Sports and Genes (2004, Geneva)


Sports and GenesAISTS 2004-09-22

Bengt Kayser

Rankinin, Perusse, Rauramaa, Rivera, Wolfarth, Bouchard – Human gene map for performance and health related fitness phenotypes: the 2003 update Med, Sci. Sport, Ex, vol 36 9 1451-69,

Altitude tents, induce endogenous production of red blood cells

Marathon mice engineered for extra endurance CNN Aug 23

Schuelke et al. Powerful Genes – myostatin rgulation of human muscle mass, Journal p..2682-88 Mcnally, e.m. -    four year old child with unusually large muscle mass

should we exclude this person from comp?

Geneticists engineer marathon mice Helen pearson news@nature.com 2 strains – one dies, the other runs faster for longer

Bob Goldman, 1984, Death in the Locker room

Lane, T. www.theage.com A future of jocks, genes and jingoism Peter fricker – Athens Olympic Team -    identifying athletic genes

Iannis Pitsalidis International Centre for East African Running Science

Why do Kenyans and Ethiopians Win all the time?

Why do Kenyans and Ethiopians win Most of the Time!

Last time an African won marathon was in 1968

Though performances are remarkable

Can genetics explain t dominance of east Africans in world athletics?

Scott,  … and Pitsiladis Med, and Science in Sports and Ex 35 (102) 1727-1732, 2003

Heile Gebraselasie I’ve been running since I was 4 or 5, for us life was a kind of sport

Methodology

Environmental Analysis -    place of birth -    lang -    distance oand method of travel to school

Genetic analysis: mtDNA -    mitochondria are major energy producers of t body -    mitochond fn imp in ex

MtDNA useful in popn genetics -    maternal inheritance, no recombination -    ….

Wildman, DE, Uddin, M., Liu, G, Grossman, Goodman Prot Natl Acad SCi 100 12  7181-8, 2003 -    genetic difc between humans and chimps

Demographic expansion analysis

Peter Forster (2004) Ice ages, and mtDNA chronology of human dispersals Phil Trans R. Soc. Lond. B -    origins of mitochorndria and movement of genes around world

Human mtDNA tree -    many branches of tree, various mitochondrial types and we want to see whether African athletes are part of one branch specifically

MtDNA qs If mt DNA polymorp im in enduance performance..

Contral mtDNA tree -    all different types in Ethiopia – tf. Heterogenous popn -    but did not find that – no common mtDNA -    remarkable distribution -    some amazing athletes who  have genes common among Europeans, than Africa

Conclusions (mtDNA studies) -    Ethiopian athletes distinct enviro relative to ethipian -    Deep commn maternal ancestry

Human karyotype – chromosomes

Why Y? -    patrilineal inheritance -    never 2 ys in 1 cell o    fnal changes immediately subject to selection o    no recombination

Conclusions -    like mtDNA y haplotypes spread throughout tree -    ethiopian y haplotypes show assoc with elite athlete status -    how can t y be having such an effect? -    Direct effect of a genen on t y chromosome? -    Unknown subgroup of popn?

Can genetics explain dominance? -    no genetic evidence found to date! o    So far have not looked too far

They have to run back to school, otherwise they would be caned -    mm

Alex Mauron Sports, Genes, Brains and Ethics

Thomas Murray

Just deserts (1) Purely scholastic process not sport – gambling not sport Largely predictable process not a sport – treadmill test

Nothing more basic to idea of sport than notion that successs or failure ‘sufficiently’ reflects inherent merit of indiv athletes or teams

Ethics of spectator sports -    must be fun to watch in market economy -    needs to be ‘sufficiently’ fair to satisfy yearning for justice

conventional ethics of sports -    let best win -    talent, effort luck

what it means for doping -    chemistry or genetics moral shortcut -    doping disturbs ‘level playing field’

doping immemorial (iliad, chapt 23)

sport has always attracted cheating

‘Citius Altius Fortius…Purius?’ NY Acad Sci, Mgazine, aug, sept 2004-09-22- race between doping and testing

