We wish to use the ALSPAC cohort to investigate the effects of novel and previously-confirmed type 2 diabetes loci on fetal growth, growth in childhood and intermediate traits related to type 2 diabetes.

Following our analyses of the first UK type 2 diabetes genome-wide association (GWA) study, which identified 6 new type 2 diabetes genes (FTO, CDKAL1, CDKN2A/2B, HHEX, IGF2BP2, SLC30A8) [1, 2], we have performed a meta-analysis of GWA data, combining our UK study with the DGI [3] and FUSION [4] GWA scans (N=10,128 individuals and ~2.2 million SNPs, directly genotyped and imputed). We followed-up promising signals by performing replication studies of up to 63,532 independent samples [5]. Four loci showed robust evidence for association at Pless than 5x10-8. We are therefore confident that the results exceed the stringent levels of statistical support needed for genetic association studies.

The associated SNPs are as follows:

JAZF1 rs864745

CDC123/CAMK1D rs12779790

ADAMTS9 rs4607103

THADA rs7578597

The precise nature of the biological mechanism by which each of these genes predisposes to type 2 diabetes is largely unknown. We have used replication and robust statistical data, rather than biology, to identify these genes. Further analyses of their roles in intermediate traits, using ALSPAC, are likely to help to uncover further biological pathways.

In addition to the above, we would like to complete the list of currently-confirmed type 2 diabetes loci genotyped in ALSPAC by including the following:

WFS1 rs10010131

PPARG rs1801282

KCNJ11 rs5219

We therefore propose to analyse the polymorphisms in ALSPAC to test the following hypotheses:

1. Fetal genotype and maternal genotype alter fetal growth.

2. Fetal genotype and maternal genotype alter growth velocity in childhood

3. Genotype alters intermediate traits related to type 2 diabetes including fasting insulin, fasting glucose and insulin secretion (in the subset of offspring with OGTT data), triglycerides, HDL, LDL and total cholesterol, anthropometric measures including BMI, lean/fat body mass, WHR, waist circumference, skin folds where available.

4. In the event of associations with intermediate traits in mothers we will propose to test the role of maternal genotype on fetal and childhood growth and body composition and biochemistry. For example, if the SNP alters BMI the SNP can be used in a Mendelian randomisation framework to test the role of maternal BMI in fetal growth and body composition free from confounding factors such as socio-economic status.

Whether the results are negative or positive they will help in our understanding of how the novel genes function and, if positive, provide important insights into growth and other diabetes intermediate phenotypes.

* Specific ALSPAC phenotypes being considered:

To do this we would like to genotype (at Kbioscience) all ~20,000 ALSPAC samples. We will need the following phenotypes to test our hypotheses:

1. Birth weight, length and head circumference

2. Growth measures in childhood (height, weight and BMI aged 7-11)

3. Covariates of birth weight to check if genotype is acting through them: gestational age, maternal age, maternal BMI, smoking , parity, twin status to exclude non-singletons, ethnicity as genotype frequency may alter with ethnic origin and confound analyses.

4. Type 2 diabetes-related intermediate traits including fasting insulin, fasting glucose and insulin secretion (in the subset of offspring with OGTT data), triglycerides, HDL, LDL and total cholesterol, anthropometric measures including BMI, lean/fat body mass, WHR, waist circumference, skin folds where available.

In addition, we are keen to examine the KCNJ11 variant in relation to physical activity in the ALSPAC children. This is to follow up a nominal association observed in the RISC cohort (Mark Walker, personal communication).

Plans for replication:

ALSPAC will provide the largest dataset for association studies with fetal growth. However, we have access, through our own studies and extensive collaborations to samples from the Exeter family study (950 population based parent-newborn trios), Plymouth Earlybird study (300 parent-5 yr old trios - T Wilkin) the North Cumbria community genetics project (3000 mother child pairs - C Relton), the Northern Finnish 1966 Birth Cohort (4600 individuals with own birth measures) and the 1958 cohort (7000 individuals with own birth measures). We hypothesize that real genetic associations will be consistent across all these studies - i.e. even if individually studies show only nominal significance, a meta-analysis of all studies will provide highly significant results.

Aim

We aim to explore the relationship between maternal and paternal depression over time in order understand the direction of causality taking into account reciprocal effects. A further aim is to explore how this relationship alters according to child behaviour and life stresses and finally how this relationship impacts on future child emotion and behaviour.

