Computationally efficient familywise error rate control in genome-wide association studies using score tests for generalized linear models

In genetic association studies, detecting phenotype–genotype association is a primary goal. We assume that the relationship between the data—phenotype, genetic markers and environmental covariates—can be modeled by a generalized linear model. The number of markers is allowed to be far greater than the number of individuals of the study. A multivariate score statistic is used to test each marker for association with a phenotype. We assume that the test statistics asymptotically follow a multivariate normal distribution under the complete null hypothesis of no phenotype–genotype association. We present the familywise error rate order k approximation method to find a local significance level (alternatively, an adjusted p-value) for each test such that the familywise error rate is controlled. The special case k=1 gives the Šidák method. As a by-product, an effective number of independent tests can be defined. Furthermore, if environmental covariates and genetic markers are uncorrelated, or no environmental covariates are present, we show that covariances between score statistics depend on genetic markers alone. This not only leads to more efficient calculations but also to a local significance level that is determined only by the collection of markers used, independent of the phenotypes and environmental covariates of the experiment at hand.     

Empirical likelihood-based robust tests for genetic association analysis with quantitative traits

Genome-wide association studies (GWAS) are effective in investigating the loci related with complex diseases. For most of these studies, the genetic inheritance model is not known in advance and therefore robust tests are preferred. Empirical likelihood (EL) method is well known for its flexibility and nonparametric properties, but is rarely investigated in GWAS. In this study, we develop EL-based test statistics to detect the association of a disease and genetic loci while the genetic model is unknown. The performance of proposed tests is evaluated by simulations and compared with several existing methods. For illustration, we apply these tests to identify the single nucleotide polymorphisms associated with alkaline phosphatase level on mouse chromosome 6.     

Choosing between per-genotype,per-allele,and trend approaches for initial detection of gene–disease association

There are a number of approaches to detect candidate gene–disease associations including: (i) ‘per-genotype’, which looks for any difference across the genotype groups without making any assumptions about the direction of the effect or the genetic model; (ii) ‘per-allele’, which assumes an additive genetic model, i.e. an effect for each allele copy; and (iii) linear trend, which looks for an incremental effect across the genotype groups. We simulated a number of gene–disease associations, varying odds ratios, allele frequency, genetic model, and deviation from Hardy–Weinberg equilibrium (HWE) and tested the performance of each of the three methods to detect the associations, where performance was judged by looking at critical values, power, coverage, bias, and root mean square error. Results indicate that the per-allele method is very susceptible to false positives and false negatives when deviations from HWE occur. The linear trend test appears to have the best power under most simulated scenarios, but can sometimes be biased and have poor coverage. These results indicate that of these strategies a linear trend test may be best for initially testing an association, and the per-genotype approach may be best for estimating the magnitude of the association.     

Bayesian hierarchical regression models for detecting QTLs in plant experiments

Quantitative trait loci (QTL) mapping is a growing field in statistical genetics. In plants, QTL detection experiments often feature replicates or clones within a specific genetic line. In this work, a Bayesian hierarchical regression model is applied to simulated QTL data and to a dataset from the Arabidopsis thaliana plants for locating the QTL mapping associated with cotyledon opening. A conditional model search strategy based on Bayesian model averaging is utilized to reduce the computational burden.     

Candidate-gene association analysis for a continuous phenotype with a spike at zero using parent-offspring trios

In this paper, we propose the class of generalized additive models for location, scale and shape in a test for the association of genetic markers with non-normally distributed phenotypes comprising a spike at zero. The resulting statistical test is a generalization of the quantitative transmission disequilibrium test with mating type indicator, which was originally designed for normally distributed quantitative traits and parent-offspring data. As a motivational example, we consider coronary artery calcification (CAC), which can accurately be identified by electron beam tomography. In the investigated regions, individuals will have a continuous measure of the extent of calcium found or they will be calcium-free. Hence, the resulting distribution is a mixed discrete-continuous distribution with spike at zero. We carry out parent-offspring simulations motivated by such CAC measurement values in a screening population to study statistical properties of the proposed test for genetic association. Furthermore, we apply the approach to data of the Genetic Analysis Workshop 16 that are based on real genotype and family data of the Framingham Heart Study, and test the association of selected genetic markers with simulated coronary artery calcification.     

Case–control genome-wide joint association study using semiparametric empirical model and approximate Bayes factor

We propose a semiparametric approach for the analysis of case–control genome-wide association study. Parametric components are used to model both the conditional distribution of the case status given the covariates and the distribution of genotype counts, whereas the distribution of the covariates are modelled nonparametrically. This yields a direct and joint modelling of the case status, covariates and genotype counts, and gives a better understanding of the disease mechanism and results in more reliable conclusions. Side information, such as the disease prevalence, can be conveniently incorporated into the model by an empirical likelihood approach and leads to more efficient estimates and a powerful test in the detection of disease-associated SNPs. Profiling is used to eliminate a nuisance nonparametric component, and the resulting profile empirical likelihood estimates are shown to be consistent and asymptotically normal. For the hypothesis test on disease association, we apply the approximate Bayes factor (ABF) which is computationally simple and most desirable in genome-wide association studies where hundreds of thousands to a million genetic markers are tested. We treat the approximate Bayes factor as a hybrid Bayes factor which replaces the full data by the maximum likelihood estimates of the parameters of interest in the full model and derive it under a general setting. The deviation from Hardy–Weinberg Equilibrium (HWE) is also taken into account and the ABF for HWE using cases is shown to provide evidence of association between a disease and a genetic marker. Simulation studies and an application are further provided to illustrate the utility of the proposed methodology.     

