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WP6 - Telomeres Methodology project on comparing telomere measurements across labs

WP6 - Telomeres Methodology project on comparing telomere measurements across labs. Standing issues since October 2009. Frequency. Mean N Newcastle 85+ (87y) 3725bp 588 Newcastle 85+ (85y) 3914bp 754 Lothian 1921 (79y) 4089bp 497 Lothian 1936 (60y) 4201bp 1071

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WP6 - Telomeres Methodology project on comparing telomere measurements across labs

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  1. WP6 - Telomeres Methodology project on comparing telomere measurements across labs

  2. Standing issues since October 2009 Frequency Mean N Newcastle 85+ (87y) 3725bp 588 Newcastle 85+ (85y) 3914bp 754 Lothian 1921 (79y) 4089bp 497 Lothian 1936 (60y) 4201bp 1071 NSHD (53y) 5646bp 2660 300 250 Newcastle 85+ (2) Lothian 1921 200 150 Lothian 1936 100 50 NSHD Newcastle 85+ (1) Tel. Length (bp) 2000 4000 6000 8000 10000 12000

  3. Standing issues since October 2009 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 Whole Blood Cells (WBC) vs Peripheral Blood Mononuclear Cells (PBMC) Granulocytes ------- T –cells T-cells B- cells B-cells Monocytes Monocytes Update April 2010

  4. Update April 2010 Whole Blood vs PBMC on the Newcastle 85+ Study (85y)

  5. Whole Blood vs PBMC on the Newcastle 85+ Study (85y) 140 Mean StDev N 120 PBMC 3851 708.8 708 Whole Blood 3914 837.2 754 100 y c n 80 e u q e r 60 F 40 20 0 1750 3500 5250 7000 8750 10500 12250 Telomere Length (bp) Update April 2010

  6. Standing issues since October 2009 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) Update April 2010

  7. 140 Mean StDev N Lothian 1921 at 79y 4089 412.6 497 120 Newcastle 1921 at 85y 3914 837.2 754 100 y c n 80 e u q e r F 60 40 20 0 1750 3500 5250 7000 8750 10500 12250 Telomere length (bp) Standing issues since October 2009 Is the Lothian Cohort healthier than the Newcastle 85+?

  8. 70 60 50 40 30 20 10 0 1400 2100 2800 3500 4200 4900 5600 Update April 2010 Mean StDev N Lothian 1921-87y 4209 564.1 146 Newcastle 1921-87y 3725 399.9 588 y c n e u q e r F Telomere Length (bp)

  9. Glasgow Normalized Ratios 2.5 2.0 1.5 1.0 0.5 0.0 0 2500 5000 7500 10000 Newcastle Normalized Telomere length (bp) 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) Update April 2010 R= 0.463, p=0.060

  10. 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 Bekaert Ghent , Belgium (Southern blot) Prof Dr Petra Boukamp Heidelberg, Germany (Southern blot) Dr Pim van der Harst Groningen, Netherlands (Real time) Dr. Duncan Baird Cardiff , UK (STELA) Prof Nilesh SamaniLeicester, UK (Real time) Dr Paul  G ShielsGlasgow, UK (Real time) Dr Richard Cawthon Salt Lake City, USA (Real time) Prof. Göran Roos Umeå, Sweden (Real time) Dr Carmen Martin-Ruiz Newcastle, UK (Real time)

  11. 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)

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