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From Blood to Tooth Fairies Dr Sue Ring ResearchFest 9 th September 2012. Sue Ring Head of Laboratories. From blood to tooth fairies ALSPAC Samples The Laboratory Blood Processing Blood Related Research Findings Tooth Research. ALSPAC Samples.

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slide1

From Blood to Tooth Fairies

Dr Sue Ring

ResearchFest 9th September 2012

slide2

Sue Ring

Head of Laboratories

slide3

From blood to tooth fairies

  • ALSPAC Samples
  • The Laboratory
  • Blood Processing
  • Blood Related Research Findings
  • Tooth Research
slide6

ALSPAC Samples

6,000 milk teeth

16,000 saliva samples

100,000 Blood tubes

8,000 Placentas

15,000 urine samples

6,000 slices umbilical cord

20,000 toe nails

25,000 hair samples

slide8

LAB TEAM

14 in total : Researchers, technicians, students, Bioinformatics and IT support

Cell lines

Sample Processing

DNA

Analysis

storage facilities
Storage Facilities

1.2 Million individual tubes

30,000 DNA samples

20,000 cell lines

slide11

Cell lines

White cells

Spin blood sample at 3500rpm to separate

DNA

RNA

cell line production

Peripheral Blood Lymphocytes (white blood cells) isolated from blood

Add Epstein Barr Virus and put in solution containing sugars, protein and growth factors.

Cell line production

37oC

for 6 to 8 weeks

“feed” every 3 to 4 days

“Transformed” cell line which will grow indefinitely and provide infinite supply of cells and DNA

slide13

PLASMA

Cellular assays

Cell lines

Biomarkers

A measurable substance in an organism whose presence is indicative of some phenomenon such as disease, infection, or environmental exposure.

White cells

Spin blood sample at 3500rpm to separate

DNA

RNA

coeliac disease bingley et al 2004 bmj 328 322 323
Coeliac Disease (Bingley et al (2004) BMJ 328;322-323)
  • < 1 in 2,500 children diagnosed
  • Subclinical disease common in adults, is it in children?
  • Measured antibody (IgA-EMA) in blood samples taken at 7yr clinic
    • 5470 children tested 1% had IgA-EMA
    • Shorter and weighed less (equivalent to 9 months growth)
    • Higher incidence of diarrhoea and overall intestinal symptoms but not vomiting, stomach pain or constipation
  • Ie – similar frequency of undetected in children and adults
  • Consequences of undiagnosed condition not yet established but indicate that if screening is ever considered should start in childhood
  • Long term follow up can be looked at in CO90’s in adulthood to see if long term effects
slide16
Long-chain Polyunsaturated Fatty Acids (LC-PUFA)Lattka et al (2012) british Journal of Nutrition -in press
  • Important in maintenance of metabolism and health
  • In pregnancy important for brain growth, visual and cognitive development
  • Obtain LC-PUFA from
    • Diet – fish, eggs
    • Synthesised by body from essential fatty acids from vegetable oils
      • Controlled by enzymes – desaturases
      • Fatty Acid desaturase (FADS) genes code for the enzymes
        • Minor variants in FADS genes alter efficiency of enzymes and decrease the amount of LC-PUFA people with the variants can make.
slide17

What happens during pregnancy?

  • Looked at mother and child genes and at levels of LC-PUFA in cord blood at birth.
    • Does foetus obtain all LC-PUFA from mother or can it synthesis itself?
      • BOTH – Some types of LC-PUFA are present at levels predicted by Mum’s genotypes and others by child’s
        • Newborn metabolism greater capacity to synthesise more highly unsaturated omega-6 fatty acids and some evidence to show DHA but not all processes appear to be active
      • Also looked at blood taken at 7yrs of age when all influence of mother’s genotype had disappeared and levels of all LC-PUFA reflect child’s genotype – ie by 7yrs child’s metabolism fully functioning.

(Steer et al 2012 Human Molecular Genetics 21;1504-1512

slide18

Work to date has measured a small number of biomarkers in samples.

  • Samples from early time points are especially valuable.
  • High throughput methodology which uses very small volumes of sample to measure many different markers is now becoming affordable.

Anticipate large increase in number of projects and publications utilising biological samples in next few years.

metabolomics professor mika ala korpela
Metabolomics – Professor Mika Ala-Korpela
  • "systematic study of the unique chemical fingerprints that specific cellular processes leave behind“
  • 216 serum metabolic markers from each sample in one experiment
slide21

Tooth Research

  • Dr Louise Humphrey - Natural History Museum
  • Professor Christopher Dean - University College London
  • Teeth represent an archive of infancy – still learning what it means
  • Project is exploring how structure and chemistry of deciduous teeth relates to health and diet in first year of life
  • Teeth form during pregnancy and in first year of life – growth process recorded in marks in enamel - The Straie of Retzius which occur every 8-10 days, similar to tree rings
  • Accentuated lines - Permanent record of growth disruptions experienced while the tooth is forming, eg caused by flu or chicken pox. We are hoping to find out more about the causes of these accentuated lines.
  • Laser ablation – chemical composition
slide22

Enamel cusp

Polishedsection

(Image courtesy of Christopher

Dean

Dentine

slide23

Growth markers in enamel

Striae of Retzius:

regular growth lines

Accentuated Line:

growth disruption

(image courtesy of Christopher Dean)

slide24

Tooth Research

  • Dr Louise Humphrey - Natural History Museum
  • Professor Christopher Dean - University College London
  • Teeth represent an archive of infancy – still learning what it means
  • Project is exploring how structure and chemistry of deciduous teeth relates to health and diet in first year of life
  • Teeth form during pregnancy and in first year of life – growth process recorded in marks in enamel - The Straie of Retzius which occur every 8-10 days, similar to tree rings
  • Accentuated lines - Permanent record of growth disruptions experienced while the tooth is forming, eg caused by flu or chicken pox. We are hoping to find out more about the causes of these accentuated lines.
  • Laser ablation – chemical composition
  • Neonatal Line – Expression may relate to the type of birth and other factors such as birth weight or gestation length.
slide25

Neonatal line in a deciduous tooth

(image courtesy of Christopher Dean)