Gene Doping ConferenceFlorence, Italy 2008.10.25

15:30            Welcome

Maria Luisa Giovannucci Uzielli

MODERATORS: Angela Schneider and Mauro Giacca

Arne Ljungqvist 1989 following Seoul 1988

Florence was first anti-doping conference in 1988

HM&R committee -    list: 11 -    lab: 8 -    tue: 6 -    gene: 5

Gene Doping Panel -    Ted -    Odile Cohen-Haguenauer, France -    Lee -    Doug Wallace, California -    Kurt Zinn, Alabama

$7m annual budget (1999)

ME: Today?

Torino 2006 - Austrian skiers  - IOC had info from WADA on the team – training site found haematological lab – found no athletes – WADA sent message to IOC – Austrias already in Italy – IOC informed Italian authorities of suspicion – engineer was known to be around athletes – if IOC had not acted, would have been blamed – IOC indicated that it would make a surprise test on team in 48hrs – Italian authorities came back in 24hrs indicating coordinated action – troops were guided to find the skiers – Austrian team panicked and jumped, some escaped, some found – Italian authority were investigating – all tests were negative – but Italians found material (illegal in italy) – would not had been discoeverd had their not been a law in place – based on this, IOC could ban number of Austrian athletes and officials – Austrian Olympic Committee fined $1m to IOC – Italian law prosecuting now

China – did have similar law in place to allow Chinese to do the case –

Gene Doping -    transfer of cells or genetic elements or the use of cells, genetic elements or pharmacological elements to modulating expression of endogenous genes t having the capacity to enhance athletic performance, is prohibited

can add something to list if makes 2 out of 3, but can

for gene doping is required that is performance enhancing – exceptional case, other elements do not have this stipulation

use of medical treatment without medical indication

PPARdelta agonists PPARdelta-AMP-activated protein  kinase (AMPK) Axis aonists (eg. AICAR)

Unacceptable for medical and ethical reasons

St Petersburg 2008 Particular concern about internet distribution Boundary between therapy and enhancement

ME: what is your best guess on the schedule for how close

Applications and Grants -    27% of WADA budget goes to detection research

genomics, proteomics, microarray, imaging/detection, markers, bioinformatics

The Future -    WADA is certainly the lead agency – in fact the only one that I know of – in the application of modern molecular genetics and DNA technology to t devel of improved methods for detection in doping and in averting the use of gene therapy approaches to doping’’ (Ted, friedmann in WADA Play True, 2007).

In certain circumstances we are ahead of cheaters – no evidence of use in sport yet, but have prmade progress

Francois XXX – here at conference – testing Tour de France –

We are ahead of those who might try to do this

Ted Friedmann Gene Modification in Sport: Doping and Detection

First meeting of this topic outside of WADA’s organization

Certainly possible that gene therapy has been successful and has cured people, but few are willing to say this in public, since many cases have shown disease symptoms later on (SCID-II)

Successes SCID

Progress in treatment of childhood blindness by gene therapy (Leber’s amaurosis in children) -    little doubt that has been effective.

Gene therapy as an immature technology is reserved for serious disease – “for the moment”

Springsteen - repoxygen

Bhasker and Friedmann (2008) Insulin-like growth factor-1 coordinately induces the expression of fatty acid and cholesterol biosynthetic genes in murine C2C12 myoblasts BMC GENOMICS, in press

WADA informatics  digital data processing platform

Policy and Ethical Problems caused by anti-doping regulation

WADA Code 2009 –

17:30–18:00

Pietro Mennea,  1980 Olympic Champion (gold medal 100m), expert of Law "Doping in Sports Between National and EC Regulations"

can you win without drugs? Yes.

I never had any major injury, no muscle tear -  I was regarded as the hardest worker in the world of athletics –

So, I still believe you can win without drugs

I was introduced to Cassius Clay – and he was told that he was the fastest man in the world – Clay said ‘You’re white’ and I said ‘yes, I’m black in side, blacker than you are’ – regardless of genetics it’s possible

Even if not predestined

Comparison between Bolt and myself – win less in Europe? No. today, Afro-Americans are winning today. Jamaicans.

What have we done for anti-doping? Before WADA was in hands of sports agencies Previously handled by IOC Purpose was to control laws

Matter of public health cannot be left to sports – state must be involved.

WADA cannot succeed in battle by themselves

We have to believe in this.

