Identification of plasma metabolites associated with modifiable risk factors and endophenotypes reflecting Alzheimer's disease pathology.

Modifiable factors can influence the risk for Alzheimer's disease (AD) and serve as targets for intervention; however, the biological mechanisms linking these factors to AD are unknown. This study aims to identify plasma metabolites associated with modifiable factors for AD, including MIND diet, physical activity, smoking, and caffeine intake, and test their association with AD endophenotypes to identify their potential roles in pathophysiological mechanisms. The association between each of the 757 plasma metabolites and four modifiable factors was tested in the wisconsin registry for Alzheimer's prevention cohort of initially cognitively unimpaired, asymptomatic middle-aged adults. After Bonferroni correction, the significant plasma metabolites were tested for association with each of the AD endophenotypes, including twelve cerebrospinal fluid (CSF) biomarkers, reflecting key pathophysiologies for AD, and four cognitive composite scores. Finally, causal mediation analyses were conducted to evaluate possible mediation effects. Analyses were performed using linear mixed-effects regression. A total of 27, 3, 23, and 24 metabolites were associated with MIND diet, physical activity, smoking, and caffeine intake, respectively. Potential mediation effects include beta-cryptoxanthin in the association between MIND diet and preclinical Alzheimer cognitive composite score, hippurate between MIND diet and immediate learning, glutamate between physical activity and CSF neurofilament light, and beta-cryptoxanthin between smoking and immediate learning. Our study identified several plasma metabolites that are associated with modifiable factors. These metabolites can be employed as biomarkers for tracking these factors, and they provide a potential biological pathway of how modifiable factors influence the human body and AD risk.

Large-scale proteome and metabolome analysis of CSF implicates altered glucose and carbon metabolism and succinylcarnitine in Alzheimer's disease.

INTRODUCTION: A hallmark of Alzheimer's disease (AD) is the aggregation of proteins (amyloid beta [A] and hyperphosphorylated tau [T]) in the brain, making cerebrospinal fluid (CSF) proteins of particular interest. METHODS: We conducted a CSF proteome-wide analysis among participants of varying AT pathology (n = 137 participants; 915 proteins) with nine CSF biomarkers of neurodegeneration and neuroinflammation. RESULTS: We identified 61 proteins significantly associated with the AT category (P < 5.46 × 10-5 ) and 636 significant protein-biomarker associations (P < 6.07 × 10-6 ). Proteins from glucose and carbon metabolism pathways were enriched among amyloid- and tau-associated proteins, including malate dehydrogenase and aldolase A, whose associations with tau were replicated in an independent cohort (n = 717). CSF metabolomics identified and replicated an association of succinylcarnitine with phosphorylated tau and other biomarkers. DISCUSSION: These results implicate glucose and carbon metabolic dysregulation and increased CSF succinylcarnitine levels with amyloid and tau pathology in AD. HIGHLIGHTS: Cerebrospinal fluid (CSF) proteome enriched for extracellular, neuronal, immune, and protein processing. Glucose/carbon metabolic pathways enriched among amyloid/tau-associated proteins. Key glucose/carbon metabolism protein associations independently replicated. CSF proteome outperformed other omics data in predicting amyloid/tau positivity. CSF metabolomics identified and replicated a succinylcarnitine-phosphorylated tau association.

Interaction of amyloid and tau on cortical microstructure in cognitively unimpaired adults.

INTRODUCTION: Neurite orientation dispersion and density imaging (NODDI), a multi-compartment diffusion-weighted imaging (DWI) model, may be useful for detecting early cortical microstructural alterations in Alzheimer's disease prior to cognitive impairment. METHODS: Using neuroimaging (NODDI and T1-weighted magnetic resonance imaging [MRI]) and cerebrospinal fluid (CSF) biomarker data (measured using Elecsys® CSF immunoassays) from 219 cognitively unimpaired participants, we tested the main and interactive effects of CSF amyloid beta (Aß)42 /Aß40 and phosphorylated tau (p-tau) on cortical NODDI metrics and cortical thickness, controlling for age, sex, and apolipoprotein E ε4. RESULTS: We observed a significant CSF Aß42 /Aß40 × p-tau interaction on cortical neurite density index (NDI), but not orientation dispersion index or cortical thickness. The directionality of these interactive effects indicated: (1) among individuals with lower CSF p-tau, greater amyloid burden was associated with higher cortical NDI; and (2) individuals with greater amyloid and p-tau burden had lower cortical NDI, consistent with cortical neurodegenerative changes. DISCUSSION: NDI is a particularly sensitive marker for early cortical changes that occur prior to gross atrophy or development of cognitive impairment.

