From Blood to Tooth Fairies
Dr Sue Ring
ResearchFest 9th September 2012
Sue Ring
Head of
Laboratories
From blood to
tooth fairies
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ALSPAC Samples
The Laboratory
Blood Processing
Blood Related Research Findings
Tooth Research
ALSPAC Samples
• Helen Pearson (2012), Nature 484, 155-158
ALSPAC Samples
6,000 milk teeth
16,000 saliva samples
100,000 Blood tubes
8,000 Placentas
6,000 slices
umbilical cord
15,000 urine samples
25,000 hair
samples
20,000
toe nails
Background to
Laboratory
LAB TEAM
14 in total : Researchers, technicians, students, Bioinformatics and IT support
Sample
Processing
DNA
Cell lines
Analysis
Storage Facilities
1.2 Million individual tubes 30,000 DNA samples
20,000 cell lines
Blood Processing
Cell lines
White
cells
DNA
RNA
Spin blood
sample at
3500rpm to
separate
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.
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
PLASMA
Cellular assays
Cell lines
Biomarkers
White
cells
DNA
RNA
Spin blood
sample at
3500rpm to
separate
A measurable
substance in an
organism whose
presence is
indicative of some
phenomenon such
as disease,
infection, or
environmental
exposure.
Blood Related
Research Findings
Coeliac Disease (Bingley et al (2004) BMJ 328;322-323)
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< 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
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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
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.
What happens during pregnancy?
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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
• 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
• "systematic study of the unique
chemical fingerprints that
specific cellular processes leave
behind“
• 216 serum metabolic markers
from each sample in one
experiment
Tooth Related
Research
Tooth Research
Dr Louise Humphrey
Professor Christopher Dean
- Natural History Museum
- 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
Enamel cusp
Polished section
Dentine
Image
courtesy of
Christopher
Dean
(
Growth markers in enamel
Striae of Retzius:
regular growth lines
Accentuated Line:
growth disruption
(image courtesy of Christopher Dean)
Tooth Research
Dr Louise Humphrey
Professor Christopher Dean
- Natural History Museum
- 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.
Neonatal line in a deciduous tooth
(image courtesy of Christopher Dean)