Dilemmas in doping ethics -    prevention best with practical and conceptual difficulties

doping vs honest medical treatment

increasingly doping -    uses substance naturally present in human body -    involves substances also need in bona fide medical treatment -    involves methods that are synergistic with, not alternative o, intensive training

one of major intuitions against dopingis that we substitute something easy – doping – with something that is difficult – training -    may not be true now -    requires sophisticated knowledge

doping vs honest medical treatment not possible

doping vs privacy -    doping prevention requires all-round surveillance of athletes and invasion of privacy -    incly resembles crime control, except in sport you are guilty until proven innocent

The Red Queen -    in Alice’s wonderland you have to run fast just to stay where you are, because landscape moves with you. In fact, t evolutionary arms race between predator and prey is called t Red Queen phenomenon by biologists o    used as metaphor for c-evolution of predator and prey -    sport is full of Red Queen phenomena

running 100m I less than 3 sconds -    thre is arguably an asymptotic levelling-pff of certain sports achievements, as basic human limits are increasingly tested -    puts prob of enhancement in radical light -    will  enhancement of natural hman performance become necessary to remain interesting Red Queen race between dopers and testers -    increasingly costly -    National Centre of Addiction and Substance Abuse (CASA) at Columbia Uni calls for research effort for doping prevention costing hundreds of millions of dollars -    Unforeseen ethical dilemma: would they divert funds from health-related research? If so, is sport so imp?

Mans sana (?) in corpore not so sano perhaps -    sports turned into another health problem, when sport was supposed to be health promoting

health vs hyperhealthTM -    doping prevention relies on conventional distinction between o    medical treatment – restoration of normal species-typical functioning o    enhancement – augment bodily functions (beyond normal)

distinc between therapy and enhancement in bioethics -    asked in rel to gene therapy

gene therapy – initial debate 1980s -    somatic : ok -    germline: not ok -    therapeutic: ok -    enhancement: not OK

1990s -    gene therapy failures -    normalisation of somatic gene therapy – similar to other innovative chemotherapy (pradigm of AND medicament – axel kahn) -    but normal doesn’t mean harmless (see Gelsinger)

gene doping and gene therapy -    debate concludes that somatic gene therapy on same footing as pharma therapy -    same apply to gene doping v pharma doping -    tf gene doping objectionable on same grounds

treatment ok, enhancement not ok -    no need for genetic exceptionalism

gene doping to neurodoping -    other form of doping, such as neurocognitive could becme practical before gene doping

mens ampilificata in coropre amblificato -    one of promising avenues for sports o    eg modafinil – anti-sleeping pill o    for sports involving complex tactical tasks with motor performance (tennis), neurocog enhancement (of woring memory, executive functions, motivation) by pharma means may become incly appealing neurophilosophy

3 years ago – san Francisco – neuroethics

another basic problem -    common unreflected intuition that natural = good, artificial  = bad o    fuels some of the disapproval -    does not help – sports science is highly technicised

sociologists in france – Andrieu, Rauch) -    ritual nture of intensive training -  less to do with health

another problem -    success in sports depends on talent -    first order capacitites (musculation potential, bone structure, lung capacity, etc) -    second order capacities (somatopsychic characrs such as relative intensivity to pain, endurance and the like) -    such capacities are undeserved and unequally distributed, almost by defn -    level playing field problematic

skating on thin ice- thick conceptual distinc be treatment and enhacement probc, emphasis on fairness as THE ethical motivn for doping prevention become less persuasive -    equal doping for all not appealing proposition

real reason for doping prevention less to do with fairness -    has to with health (more of a threat than intensive sports training)

Enjoy the freaks -    mark Lawson, guardian, june 7, 2003

Non-cynical -    need a more secure basis for ethics of sport -    ethical basis of doping position, relies on ethical framework for sport

Q&As

Redefine health and disease How notion of performance has evolved through time?

Whose responsibility is sport ethics?