Longitudinal effects of family psychopathology and stress on pubertal maturation and hormone coupling in adolescent twins.

Adolescents experience profound neuroendocrine changes, including hormone "coupling" between cortisol, testosterone, and dehydroepiandrosterone. Emerging research has only begun to elucidate the role of hormone coupling, its genetic and environmental etiology, and the extent to which coupling is impacted by gender, puberty, and family context. We included measures on parent and child mental health, parenting stress, and family conflict of 444 twin pairs and their parents across two timepoints, when youth were on average 8 and 13 years old, respectively. Structural equation models examined the impact of family context effects on coupling during adolescence. Biometric twin models were then used to probe additive genetic, shared, and non-shared environmental effects on hormone coupling. Hormones were more tightly coupled for females than males, and coupling was sensitive to parental depression and co-twin psychopathology symptoms and stress exposure in females. The association between family context and coupling varied across specific neuroendocrine measures and was largely distinct from pubertal maturation. Biometric models revealed robust shared and non-shared environmental influences on coupling. We found that family antecedents modify the strength of coupling. Environmental influences account for much of the variation on coupling during puberty. Gender differences were found in genetic influences on coupling.

Attentional Control Explains Covariation Between Symptoms of Attention-Deficit/Hyperactivity Disorder and Anxiety During Adolescence.

Symptoms of attention-deficit/hyperactivity disorder (ADHD) and anxiety are common during adolescence and frequently co-occur. However, the genetic and environmental influences that underlie this co-occurrence are understudied. Using a large twin sample (N = 1,017), we examined cross-sectional genetic and environmental influences on ADHD and anxiety symptoms during childhood. We also explored whether these influences were shared with attentional control, a putative mechanism for symptom comorbidity. We found evidence for common genetic and nonshared environmental influences on the covariation among attentional control, ADHD, and anxiety symptoms, supporting the putative role of attentional control as a mechanism by which comorbid problems may develop. Genetic factors also accounted for symptom co-occurrence after controlling for covariation with attentional control, suggesting the presence of additional unmeasured mechanisms.

Longitudinal Research at the Interface of Affective Neuroscience, Developmental Psychopathology, Health and Behavioral Genetics: Findings from the Wisconsin Twin Project.

The Wisconsin Twin Project comprises multiple longitudinal studies that span infancy to early adulthood. We summarize recent papers that show how twin designs with deep phenotyping, including biological measures, can inform questions about phenotypic structure, etiology, comorbidity, heterogeneity, and gene-environment interplay of temperamental constructs and mental and physical health conditions of children and adolescents. The general framework for investigations begins with rich characterization of early temperament and follows with study of experiences and exposures across childhood and adolescence. Many studies incorporate neuroimaging and hormone assays.

Sex Differences in the Genetic and Environmental Influences on Self-Reported Non-aggressive and Aggressive Conduct Disorder Symptoms in Early and Middle Adolescence.