Method

We propose analysing the ALSPAC data up to age 8 including repeated measure of maternal and paternal mood and measures of child emotional and behavioural problems. We will begin by exploring reciprocal causation between maternal and paternal depression and investigate how this varies as a function of life stress, child behaviour, marital relationship and family size. We will use multilevel models to investigate the random slopes for lags and crosslags this represents a novel method of investigating these questions with the advantage that this more flexible statistical model can model the more complex reciprocal relationships likely to arise within families.

Variables

Our main variables will be:

Maternal Mood EPDS pregnancy 18 weeks & 32 weeks antenatally 2, 8, 21 33 61 73 and 97 months postnatally

Paternal mood EPDS pregnancy 18 weeks antenatally 2, 8, 21 33 61 73 and 97 months postnatal

Child emotional and behavioural problems - Carey infant temperament 6 months and Carey toddler temperament 24 months EAS temperament 38, 57 and 69 months Rutter parent Scale for preschool chidldren 42, 57 & 69 months SDQ 47 81 97 months.

Other variables:

Age mother

Age father

Number of children in home

Primip vs multip

Nature of delivery

SES (highest educational level mother)

Quality of Marital relationship - Intimate bond measure 21 and 33 months

Life events 18 weeks 32 weeks antenatally, 2, 8, 21 33 47, 61, 73 months.

e propose to conduct a gene x gene and gene x environment analysis of candidate factors that have been shown to increase susceptibility for dyslexia and SLI in previous research or that have been chosen on the basis of their biological function. We will genotype all the available ALSPAC children DNA samples (greater than 10,500) for susceptibility variants within candidate genes and conduct the analysis using cognitive outcomes and environmental exposures already assessed in the ALSPAC project.

The genotyping will be performed using the Sequenom i-plex technology in collaboration with the Genomics Core facility at WTCHG. We already have in our laboratory the sufficient quantity of ALSPAC DNA required by this project.

We will select for analysis measures of reading, language and speech abilities, including measures of single word reading, non-word reading, phoneme awareness, orthographical skills, spelling, short-term memory (including NWR), verbal expression, comprehension and IQ. Many of these measures are the same as those used in our current dyslexia and SLI linkage and association studies. We plan to include in the analysis measures of attention and hyperactivity behaviour since attention deficit hyperactivity disorder (ADHD) shows extensive co-morbidity with both dyslexia and SLI.

The environmental factors will include measures that have already been proposed as risk factors for SLI, for example otitis media with effusion (OME) and maternal education, or associated with dyslexia, for example the home literacy environment and Omega-3 fatty acid consumption. We will consider also the child's environment, more general aspects of parenting and parent-child interaction. Statistical analysis will be based on linear regression models extended to include one or more covariates. We will also interrogate for multilocus interactions and genetic effects specific to factors underlying the phenotypes using techniques such as principal component or cluster analysis. Analysis will be performed both for single SNPs and haplotypes.

A detailed list of genotypes, phenotypes and environmental measures is given in the appendix.

Strategy details

Aim 1: To investigate whether the effect of dyslexia and SLI susceptibility variants can be modulated by different genetic factors (gene x gene interactions). We will select the maximum number of SNPs that can be accommodated in three i-plex reactions. Most of the novel dyslexia candidate genes we are proposing for this project have been selected for showing a strong signal in the recent WGA analysis we conducted in 600 individuals with dyslexia collected in Germany and in the UK as part of the collaborative NeuroDys project (www.neurodys.com). Other genes have been selected for their (i) similarity to other established dyslexia candidates (KIAA0319-like), (ii) interaction with established dyslexia candidates (we have recently identified interaction between the AP2M1 and KIAA0319 proteins) or (iii) role in neuronal migration, a biological pathway to which most of the reported dyslexia candidates seem to contribute. We will also include candidate genes for ADHD since this disorder show an extensive co-morbidity with dyslexia and SLI. SNPs have been selected either for showing significant association in previous work or for tagging the most common haplotype across the genes of interest. Statistical analysis will be conducted at different levels. We will first test for association between single SNPs and quantitative measures of reading, language and speech in a linear regression framework. This analysis will enable us to prioritise further investigations on the basis of the strength of association found with each gene. Following these initial association analyses, we plan to test for gene x gene interactions between the genotyped loci for example by integrating an epistatic component into the regression model. In this second step of the analysis we will incorporate also genes for which we have already obtained ALSPAC approval*.

A specific question we aim to answer is whether any tested SNP or haplotype can modulate the effect of the KIAA0319 risk haplotype (which we have recently identified within the ALSPAC sample) and whether it is possible to detect any specific interactions between candidate genes involved in the same biological pathways, such as between genes involved in neuronal migration.

