Hlne Blanch

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Genetics of Healthy Aging

• Hélène Blanché
• Fondation Jean Dausset - CEPH

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GEHA

• Integrated Project of EU 6thFP
• 5 years, 7.2 M , 25 Partners

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Aims of GEHA (1)

• To recruit 2650 sibpairs (10 countries, 14
  geographic areas) and 2650 matched younger
  controls
• To search for identified/putative longevity
  regions/genes and gene variants
• To test whether different populations share the
  same genes involved in aging and longevity
• To ascertain the role played in human longevity
  by chr4q, chr11p15.5 and ApoE regions

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Aims of GEHA (2)

• To identify mtDNA haplogroups and mtDNA
  mutations associated with longevity
• To identify gender-specific genes differently
  involved in healthy aging and longevity
• To stratify samples according to APOE genotype
• To develop mathematical and statistical models
  to combine genetic data with demographic
  characteristics
• To perform a longitudinal study to evaluate the
  importance of genetic factors on mortality

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Ethics

• GEHA devotes a particular attention to the
  ethical and legal issues and implications
• GEHA activity is continuously monitored by the
  Ethical and a Legal-IPR Boards

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Set up of a standardized protocol

• Informed consent, information file
• Standardized case sheet to assess and record
  which would be acceptable by local ethics
  committees
• socio-demographic information,
• lifestyle habits,
• health and morbidity,
• medication,
• sensorial function,
• cognitive function and depression,
• functional activity, physical performances.
• Training of people in charge of the recruitment

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A phenotypic database

• Centralized database in Odense and local data
  stored in each recruiting center
• EPIDATA, a standardized procedure to enter data
  online via a VPN (crypted connection) to the
  centralized database
• Restricted access by login/password

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Volunteers ID

• PID, 9 digits including 5 components 
  11 2 3333 4 5 
• 11 country
• 2 recruiting center in the country
• 3333 family ID related to the recruiting center
• 4 individual ID related to the family
• 5  check sum  character used to check the
  validity of the previous characters

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Recruitment identification of volunteers

• Identification of elderly sibpairs on census data
  or other available lists
• Contact people by phone or mail
• Visit at elderlys home, nursing home
• Identification of unrelated controls (60-75 years
  old)

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Organization of a visit

• Interview of the volunteer in presence of a proxy
  if necessary (MD or trained nurse)
• Presentation of the project
• Answer to all questions
• Informed consent is signed
• Collect of information
• Cognitive and physical tests
• Collect of biological sample
• About 2 hours per volunteer

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Biological samples and treatments

• 2 EDTA tubes labeled with bar codes (PID)
• - one is frozen without treatment
• - one is centrifuged to isolate plasma
• 3rd tube either EDTA or heparin, labeled with bar
  code (PID)
• - isolation of plasma and blood pellet
• - isolation of PBL for establishment of cell
  lines
• Cheek swab if blood donation is not possible

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Follow up of sample treatments in local labs

• At CEPH, use of Biobase I software
• - registration of biological samples at
  reception (non-identifying information)
• - registration of each step of the treatment
  process, from reception to the final derived
  sample
• - follow-up of the sample history
  identification of each single sample tube with a
  unique bar code automatically generated
• - management of stocks (storage and distribution
  of DNA samples)
• - management of biological component shipments

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KTL a centralized extraction center

• For each individual, one frozen whole blood
  sample is sent to KTL
• Extractions are performed on a Gentra automate
• Each DNA is labeled with a new ID (GID, 7
  digits). KTL has the correspondence table between
  PID and GID
• Quantification and normalization of DNAs
• Quality controls sex PCR, ID PCR
• Distribution on microtiter plates

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GEHA Genetic Platforms
Mitochondrial genome
Nuclear genome
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Genetic studies on genomic DNA

• Genome scan using SNPs (Illumina)
• Screening of candidate regions previously
  published or identified during the genome scan

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A genotypic database

• Centralized in Kiel Genedigger
• Connections between phenotypic and genotypic
  databases only allowed for partners involved in
  analyses after approval of the Steering Committee

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Merged data for analyses
Phenotypic database
Genotypic database
GID
(PID)
Genotypic WP leader
Correspondence PID/GID
Extraction lab
Sex origin genealogy (GID)
Genotypic WP leader
GID
sex
Genotypic WP leader
PID
Correspondence PID/GID
GID
Recruitment center
Recruitment center
Recruitment center
transfer of samples transfer of data
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Update on recruitment

• July 2005
• - 1538 families contacted
• 39 of contacted families gave a positive answer
• 43 of contacted families were excluded
• 7 one sib died or refused to participate
• 10 dementia
• 26 immediate refusal
• 18 of contacted families are in stand-by
• - 267 sibpairs and 267 controls are recruited
• - 731 DNA are extracted (including 14 cheek swab
  samples)
• November 2005
• - 632 sibpairs are expected.

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Update on genetic studies

• Genome scan Beginning February-March 2006 on
  800 sibpairs
• Association studies chr11p15.5 region (KRAS,
  IGF2, INS, TH), 350 markers from a 2.4 cM region
  
• Germany 377 cases and 376 controls
• Central Italy 137 cases and 168 controls
• Southern Italy 92 cases and 97 controls
• France 560 cases and 560 controls