Decades of research into the etiology of conduct disorder (CD) has yet to yield a consensus on the existence of sex differences in underlying genetic and environmental influences. This may be partly due to the failure of many previous studies to make a distinction between non-aggressive and aggressive CD symptoms or test for potential developmental changes in sex differences in the etiology of conduct problems. To address these gaps, we fit a series of univariate and bivariate biometric sex-difference models to self-reported non-aggressive and aggressive CD symptoms in a community-based sample of twins (N = 1548, ages 9-17 year), grouped into ages 9-13 and 14-17 years. Relative model fit was evaluated using the Bayesian Information Criterion (BIC), which favors parsimony, and by Chi square difference tests. The univariate sex-scalar model provided the best fit to the data for both non-aggressive and aggressive CD symptoms at ages 9-13 and 14-17 years. Thus, the same genetic and environmental factors influenced CD symptoms in both sexes, but the total variability was lower in females than males. At both ages, the heritability of non-aggressive CD symptoms was lower than heritability of aggressive CD symptoms, and shared environmental effects were only observed for non-aggressive CD symptoms. However, estimates for genetic and environmental factors could be not be constrained to be equal across age groups for either CD subtype, suggesting substantive developmental changes in the relative influence of genetic and environmental factors on individual differences in CD symptoms. For both subtypes, the heritability was larger, and shared environmental effect smaller, in the older age group than the younger age group. A bivariate quantitative sex differences model provided the best fit to the data at ages 9-13 years. Covariation between non-aggressive and aggressive CD symptoms was due to overlapping shared and non-shared environmental factors in males and females but the overall covariation was greater in males than females. In contrast, at ages 14-17 years, the sex-scalar bivariate model provided the best fit to the data, and covariation between non-aggressive and aggressive CD symptoms was due to overlapping genetic and non-shared environmental factors. Thus, the etiology of self-reported conduct disorder varied substantially by symptom type and age. However, quantitative sex differences were only apparent when the covariation between the two subtypes was considered.

Infant stranger fear trajectories predict anxious behaviors and diurnal cortisol rhythm during childhood.

Although a robust literature has linked stable, high levels of fear across childhood to increased risk for anxiety problems, less is known about alternative pathways to anxiety. We tested two putatively normative developmental pathways of early fearfulness for their distinct associations with behavioral (anxiety-related behaviors and symptoms) and biological (diurnal cortisol) markers of anxiety risk in middle childhood in a community-based sample (n = 107). Steeper increases in fear from 6 to 36 months predicted more parent-reported anxiety symptoms at age 8 years. In addition, children who exhibited steep increases in fear during infancy were overrepresented among children with diagnoses of separation anxiety disorder at age 8 years. Finally, we showed that steeper increases in fearfulness in infancy predicted flatter slopes of diurnal cortisol at age 8 years for girls. Thus, differences in stranger fear across infancy may indicate varying degrees of risk for anxious behaviors in later childhood.

Partial replication of two rumination-related candidate gene studies.

Two recent papers associated candidate genes with brooding rumination, a possible cognitive endophenotype for depression, in children ages 8-14 years. Stone et al. reported that BDNF val66met polymorphism predicted brooding in adolescence. Woody et al. reported that children carrying at least one copy of a CRHR1 TAT haplotype reported less brooding than their peers in the presence of maternal depression. We attempted to replicate and extend these findings in a sample of twins aged 12-16 years. We analyzed the BDNF val66met (rs6265) polymorphism and two (rs242924 and rs7209436) out of three single nucleotide polymorphisms (SNPs) that Woody et al. used to create a CRHR1 haplotype. We controlled for maternal history of depression and clustering within families. Unlike Stone et al., we found higher brooding among BDNF Met carriers. This main effect was qualified by an interaction with pubertal status, with the effect driven by more physically mature participants. Similar to Woody et al., we found an interaction between CRHR1 SNPs and maternal depression, with the homozygous minor genotype acting as a protective factor against brooding in the presence of maternal depression. Findings provide partial support for the influence of candidate genes in two environmentally sensitive systems on brooding.

Genetic and Environmental Contributions to Associations between Infant Fussy Temperament and Antisocial Behavior in Childhood and Adolescence.