Dietary supplements

Mattias Kamber Instit of Sport Science, Federal office of sports, Magglingen switzerland Anti-(gene)doping: The Swiss and WADA’s approach He is member of ethics and education working committee

Voices of gene doping

Devel of doping -    1807: arsenic, strychnine, opiate, alcohol, cocaine, cannabis, hypnosis -    1936: stimulants (amphetamine, ephedrine) -    1958: anabolic steroids (dianabol, testosterone) -    1968: diuretics (chlorthaldone, furosemide) -    1976:betablockers (atenolol, propranolol) -    1980: peptide hormones (HCG, hGH, EPO, insulin) -    20XX: gene doping?

Why be concerned? -    misuse of therapeutic medicine

are we ready? -    current anti-doping tools based on science applied to urine o    non-invasive o    concentration of substances (but limited info) -    anti-doping labs started biochemical methods (EPO and HGHG) and new matrix (blood)

not yet ready for genomic or proteomic analyses

challenges -    organisations and governments -    detection and research -    athletes and support personal -    pharma companies and ‘back yard labs’ -    defn and refulations

same topics as ever but more challenges

governments -    misuse of undemocratic states – need instruments – we have this from other aspects of politics – e.g. arms control,

ME: if I am an athlete who uses doping and tries to get around the rules, is that wrong? Sport relies on this

Detection and reearch -    took more than 10 yrs to develop a test for EPO from urine -    break through for hGH and blood doping in short time (blood samples)

more research (and grants) are needed -    what type of biological samples?

Athlete and support personnel -    Victor Comte, BALCO -    THG

We do not know if it is a safe drug or not?

ME: but no form of clinical trial will tell us, since ‘enhancements’ are not studied

Pharma Companies -    detection of ARANESP at OG in Salt Lake withhelp of AMGEN -    new drugs are in devel and clinical studies e.g. RSR13, CERA, DYNEPO -    pharma compnies not forthcoming in assisting sports world

early cooperation with pharma companies is essential

Laws and regulations

Gene doping has been on list since 2003. International conventions Defn of gene doping and medical application will be challenging

Pillar principle

Controls, education and information, research Doping statut of swiss Olympic Law promoting gym and sport

The government fights against doping It promotes doping prevention Formulates a doping list Trafficking, distributing, prescribing…of medication and methods for use are forbidden -    but not easy to apply -    if trafficker claims they are not elite athlete, then not legally compromised Government supports Swiss Olympic financially to carry controls Minimal standards for controls are formulated

Swiss Workshop 2002 -    create an observatory for scientific research results and its possible application for doping -    national level, we are aware of it

WADA -    now doping is most prominent threat -    no structured independent anti-doping organisation until recently -    Council of Europe was not sufficient – not international -    Banbury conference o    Conclusions and recommendations •    Gene transfer technology is beginning to show results •    Potential for misuse in sport •    Collective efforts required •    Compliance with international standards involving human subjects that prevent unethical research is essential •    Broad public discussion and devel of social and policy frameworks before abuse occur, not after •    After Tour de France, survey whether should ban it? 34% suggest we should liberalise it under medical supervision o    Swiss Assoc of Medical Doctors developed code of conduct, now for 1year, doctors have regulations for treating athletes

WADA sport specific conclusions -    if therapy, then ok

WADA calls on govs to consider: -    req detailed record-keeping in respect of all applications of gene transfer technology with independent audit -    expand standards of medical and professional behav to prohibit improper use of gene transfer -    extend civil criminal limitation periods in respect

Legislation -    close rel with govs and international orgs (Unesco, WADA) to adapt internatona and national laws to prohibit gene doping

CCES\ -    if magic pill, what would be point of sport? -    But it is not a magic pill -

ME: what should be role of ethics committee, what political importance does it have in WADA – how do others perceive its role, what is it doing?

Cost of urine test – out of comp: 200CHF, 300CF in comp – depending on substance

Sandro Rusconi Gene Doping: Not yet possible?

Now, gene doping can be used to improve performance

What is a gene? -    one gene = many functions

what is therapeutic ‘gene trasnfer’? -    transfer of functions(s), ‘somatic’ rather than germline, targeting

how far has gene therapy progressed?