Aim 2: To investigate the role of environmental factors in modulating the effect of genes that contribute to measures of reading, speech and language (gene x environment interactions). This analysis will aim to test whether any environmental factors can modify the effect of an individual's genetic background in either a protective or adverse manner for reading and language ability. The SNPs, haplotypes and gene x gene interactions that show significant associations in Aim 1 will be tested for gene x environment interaction using the environmental measures described in the Appendix.

Aim 3: To test whether shared genetic or environmental factors can explain co-morbidity between dyslexia and SLI and whether it is possible to identify a correlation between genetic/environment background and specific sub-groups of phenotypes. The data generated in Aim 1 and Aim 2 will be used to assess whether it is possible to identify causative factors that either have a particular effect on specific phenotypes or affect different, broader areas of cognition. Factors that are found to have a pleiotropic effect across different phenotypes will be further explored to investigate a possible role in determining the observed co-morbidity between SLI, dyslexia and related disorders such as ADHD. Since both dyslexia and SLI could be hypothesised to be caused by an underlying phonological deficit it will be interesting to see if specific genes contribute to the variation of phonological outcomes. The identification of association between single factors and multiple phenotypes will lead to the question of whether it is possible to identify additional interacting factors that result in more distinctive phenotype. The ultimate aim of this analysis is to test whether it is possible to distinguish between well-defined cognitive deficits on the basis of determined genetic/environmental background.

Oppositional defiant disorder (ODD) is the single most prevalent psychiatric illness in children and adolescent in the UK(1) and is a very strong predictor of psychiatric disorders in adulthood (2). The strongest associations of ODD are with conduct disorder, antisociality, and ADHD (1, 3-5); however, ODD is also strongly correlated with emotional disorders both cross-sectionally and longitudinally (4-6). This wide range of associations and predictions suggests that ODD plays a pivotal role in the development of psychiatric illnesses (4). Despite intense research interest in ODD, it remains unclear what the underlying causes for this breadth of associations may be. A better understanding of ODD could potentially help explain issues of particular importance to child psychiatry. Firstly, it would shed light on the puzzling rates of comorbidity in children and adolescents (5, 7), and, secondly, it could help understand developmental trajectories from early life into adulthood.

We have recently been able to demonstrate in a large national sample (the Office for National Statistics Child and Adolescent Mental Health Studies from 1999 and 2004), that ODD may comprise of three dimensions that carry relatively distinct associations with other childhood psychiatric disorders (Stringaris & Goodman, submitted). We have provisionally termed these three dimensions Irritable, Headstrong, and Hurtful, conforming to their respective predictions. The Irritable dimension, composed of items indexing temper outbursts and anger, has strong associations with emotional disorders, whilst the other two dimensions predict more strongly to conduct disorders. In addition, the Headstrong dimension appears to be most strongly associated with ADHD and the Hurtful dimension to be a strong predictor of pre-meditated aggressive offending. As a result of these findings, which preliminary evidence suggests are replicated in longitudinal analyses, we are proposing a new model for ODD. Thereby, the three dimensions we have identified represent relatively distinct contributions to ODD that we hypothesize will arise from different sets of predisposing factors.

To study these questions will require a longitudinal study with measures of ODD symptomatology in middle/late childhood, and hypothesized predictors earlier in development. ALSPAC is the only study in the United Kingdom and one of the few in the world that would allow us to address these issues.

AIMS

The aim of the present study is to identify underlying factors for oppositionality in children and adolescents. Based on our findings so far, our main hypothesis is that the three dimensions we have identified as contributing to ODD are predicated on distinct predisposing factors. We expect the distinct correlates to lie in the domains of child temperament, family mental illness, parent child relationship, and academic attainment. Our hypotheses are as follows:

Temperament: we predict the Headstrong and Hurtful dimensions to be best predicted by "difficult" temperament. In contrast, we hypothesize that the temperament underlying the Irritable dimension will be mixed, encompassing both "inhibited" and "difficult" temperaments.

Family Mental Illness: we expect that all three dimensions will be predicted by a family history of antisociality. However, we predict that a family history of emotional disorders, such as depression and anxiety, will be differentially associated with the Irritable dimension.

Family Relationships: our prediction is that family relationships are important for both the Headstrong and the Irritable dimension. In contrast, we predict that such environmental effects will be of limited influence for the Hurtful dimension. This is due to evidence we have gathered so far showing that this dimension may be linked to callous and unemotional traits; these have been shown to be mostly due to additive genetic effects.