Previous research suggests that fussy temperament in infancy predicts risk for later antisocial behavior (ASB) in childhood and adolescence. It remains unclear, however, to what extent infant fussiness is related to later ASB through causal processes or if they both reflect the same family risk factors for ASB. The current study used two approaches, the comparison of siblings and bivariate biometric modeling, to reduce familial confounding and examine genetic and environmental influences on associations between fussiness in the first 2 years of life and ASB in childhood and late adolescence. Analyses were conducted on data from a prospective cohort (9237 at 4-9 years and 7034 at 14-17 years) who are the offspring of a nationally representative sample of US women. In the full sample, fussiness predicted both child and adolescent ASB to small but significant extents, controlling for a wide range of measured child and family-level covariates. When siblings who differed in their fussiness were compared, fussiness predicted ASB in childhood, but not ASB during adolescence. Furthermore, results from a bivariate Cholesky model suggested that even the association of fussiness with childhood ASB found when comparing siblings is attributable to familial factors. That is, although families with infants who are higher in fussiness also tend to have children and adolescents who engage in greater ASB, the hypothesis that infant fussiness has an environmentally mediated impact on the development of future ASB was not strongly supported.

Comparing Alternative Biometric Models with and without Gene-by-Measured Environment Interaction in Behavior Genetic Designs: Statistical Operating Characteristics.

To extend Purcell's well known ACE model in testing gene by measured environment interactions (GxM) in behavior genetic designs, Rathouz et al. considered a broader class of models for quantifying and testing such interactions. Only a sub-group of these extended models have been investigated for their statistical operating characteristics by Van Hulle et al. due to lack of closed form likelihood. With an estimation procedure developed using numerical techniques in a companion paper, we study statistical operating characteristics of these extended models, especially those with non-linear effects. Type I error analysis shows the likelihood ratio test for GxM to be conservative in testing models extended from the bivariate Cholesky model, and to be liberal for models extended from the bivariate correlated factors model. Parameter estimation for all models is very good, with little bias exhibited for most models and parameters. Comparisons among alternative models under various simulated conditions show that it is relatively more difficult to confirm the existence of gene by environment interactions versus to detect non-linear effects which exclude such interactions.

Genetic and Environmental Contributions to Covariation Between DHEA and Testosterone in Adolescent Twins.

Although several studies have shown that pubertal tempo and timing are shaped by genetic and environmental factors, few studies consider to what extent endocrine triggers of puberty are shaped by genetic and environmental factors. Doing so moves the field from examining correlated developmentally-sensitive biomarkers toward understanding what drives those associations. Two puberty related hormones, dehydroepiandrosterone and testosterone, were assayed from salivary samples in 118 MZ (62 % female), 111 same sex DZ (46 % female) and 103 opposite-sex DZ twin pairs, aged 12-16 years (M = 13.1, SD = 1.3). Pubertal status was assessed with a composite of mother- and self-reports. We used biometric models to estimate the genetic and environmental influences on the variance and covariance in testosterone and DHEA, with and without controlling for their association with puberty, and to test for sex differences. In males, the variance in testosterone and pubertal status was due to shared and non-shared environmental factors; variation in DHEA was due to genetic and non-shared environmental factors. In females, variance in testosterone was due to genetic and non-shared environmental factors; genetic, shared, and non-shared environmental factors contributed equally to variation in DHEA. In males, the testosterone-DHEA covariance was primarily due to shared environmental factors that overlapped with puberty as well as shared and non-shared environmental covariation specific to testosterone and DHEA. In females, the testosterone-DHEA covariance was due to genetic factors overlapping with pubertal status, and shared and non-shared environmental covariation specific to testosterone and DHEA.

Operating Characteristics of Statistical Methods for Detecting Gene-by-Measured Environment Interaction in the Presence of Gene-Environment Correlation under Violations of Distributional Assumptions.

Accurately identifying interactions between genetic vulnerabilities and environmental factors is of critical importance for genetic research on health and behavior. In the previous work of Van Hulle et al. (Behavior Genetics, Vol. 43, 2013, pp. 71-84), we explored the operating characteristics for a set of biometric (e.g., twin) models of Rathouz et al. (Behavior Genetics, Vol. 38, 2008, pp. 301-315), for testing gene-by-measured environment interaction (GxM) in the presence of gene-by-measured environment correlation (rGM) where data followed the assumed distributional structure. Here we explore the effects that violating distributional assumptions have on the operating characteristics of these same models even when structural model assumptions are correct. We simulated N = 2,000 replicates of n = 1,000 twin pairs under a number of conditions. Non-normality was imposed on either the putative moderator or on the ultimate outcome by ordinalizing or censoring the data. We examined the empirical Type I error rates and compared Bayesian information criterion (BIC) values. In general, non-normality in the putative moderator had little impact on the Type I error rates or BIC comparisons. In contrast, non-normality in the outcome was often mistaken for or masked GxM, especially when the outcome data were censored.

