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