Effects of Endocrine Disruptors

In Vitro Fertilization

Semen Abnormalities

ObesogensDOHAD

Asthma and allergies

Psychiatric Diseases

Cardiovascular Diseases

Ipertension

Developmental Time Windows of Vulnerability

Pesticides

Lung Development

Placenta: Prediction of Future Health

Materno Fetal Stress

..recently, the fetal programming mismatch theory has been transformed into the key-moodel theory of DOHAD....recently, the fetal programming mismatch theory has been transformed into the key-moodel theory of DOHAD..

OBESITYDIABESITYPANDEMICS

A new scientific truth does not triumph by convincing its

opponents and making them see the light,

but rather because its opponents but rather because its opponents eventually die, and a new eventually die, and a new

generation grows upgeneration grows up that is familiar with it.

Max Planck (1858 - 1947)