WP6 - Telomeres

Methodology project on comparing telomere measurements across

labs

Standing issues since October 2009

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NSHDNewcastle 85+ (1)

Newcastle 85+ (2)

Lothian 1936

Lothian 1921

Tel. Length (bp)

Frequency Mean NNewcastle 85+ (87y) 3725bp 588Newcastle 85+ (85y) 3914bp 754Lothian 1921 (79y) 4089bp 497Lothian 1936 (60y) 4201bp 1071NSHD (53y) 5646bp 2660

Are we handling the same source of DNA:

NSHD 53y Whole blood

Lothian 1936 – 60y Whole blood

Lothian 1921 – 79y Whole blood

Newcastle 85+ - 85y Whole blood

Newcastle 85+ - 87y PBMC

Standing issues since October 2009

Update April 2010

Whole Blood Cells (WBC) vs Peripheral Blood Mononuclear Cells (PBMC)Granulocytes -------T –cells T-cellsB- cells B-cellsMonocytes Monocytes

WBPBMC

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Telomere Length (bp)

Whole Blood vs PBMC on the Newcastle 85+ Study (85y)

Update April 2010

Update April 2010

122501050087507000525035001750

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Telomere Length (bp)

Freq

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3851 708.8 7083914 837.2 754

Mean StDev NPBMCWhole Blood

Whole Blood vs PBMC on the Newcastle 85+ Study (85y)

Why through all the cohorts, men have consistently longer telomere than women

when the general view is that women have longer telomere than men?

David Bann (UCL), Mike Gardner (Bristol) and Laura Wiley (Newcastle)

Telomere search strategy for Medline

10149 studies based on the title and abstract

1221 studies for more detailed evaluation (full paper)

Standing issues since October 2009

Update April 2010

Standing issues since October 2009

Is the Lothian Cohort healthier than the Newcastle 85+?

122501050087507000525035001750

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Telomere length (bp)

Freq

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4089 412.6 4973914 837.2 754

Mean StDev NLothian 1921 at 79yNewcastle 1921 at 85y

4209 564.1 1463725 399.9 588

Mean StDev N

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Telomere Length (bp)

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Lothian 1921-87yNewcastle 1921-87y

Update April 2010

Standing issues since October 2009

Validation of telomere measurements by real time PCR between labs

Paul Shields (Glasgow) and Carmen Martin (Newcastle)

150 DNA samples tested in both labs (Plates 17 and 18 of NSHD Cohort)

100007500500025000

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Newcastle Normalized Telomere length (bp)

Glasgow Normalized RatiosR= 0.463, p=0.060

Update April 2010

International Inter-lab Comparison of Telomere Measurements

- Same DNA samples (10 cell lines)

- Three different techniques applied: Southern blot, Real Time PCR and STELA

- Blind measured by 9 labs (coding by Andrew Wong, London, MRC)

Dr Sophie BekaertGhent , Belgium (Southern blot)

Prof Dr Petra BoukampHeidelberg, Germany (Southern blot)

Dr Pim van der HarstGroningen, Netherlands (Real time)

Dr. Duncan BairdCardiff , UK(STELA)

Prof Nilesh SamaniLeicester, UK (Real time)

Dr Paul G ShielsGlasgow, UK (Real time)

Dr Richard CawthonSalt Lake City, USA(Real time)

Prof. Göran RoosUmeå, Sweden(Real time)

Dr Carmen Martin-RuizNewcastle, UK(Real time)

Update April 2010

Standing issues since October 2009

-Technician recruited last year went on maternity leave in October 2009 and she has just been replaced.

- Hertfordshire DNA samples (700 + 300)

- Caerphilly DNA samples (1000+1000)