Which possibilities would exist for doping with gene transfer? Conclusions and perspectives

Elite sport has become a job with dangerous side effects

Mythos ‘gene’ in good or bad sense

It has become difficult to frankly and objectively discuss about real perspectives of gene technology

Myths -    only against hereditary disease -    transmission gene alteration -    transfer must reach 100% of cells

myths gene doping -    better than conventional doping -    transmissible gene modification -    prenatal design of athletes

DNA – RNA – Protein

Genes ‘segments’ of DNA

1cm3 = 1,000,000,000 cells

the motto ‘one gene one function’ is outdated -    2-5 functions?

100,000 genes = 300000 functions

side effects of gene transferdepend on number of alternative functions of gene product -    for most cases not known

what is a gene: a regulatable nano-device for the production of RNA and proteins -    to be effective gene segment shall include o    sequences for gene regulation o    signas for manipulation/transport of RNA o    signals for translation into protein

space – regulatory – coding spacer –

reductionist paradigm of molecular biologists

gene transfer can imply- transfer of new fn or -    transfer of a compenasating, f -    or transfer of an interfering function

4 big eras of molecular medicine

1980s – genes as probes -    prenatal diagnostics -    is someone predisposed to something?

1990s – genes as factories -    biopharmaceuticals -    take segments of genome and place into cell cultures (e.g. epo)

2000s -    genes as drugs

2000 + post genomic improvements of former technologies

gene transfer – logical consequence of former progress

somatic gene therapy NFP37 somatic gene therapy www.unifr.ch/nfp37

definitions of SGT correction disoreer by somatic gene transfer -    in gene therapy genes are used directly as drugs

Pharmacological considerations difcs conventional and moleculartherapy

Classical drugs -    mw 50-500 daltons -    synthetically prepared -    rapid diffusion/action -    oral -    cellular delivery -    readily revesible

Protein drugs – from genes as factories -    mw 20000 – 100000 DA -    biologically prepared -    slower diffusion -    oral delivery not possible -    cellular delivery o    act extra cellularly -    reversible

Nucleic acids -    mw n x 1000000 Da -    biological -    slow diffusion -    slowly or not reversible

Therapies with nucleic acids (DNA) -    req special formuation -    morec complicated than conventional

Risk / benefit balance -    depends on adopted therapy and targeted disease

why somatic? -    germ line cells: hereditary -    somatic: all other cells

4 qs about gene therapy -    efficiency of gene transfer -    specificity og gene transfer -    persistence of gene transfer -    toxicity of gene transfer

variables? -    which disease/gene/vector/organ/tissue/delivery method

3 main anatomical

ex-vivo -    bone marrow (allows to reconstitute immune system)

in-vivo -    local delivery o    cancers o    e.g. brain, muscle, eye, joints, tumors -    systematic delivery o    egs intravenous, intra-arterial, intra-peritoneal

2 classes of vector -    non-viral o    transfection o    nuclear envelope barrier! -    Viral o    Infection

Why viruses so attractive? -    they know better how to transfer DNA than us

Efficiency of transfection with recombinant DNA compared to infection with recombinant viruses

Mini-list of popular gene transfer vectors -    adrenovirus -    adreno-associated v -    retrovirus -    lentivirus -    Naked DNA o    Liposomes

Recap: limitations of current transfer vectors - can contain only certain amount of DNA in virus

there is no perfect vector

costs of each path through to clinical phase III to registration $80m -    still don’t know if would be registered – only 1 in 4 is registered

trends of clinical GT experimentation -    as of june 2003, 918 cumulative protocols -    4500 treatments -    www.wiley..com/genetherapy