Clarifying the associations between age at menarche and adolescent emotional and behavioral problems.

Better understanding risk factors for the development of adolescent emotional and behavioral problems can help with intervention and prevention efforts. Previous studies have found that an early menarcheal age predicts several adolescent problems, including depressive symptoms, delinquency, and early age at first intercourse. Few studies, nevertheless, have explicitly tested (a) whether the associations with menarcheal age vary across racial/ethnic groups or (b) whether the sources of the associations are within-families (i.e., consistent with a direct, causal link) or only between-families (i.e., due to selection or confounding factors). The current study analyzed data from a nationally representative US Sample of females (N = 5,637). We examined whether race/ethnicity moderated the associations between early menarche and several adolescent problems by using multiple-group analyses and we examined the degree to which genetic and environmental factors shared by family members account for the associations by comparing sisters and cousins with differing menarcheal ages. Menarcheal age predicted subsequent depressive symptoms, delinquency, and early age at first intercourse in the population. The magnitudes of the associations were similar across all racial/ethnic groups for all outcomes. The within-family associations (i.e., when comparing siblings and cousins with different menarcheal age) were large and statistically significant when predicting early intercourse, but not the other outcomes. The findings suggest that selection or confounding factors account for the associations between menarcheal age and subsequent depressive symptoms and delinquency, whereas the independent association between menarcheal age and early age at first intercourse is consistent with a direct, causal effect.

A sibling-comparison study of smoking during pregnancy and childhood psychological traits.

Prenatal exposure to substances of abuse is associated with numerous psychological problems in offspring, but quasi-experimental studies controlling for co-occurring risk factors suggest that familial factors (e.g., genetic and environmental effects shared among siblings) confound many associations with maternal smoking during pregnancy (SDP). Few of the quasi-experimental studies in this area have explored normative psychological traits in early childhood or developmental changes across the lifespan, however. The current study used multilevel growth curve models with a large, nationally-representative sample in the United States to investigate for potential effects of SDP on the developmental trajectories of cognitive functioning, temperament/personality, and disruptive behavior across childhood, while accounting for shared familial confounds by comparing differentially exposed siblings and statistically controlling for offspring-specific covariates. Maternal SDP predicted the intercept (but not change over time) for all cognitive and externalizing outcomes. Accounting for familial confounds, however, attenuated the association between SDP exposure and all outcomes, except the intercept (age 5) for reading recognition. These findings, which are commensurate with previous quasi-experimental research on more severe indices of adolescent and adult problems, suggest that the associations between SDP and developmental traits in childhood are due primarily to confounding factors and not a causal association.

Ethnoracialized group differences in attitudes and knowledge about schizophrenia and willingness to engage in biomarker research: The UBIGR Study.

Although there is renewed optimism in biomarker research in schizophrenia, there is also need for greater inclusion of historically underrepresented groups in the research. In the present study, we surveyed 599 African American, 352 American Indian/Alaska Native, and 725 NonHispanic White participants about their attitudes toward research, knowledge and attitudes about schizophrenia, and willingness to engage in biomarker testing. Attitudes toward research were examined using the standardized 7-item Research Attitudes Questionnaire (RAQ) measure. Using structural equation modeling (SEM), we tested our predictive model of the likelihood of willingness to engage in biomarker testing for schizophrenia risk. Members of historically underrepresented groups were less willing to engage in biomarker testing. Overall, attitudes toward research, particularly trust, influenced biomarker testing willingness. These findings suggest that factors influencing willingness to engage in schizophrenia biomarker testing may be modifiable by outreach engagement and education.