2003 WADA Code in Copenhagen – convention against doping -

BALCO –  THG – Marion Jones not positive, failed to be truthful in court of law

We need a criminal law – marketing now more than it was 20 years ago, in the hands of organized crime – mafia, Italian-American, Chinese, Russian – in sale of substances

Sale in gymnasia, spas, brings in money for criminal activity

Only if the state is involve through law Community Criminal Law – now 27 nations agree – uniform the control of crime Law did not pass -    up to now, nobody has put forth this kind of request. Nobody asked for passing of criminal law.

Countries with laws 1999 Denmark (recently amended) 2000 Italy 2006 France 2006 Spain – effective 2007 – spain one of the last due to Operacion Puerto 2007 Austria

doping will never be defeated because BALCO  for eg – there will always be people like that – problem of making money

people in sport surrounded by businesses

effects

18:00-18:15

Federico Bussolin, 1st at Eurojunior Championship 2008 (200m Butterfly) 2nd at World Championship 2008 (200m Butterfly) "Testimonies by a Young Athlet"

1999 WADA established I was an MEP and witnessed creation of WADA, which is doing an enormous amount of work against doping.

18:15-18.30 Voula Kozompoli, silver medal at Athens 2004 Olympic Game (Women Waterpolo), Captain of Olympic Greek Waterpolo Team at Beijing Olympic Game "Testimonies by an Olympic Athlet"

18:30-19:15

Andy Miah “Genetic Enhancement via Genetic Selection: Bioethical and Biolegal Boundaries”

19:15-19:40  “This is Florence” A short video-presentation

19:40-20:30  Buffet

20:30           Guided visit to the Palazzo Vecchio

Sunday 26 October 2008, Florence Convention Center, Piazza Adua 1

7.30               Registration  – Poster Exhibition Open

MODERATORS: Theodore Friedmann and Philippe Moullier

8.30 - 9.00      H. Lee Sweeney “Gene doping: How could it be done and when  might it happen?”

9.05 - 9:35

Hidde Haisma “Gene Doping- Fact or Fiction”

any students in biology can do this.

9.40 – 10.10   Judith Hall “Surprises and Secrets of the Human Genome- Things to keep in Mind When Looking for Gene Doping”

In the long run, gene therapy will be untraceable

Systems biology -    control of gene expreion

2003 Human Genome project completed, despite 2001 publication

20,000 genes (200,000 proteins)

only 4% of human genome are genes -

protein expression is key

15% of Asians who metabolise testosterone differently

huge variation within normal functioning

10 years from now, could have your genome sequenced for $1,000

pharmacogenomics – drug response - 5% of people have no response - 5% over respond - Must be related to metabolizing pathway

Ethical requirements for human drug research

Athleticogenomics – what factors are important – what research is needed

Genes, Metabolism and systems of interest Muscle, vaculature, nerves, bain, lung, growth factors, repair mechanisms, sources of energy (mitochondrial variation) -    in some places they uncouple ther mitochondrial to make heat, but an athlete would not want to do this

Systems Biology -    affecting one area affects another -    nothing is in isolation

possible to enhance one pathway

we are a long way from understanding secondary effects

in development of life, you use different genes – eg. embryo uses different set of genes from adolescent, etc -    haemoglobin different between foetus and adult -    going back to foetal pathways could be important

gene control -    expressing genes

Chromatin Structure – DNA

Micro RNA – control a whole set of proteins – not traceable

Other considerations -    diet, transgenerational effects

Diet -    north American diet deficient in folic acid -    don’t know enough about diet Agouti mice -    mutation, gene involved is an imprinted gene (only maternal inheritance), if you give mum a lot of folic acid, then can benefit the offspring -    folic acid metolates intruder by turning it off

Gl flora – by-products shift metabolism and ould be used to enhance performance

What gets inherited is not a deterministic genotype,but rather a genotype that encode a potential range of phenotypes (Gilbert 2000)

Drug effects or gene therapy may be passed on to the next generation

Your grandmother

Grandmother – mother – child -    the genes of ‘child’ were being developed in the mother’s womb, during gestation in the grandmother

chimera

mosaicism -    arises brand new Microchimerism -    find babycells in mother’s blood -    can find baby cells in mother blood, but also cells from every pregnancy. -    The cells stay there for a life time -    Stem cells

FetoMaternal cell trafficking (Bianchi XXX)

Fetal Maternal Microchimerism - cells can play a role in repair / Future: Easy to identify people with genetic potential for sports, Need huge amount of research to examine effects / Likely to do DNA profiles / Likely to need more tisues / Ormal human variation is enormous and mosaicism and natural microchimerismare unversal / Gene therapy today has signature but in the future my be imposible to test / Unethical to use such therapies without extensive research / Undoubtedly there will be individual variation in response

ME: scenario –

Questions & Answers

Is

In an adult, 1 in 10,000 cells is a stem-cell

Lee nelson

Diane Bianchi

One athlete claimed on ‘vanishing twin cells’

P. Mullier: As soon as we showed results, we had athletes come to the lab asking about it

Lee: real threat is from scientists who want to make money off athletes. India and China – many people who will charge, even though no benefit.