Cognitive Attainment: similar to our predictions about other environmental effects, we expect that frustration due to low academic achievement will be most relevant for the development of the Irritable and Headstrong dimension and significantly less so for the Hurtful dimension.

ANALYTIC DESIGN

This will be a developmental follow-back study, that is, we will establish symptom dimensions at the ages of 8 and 10 years and will then seek to establish their antecedents as detailed above. The study could thus be completed entirely on the basis of existing ALSPAC data. The instrument we will use to construct the dimensions will be parent-reported ODD (awkward and troublesome) symptoms collected as part of the DAWBA assessments completed at XX and YY months (this we have also used in the ONS study).

IMPLICATIONS

We expect our findings to help better understand the development of mental illness in childhood and adolescence. In particular, we expect our results to be particularly relevant for the central question of comorbidity between mental disorders in childhood. Furthermore, if childhood oppositionality encompasses more than one dimension this could mean that differential interventions might be indicated.

Aim

The aim of this project is to investigate the associations between diet and blood pressure, measured at 5 years and 7 years in the CIF sample.

Background

Cardiovascular disease is the leading cause of death in industrialised countries, and raised blood pressure is a major risk factor(1). Geographical variations in childhood blood pressure in the UK coincide with geographical variations in adult cardiovascular mortality, and are likely to be related to childhood environment(2). Blood pressure level in childhood has also been shown to track through to adulthood(3). Diet has been shown to relate to blood pressure(4) in childhood and there is evidence that dietary changes in very early life may have persistent effects on blood pressure(5). This suggests that dietary interventions aiming to lower blood pressure in childhood could have an impact on adult cardiovascular mortality. However, data on the associations between diet and blood pressure in childhood are limited. The aim of this study is to investigate the cross-sectional and longitudinal associations between dietary intakes and blood pressure levels measured in the CIF sample at ages 5 and 7 years, taking account of potential confounding variables. Key dietary variables considered will include intakes of sodium, potassium

, magnesium, calcium, fat, protein, wholegrains, fish, fruit and vegetables. These results will be of use in formulating dietary recommendations for children.

References

1. Stamler J, Stamler R, Neaton JD. Blood pressure, systolic and diastolic and cardiovascular risk factors: US population data. Archives of Internal Medicine. 1993;153:598-615.

2. Whincup PH, Cook DG, Adshead F, Taylor S, Papacosta O, Walker M, Wilson V. Cardiovascular risk factors in British children from towns with widely differing adult cardiovascular mortality. BMJ 1996;313:79-84.

3. Kivimaki M et al. Early socioeconomic position and blood pressure in childhood and adulthood: the Cardiovascular Risk in Young Finns Study. Hypertension 2006;47:39-44.

4. Simons-Morton DG et al. Nutrient intake and blood pressure in the Dietary Intervention Study in Children. Hypertension 1997;29:930-936.

5. Geleijnse JM, Hofman A, Witteman JCM et al., Long-term effects of neonatal sodium retention on blood pressure, Hypertension 1996; 29, pp. 913-917.

Data requested

1. Food and nutrient intakes assessed by dietary diary at 5 years and 7 years in the CIF sample

2. Blood pressure measured in the Focus@7 clinic in the CIF sample

3. Blood pressure measured at 5 year clinic in the CIF sample

4. Birthweight, measured or abstracted from medical records

5. Gestational age abstracted from medical records

6. Height and weight and waist circumference at 5 years measured in the CIF clinic

7. Height and weight and waist circumference measured in the Focus@7 clinic in the CIF sample

7. Physical fitness measured at 5 year clinic in the CIF sample

8. Measures of maternal educational level and paternal occupational class by questionnaire

9. Maternal smoking in pregnancy measured by questionnaire

10. Duration of breast-feeding assessed by questionnaire

11. Age of introduction of solids assessed by questionnaire

We want to us ALSPAC maths data as we feel that information on maths and maths teachers is particularly strong. Young people's attitudes to maths have been examined at different stages in their lives, and their ability has been tested through independent tests in years 4, 6 and 8 (covering specific domains of maths). In addition, there have been surveys of maths teachers' experience and attitudes in the main ALSPAC schools (in KS3) which could be linked to individual pupil data. None of this data has yet been analysed. Our understanding is that the data for assessments at Year 4 and Year 6 are now available, but Year 8 needs additional funding to clean and release the data, as does the data from the maths teacher questionnaires in 2002/3 and 2004/5, and the linking of questionnaires from teachers to children.