Associations between recall of proper names in story recall and CSF amyloid and tau in adults without cognitive impairment.

Neuropsychological measures sensitive to decline in the preclinical phase of Alzheimer's disease are needed. We previously demonstrated that higher amyloid-beta (Aß) assessed by positron emission tomography in adults without cognitive impairment was associated with recall of fewer proper names in Logical Memory story recall. The current study investigated the association between proper names and cerebrospinal fluid biomarkers (Aß42/40, phosphorylated tau181 [pTau181], neurofilament light) in 223 participants from the Wisconsin Registry for Alzheimer's Prevention. We assessed associations between biomarkers and delayed Logical Memory total score and proper names using binary logistic regressions. Sensitivity analyses used multinomial logistic regression and stratified biomarker groups. Lower Logical Memory total score and proper names scores from the most recent visit were associated with biomarker positivity. Relatedly, there was a 27% decreased risk of being classified Aß42/40+/pTau181+ for each additional proper name recalled. A linear mixed effects model found that longitudinal change in proper names recall was predicted by biomarker status. These results demonstrate a novel relationship between proper names and Alzheimer's disease-cerebrospinal fluid pathology.

Operating characteristics of alternative statistical methods for detecting gene-by-measured environment interaction in the presence of gene-environment correlation in twin and sibling studies.

It is likely that all complex behaviors and diseases result from interactions between genetic vulnerabilities and environmental factors. Accurately identifying such gene-environment interactions is of critical importance for genetic research on health and behavior. In a previous article we proposed a set of models for testing alternative relationships between a phenotype (P) and a putative moderator (M) in twin studies. These include the traditional bivariate Cholesky model, an extension of that model that allows for interactions between M and the underling influences on P, and a model in which M has a non-linear main effect on P. Here we use simulations to evaluate the type I error rates, power, and performance of the Bayesian Information Criterion under a variety of data generating mechanisms and samples sizes (n = 2,000 and n = 500 twin pairs). In testing the extension of the Cholesky model, false positive rates consistently fell short of the nominal Type I error rates ([Formula: see text]). With adequate sample size (n = 2,000 pairs), the correct model had the lowest BIC value in nearly all simulated datasets. With lower sample sizes, models specifying non-linear main effects were more difficult to distinguish from models containing interaction effects. In addition, we provide an illustration of our approach by examining possible interactions between birthweight and the genetic and environmental influences on child and adolescent anxiety using previously collected data. We found a significant interaction between birthweight and the genetic and environmental influences on anxiety. However, the interaction was accounted for by non-linear main effects of birthweight on anxiety, verifying that interaction effects need to be tested against alternative models.

Early concern and disregard for others as predictors of antisocial behavior.

BACKGROUND: Prediction of antisocial behavior is important, given its adverse impact on both the individuals engaging in antisocial behavior and society. Additional research identifying early predictors of future antisocial behavior, or antisocial propensity, is needed. The present study tested the hypothesis that both concern for others and active disregard for others in distress in toddlers and young children predict antisocial behavior during middle childhood and adolescence. METHODS: A representative sample of same-sex twins (N=956) recruited in Colorado was examined. Mother-rated and researcher-observed concern and disregard for others assessed at age 14-36 months were examined as predictors of parent- (age 4-12), teacher- (age 7-12), and self-reported (age 17) antisocial behavior. RESULTS: Observed disregard for others predicted antisocial behavior assessed by three different informants (parents, teachers, and self), including antisocial behavior assessed 14 years later. It also predicted a higher order antisocial behavior factor (ß=.58, p<.01) after controlling for observed concern for others. Mother-rated disregard for others predicted parent-reported antisocial behavior. Contrary to predictions, neither mother-rated nor observed concern for others inversely predicted antisocial behavior. RESULTS of twin analyses suggested that the covariation between observed disregard for others and antisocial behavior was due to shared environmental influences. CONCLUSIONS: Disregard for others in toddlerhood/early childhood is a strong predictor of antisocial behavior in middle childhood and adolescence. The results suggest the potential need for early assessment of disregard for others and the development of potential interventions.