1990, 1993, 2000 / ada deficiency f Anderson, m blasé, c bordignon

1997, 2000 j isner, I baumgarter, 1998, 2000

1998 restenosis v dzau

2000 hemo.. m kaey Jan 2004 – first product gendicine, by Sibiono inc 2004

No god medication without side effects

Most feared side-effects -    immune response to vector -    immune response to new or foreign gene product -    gen toxicity of viral vectors -    adventitos contambinants in recomb viruses -    random intergration in genome (inserational mutagenesis = cancer risk) -    side effects of newly acquired gene products -    contamination of germ line cells

material side effects still virtual when GT was in early phase

5 bitter adverse situations, still only one certified deat

NY ma, 1995, R Ccrystal

Upenn, sept 199 j Wilson Jesse gelsinger

Paris oct 2 2002 a fischer Retro virus x-SCID

Paris jan 14 2003 a fishcer

Pittsburgh, may 2004 K high Aav treatment factor IX hemofphilia, patients develop anti-fix antibodies

Acculutatio of hypes and loaws: roller-coaster drive

Gene doping possible? -    gene therapy o    treatment not heritable principle works o    not yet generally available o    high risk for virtually all  types of diseases

still unreachable

3 levels of doping -    before comp (anabolic) -    during comp (performance enhancers) -    after comp (repair)

which gene transfer? -    ex vivo hematopietic -    invivo – example muscle – growth factor, anti-myostatin

doping with gen transfer, many concrete possibilities

Lee Sweeny, J. App Physiology, 96, 1097 ff (2004) – march publication -    transferred gene method -    igf-1 growth factor -    aav vector,, intra muscular -    rat model

Is rat easily transferable to humans?

Side effects of gene transfer?

Short term -    autoimmunity -    hyperimmunity -    toxic shock

long term -    fibrosis -    cancer, conventional side effects of admin factors, inaccessibility to future gene therapy interventions

Specially dangerous: -    improper procedure (unsuitable vector, insuff competence) -    inapprop material (not GMP (good manufacturing practice) conform) -    insufficient follow-up

objective limitations -    viral gene transfer (immune probs, lmited readmin, gen toxicity) -    n

most likely will not be effective and harmless

Detection? -    antibody detection -    r-nucleic acids detection -    anatomically difficult to detect, but leaves permanent genetic marking -    might require tissue biopsy

foreign gene traces short-lived in body fluitds foreign genes can be in biopsies abnormal gene products oft detectable -    if expressed in wrong tissue, can be seen

adv/disadv of gene

gene doping loses

Is big talk about gene doping just one of many psych bluffs to intimdate lower-tech adversies?

Conclusions -    gebe doping higher health risks -    biggest problem is not intrinsic to technology but bears as usual on human greed and over-ambition

sanro.rusconi@unifr.ch

Prf. Dr. Hidde J Haisma Gene doping is possible? University Center for Pharmacy Rijksuniversiteit Groningen www.rugnl/farmacie/tgm

gene therapy is protein therapy, but using genes -    when give epo as gene or protein, is same thing

genomics -    identification of genes and gene expression -    caterpiller and buttergly – have same genome, but expression is different -    same with human variation – 99% we are all same

genetic manipulation -    GMOs -    Modern biotechnology

Genetic Manipulation Herman (1989-2004) -    dutch transgenic bull -    transgene = lactoferrine to be secretedin milk

genetic manipulation of humans -    delivery for therapeutic purposes -    we don’t have pills yet – and probably would not have this

good news and bad news on gene therapy -    gene therapy works on some patients

FDA stops researchers human gene therapy expt, by deb nelson and rick weiss, wash post, marc 2 200, pa08

December7 1999, Nytimes Successful gene therapy on hemophilia

March 2 200 Nytimes Hint of success indicated in gene therapy

2982000

indications addressed by gene therapy clinical trials J Gene Medicine www.wiley.co.uk/genmed/clinical monogeneic diiseas 9.8 (n=90)

requirements

genetic material to treat disease -    dna, rna

method of delivery -    viral or nonviral

how to get the dna? – internet!