Once this data is available we intend to commission a separate analysis project. This will be competively tendered. The successful tenderer will require access to the data.

There are three possible areas for analysis:

* Young people's experience - using the questionnaires and independent assessments to examine how the young people have developed capability and confidence in maths, either looking at evidence throughout their lives (up to 14 or 16) or just at change between Y4-Y8.

* Teachers' experience - using the teachers' questionnaires to look at teacher attitudes and relating these to their school circumstances, stress and confidence

* Linking teacher and child experience - how teacher experience and attitudes link to child enjoyment, engagement and ability.

Funding may be available this financial year to make this data available. Please advise on timetable and costs.

This study will examine the nature of dating violence as reported by male and female adolescents and determine the applicability of the intergenerational hypothesis in each case. In addition, the study will examine the relationship between early socialization and the development of pro-violence attitudes.

There are three aims of this project:

1.The first is to provide a descriptive analysis of the incidence of dating violence within the ALSPAC cohort. This will include a gender based analysis of the use and receipt of violence within dating relationships as well as the young people's understanding of these behaviours and help seeking behaviours associated with the experience of these behaviours.

2. The second aim is to determine the extent to which the experience of violence within dating relationships (as both perpetrator and victim) reflects an intergenerational pattern. This will be examined in relation to the young person's possible exposure to inter-parental violence, harsh parental discipline and parental maltreatment, controlling for additional family adversity exposure and previous involvement in bullying and antisocial behaviour. Should the data permit, a gender based analysis of these issues will be conducted.

3. The final aim is to determine the extent to which pro-violence attitudes reflect earlier exposure to inter-parental violence, harsh parental discipline, parental maltreatment, after controlling for additional family adversity exposure and previous involvement in bullying, antisocial behaviour and dating violence.

As a consequence of this investigation at least three manuscripts will be written for submission in international peer reviewed journals. In addition, these preliminary investigations will provide the basis of continued research funding applications that will focus on examining potential mediators and moderators of any intergenerational effects wtihin a risk and resilience framework.

In order to fulfil these project aims, the following data is required:

Family Adversity Index (FAI) data with the maternal victimization variables isolated for separate analyses

Child Maltreatment Index - this will be derived from variables relating to maternal parenting practices.

Maternal reports of victimization

Paternal reports of victimization

Maternal parenting data

Paternal parenting data

Bullying involvement data

Dating violence data

Pro-violence attitude data

Details of the measures and observations are presented in table 1 (see attached).

Applications are invited from suitably qualified psychology graduates to join a team investigating the development of depressive symptoms. The PhD will be undertaken in the Department of Community Based Medicine and will focus on examining the nature of the relationship between depressive symptoms and smoking.

Previous studies have found evidence for an association between depression and smoking in adolescence, but the nature and direction of the relationship is unclear. Some studies have found evidence that depressive symptoms precede the onset of smoking; others have found that smoking precedes depression, and some studies report a bi-directional relationship. The PhD student will have the opportunity to develop skills in statistical modelling of longitudinal data to model the developmental heterogeneity of depression and smoking from late childhood into adolescence and examine potential covariates including gender, behaviour and conduct problems, social adversities, stressful life events, and parent-child relationships.

The project will take advantage of the unique and extensive longitudinal data collected by ALSPAC, an ongoing longitudinal population-based study investigating a wide range of environmental and other influences on the health and development of children. Detailed information on the ALSPAC study is available on the web site: http://www.alspac.bris.ac.uk.

Data required:

Depression- Mood and feelings questionnaire (child and parent report) from 9-15 years.

Smoking- (child and parent report) from 8-15 years.

(we are aware that data from the 15/16 year questionnaire/clinic is not yet available)

Other possible data requirements: Behaviour and emotional problems- DAWBA, SDQ, antisocial behaviour questionnaire.

Social adversities and stressful life events.

Questions on parent-child relationships.

The aim of the present study is to examine various victimization experiences (domestic, physical harshness) and peer victimization, and their association with BPD in a longitudinal study of a young general population sample with detailed assessment of traumatic events as well as BPD. The specific questions to be answered are: (1) What is the association between peer victimisation versus other traumatic victimisation in childhood and BPD at the of 11? (2) Is there a dose-response relationship between the number of victimisation events and the risk for BPD (3) Are the observed associations independent or mediated by other psychopathology?