Michael Turner: vast majority of products not by elite athletes, but by people in gym – cosmetic result. Stem cell treatment widely used in horse racing in repair of tendons – we focus too much on winner of gold medal.

10.15 – 10.35  Discussion

10:40 – 11.10  Coffee break

11:15 – 11:45  Alun Williams “Human Genetic Variation and its association with physical performance phenotypes”

23 genes – 1 in 7milion chance of existence of 1 in UK -    but expect many others influence

ME: the individual who has 23 of the genes is more likely to be a good endurance athlete, but is it also likely to be more capable than an individual with 10 or 1? – is there cumulative effectiveness?

Questions & Answers

Jim: low probability of having ultimate athletic genotype, probability

Q: association studies – today, concensus that only way to do it is whole gene snip analysis – has anyone started such an approach?

11:50 – 12.20  Mauro Giacca “AAV vectors as highly effective tools for IGF-1 gene doping”

12:25-12:55    Giuseppe Lippi “Gene doping, hypoxia and enhanced erythropoiesis”

gene therapy for lng-term expression of erythropoietin in rats Proc Nat Acad Sci, USA 1995, vol92, pp 8055-68

13:00- 13:20   Anna Baoutina (scheduled scientific contribution) "Evaluetion of an approach to directly detect gene doping using EPO as a model system"

13:20-13.40    Maria Minunni (scheduled scientific contribution) "Bioanalytical approach based on affinity sensing as promising tool for gene doping detection"

13:45 – 14:30  Lunch

MODERATORS: Arne Ljungqvist, H. Lee Sweeney

14:30-15:15     Posters Presentation (eight minute each, with slides)

HFL Sport Science LC/MS/MS and quantitative proteomics Pamela Brown

Protein in serum/plasma

15.15 – 15:45   Philippe Moullier “Genetic Doping with erythropoietin cDNA in primate muscle is detectable - Part I”

15:45 – 16:15   Françoise Lasne “Is EPO Genetic Doping possible b direct approach?

16.20 – 16:45  Tea break

16:50 – 18:50   Round Table: “Gene Doping: what is possible and what is not” Theodore Friedmann, Françoise Lasne, Arne Ljungqvist, Judith Hall, Alun, Williams, H. Lee Sweeney, Hidde Haisma"

19:00 - 20:00   Buffet

20:00              Guided Visit to the National Museum of Florence, "Bargello", especially open for the Symposium Participants

Monday 27 October 2008, Florence Convention Center, Piazza Adua 1

7:30              Registration  – Poster Exhibition Open

MODERATORS: Judith Hall, Alun Williams 9:15 - 9:45     Angela Schneider “Gene Doping: Ethics and Privacy Rights”

athletes not deprived of rights if demed ineligible

must still respect human rights

David Suzuki – challenged research on racial profiling in public sphere

10.00-10.40 POSTER PRESENTATIONS Jim Rupert Indirect SAGE analysis – epo Blood based test Aim to see if we can distinguish between epo and altitude chamber Epo expression in absence of hypoxia response is the main interest.

Q: if athlete goes to altitude and uses epo, can you discover? A: Assumption is that if go to altitude, you don’t need epo

Valeria Mastellone Vincenzo

Relationship between ACTN3 and ACE I/D

ACTN3 gene directly involved

ACTN3 R577CX polymorphism conists of a converstion of an arginine residue to a premature stop condon at resdue 577

Frequencies of allelic frequences in ialian population compared to elite athletes

ACE

Gayagay (1998), Alvarezz (2000), Nazarov (2001), Scanavini (2002) -    indicates statistical significance

10:40 – 13:30  Round Table: “Genetically Modified Athletes: Bioethics,Technology, Legal Implications” Angela Schneider, Andy Miah, Pier Francesco Mannaioni, Michail Shapiro,, Giuseppe Lippi, Judith Hall, Hidde Haisma, Domenico Giampietro Pellegrini

13:30 – 14:30  Lunch

14:30 – 14:50  Presentation of Posters  (eight minutes each, with slides)

14:50            Conclusions Theodore Friedmann Arne Ljungqvist Andy Miah Giuseppe Pieraccini Giorgio Galanti Massimo Gulisano Maria Luisa Giovannucci Uzielli

15:50            Symposium closed