How to get dNa of erythropoietin Go to anymolecular boil sites and it tells you Chromosome 7, location 7q22, geneID 2056

Gene therapy vectors

Vector – adv – diasdv Naked dna – no limitation on size – low transduction efficiency, no integr Liposomes – no lim on size – low trans and eff, no integration Retro

Gene therapy vectors Non-viral

Monogenic diseases

Factor IX in haemophilia B 9 different factors to induce clotting -    one missing or too many cause problems

Kay et al nature genetics, 2000 -    intro of Factor IX into muscle -    shows that it leaves the muscle cells and goes into the blood -    muscle secretes factor IX into blood -    (if this hormone like treatment occurs, you cannot tell where we injected this factor – detection not possible)

Use in sport – increased healing after trauma -    muscle injuries -    ligament and tendon ruptures -    meniscal tears -    cartilage lesions -    bone fracture

this would not be gene doping

misuse -    alternative to protein drugs -    protein identical to human endogenous protein (not possible to detect) -    gene therapy vector present locally

gene doping -    inc hematocrit, by epo -    increase blood flow by vegf -    inc muscle strength using igf-1 -    inc mucle size by inhibition of moystatin -    decrease pain by endorphins o    ME: check his refs

Some things we cannot do -    steroids not on list, since no steroid gene o    steroids made by many genes, not that far yet -    can only make proteins

Gene Doping

gene – potential – risk controlled – risks uncontrolled

epo- ++++ - +/- - ++++ IGF-1 = ++ - - - ++++ VEGF, FGF - + - /- ++++ Growth hormone - + - - - ++++ Myostatin / follistatin - ++++ - ? - ++++ Endorphins, enkephalins - + - ? - ++++

Very easy to make DNA, but might not be safe if uncontrolled

Epo -    glycoprotein hormone that stimulates production of red blood cells -    used to treat anemia resulting from o    cancer chemo o    chronic renal failure -    boost red blood cells prior to elective surgery -    is a gene we produce ourselves -    produced in kidney – stim bone marrow – inc red blood cells -    if you put in a gene, the signalling doesn’t take place -    need to ensure signalling to regulate

ye et al, science, 283:88, 1999 -    epo

IGF-1 -    greatly improved repair fn of dystrophic muscles -    promote skeletal muscle hypertrophy in young mice -    prevents muscle loss in old mice -    synergistic effects with weight-training -    rat ran up ladder, with weights on tale -    improve muscle force by 30-40%, combined with weight training, even more

Can IGF-1 over expression enhance athletic muscle performance? -    inc even without ex -    inrease rate and extent of repair following injury -    better maintenance of muscle mass, strength and speed during ex, during aging -    inc end of skeletal muscle, speed of skeletal muscle

how can we deliver this? -    inject our muscles?

Some vectors show it is possible -    systmetic delivery AAV-6 -    Anti-dystrophin lavelled muscles ffrom mice adminisrtered 1x 10E13 vector genomes of rAAV6-CK6 -    Microdystrophin and VEGF, mice we examined at 6 weeks after treatment -    Gregorevic et al, Nature Medicine, 2004

Gene therapy – risks

Person risks -    disease (vector, transgene), mutagenesis -    offspring

Mileu risks -    people: spouse, next of kin -    environment: infectious disease

as long as we treat somatic cells, no transfer to offspring

gene therapy and doping risks -    vector (contamination, replication competent virus) -    transgene (duration, amount of gene expression, auto-immune response)

detection? -    vector o    vector constitutents – requires biopsy o    vector dna – requires biopsy

-    transgene o    protein (e.g. epo, unless natural product) – yes, if in blood o    effect  - yes, if in blood

detection by proteomics -    physiological profiling o    serial blood sampling o    assessment of protein markers o    using protein maps, can see if changes have occurred o    people are within a certain range

WADA currently do not see drugs they are not looking for – profiling could help Rather than develop new assay for each new potential drugs, profiling could highlight anomalies

Preventatve measures

Regulation (gov, IOC, WADA) Codes of conduct (pharma ind, scientists) Education (athletes, supp staff, public) Detection (assay development)

When and where? -    genes and vectors are available -    plain dna is easily produced -    illlegal drugs produced and used

where? -    human and animal sports

can make epo DNA for €10-€25 -    quality control and marketing much more