Analysis of ordered categorical data with score averaging: with applications to case-control genetic associations

The trend test is often used for the analysis of 2×K ordered categorical data, in which K pre-specified increasing scores are used. There have been discussions on how to assign these scores and the impact of the outcomes on different scores. The scores are often assigned based on the data-generating model. When this model is unknown, using the trend test is not robust. We discuss the weighted average of a trend test over all scientifically plausible choices of scores or models. This approach is more computationally efficient than a commonly used robust test MAX when K is large. Our discussion is for any ordered 2×K table, but simulation and applications to real data are focused on case-control genetic association studies. Although there is no single test optimal for all choices of scores, our numerical results show that some score averaging tests can achieve the performance of MAX.     

The power of generalized odds ratio in assessing association in genetic studies with known mode of inheritance

The generalized odds ratio (OR_G) is a novel model-free approach to test the association in genetic studies by estimating the overall risk effect based on the complete genotype distribution. However, the power of OR_G has not been explored and, particularly, in a setting where the mode of inheritance is known. A population genetics model was simulated in order to define the mode of inheritance of a pertinent gene–disease association in advance. Then, the power of OR_G was explored based on this model and compared with the chi-square test for trend. The model considered bi- and tri-allelic gene–disease associations, and deviations from the Hardy–Weinberg equilibrium (HWE). The simulations showed that bi- and tri-allelic variants have the same pattern of power results. The power of OR_G increases with increase in the frequency of mutant allele and the coefficient of selection and, of course, the degree of dominance of the mutant allele. The deviation from HWE has a considerable impact on power only for small values of the above parameters. The OR_G showed superiority in power compared with the chi-square test for trend when there is deviation from HWE; otherwise, the pattern of results was similar in both the approaches.     

SNP selection for predicting a quantitative trait

Molecular markers combined with powerful statistical tools have made it possible to detect and analyze multiple loci on the genome that are responsible for the phenotypic variation in quantitative traits. The objectives of the study presented in this paper are to identify a subset of single nucleotide polymorphism (SNP) markers that are associated with a particular trait and to construct a model that can best predict the value of the trait given the genotypic information of the SNPs using a three-step strategy. In the first step, a genome-wide association test is performed to screen SNPs that are associated with the quantitative trait of interest. SNPs with p-values of less than 5% are then analyzed in the second step. In the second step, a large number of randomly selected models, each consisting of a fixed number of randomly selected SNPs, are analyzed using the least angle regression method. This step will further remove redundant SNPs due to the complicated association among SNPs. A subset of SNPs that are shown to have a significant effect on the response trait more often than by chance are considered for the third step. In the third step, two alternative methods are considered: the least angle shrinkage and selection operation and sparse partial least squares regression. For both methods, the predictive ability of the fitted model is evaluated by an independent test set. The performance of the proposed method is illustrated by the analysis of a real data set on Canadian Holstein cattle.     

Robust suptest for the genetic association study under genetic model uncertainty

In case–control studies the Cochran–Armitage trend test is powerful for detection of an association between a risk genetic marker and a disease of interest. To apply this test, a score should be assigned to the genotypes based on the genetic model. When the underlying genetic model is unknown, the trend test statistic is quite sensitive to the choice of the score. In this paper, we study the asymptotic property of the robust suptest statistic defined as a supremum of Cochran–Armitage trend test across all scores between 0 and 1. Through numerical studies we show that small to moderate sample size performances of the suptest appear reasonable in terms of type I error control and we compared empirical powers of the suptest to those of three individual Cochran–Armitage trend tests and the maximum of the three Cochran–Armitage trend tests. The use of the suptest is applied to rheumatoid arthritis data from a genome-wide association study.     

Multivariate association test for rare variants controlling for cryptic and family relatedness

In genetic studies of complex diseases, multiple measures of related phenotypes are often collected. Jointly analyzing these phenotypes may improve statistical power to detect sets of rare variants affecting multiple traits. In this work, we consider association testing between a set of rare variants and multiple phenotypes in family‐based designs. We use a mixed linear model to express the correlations among the phenotypes and between related individuals. Given the many sources of correlations in this situation, deriving an appropriate test statistic is not straightforward. We derive a vector of score statistics, whose joint distribution is approximated using a copula. This allows us to have closed‐form expressions for the p‐values of several test statistics. A comprehensive simulation study and an application to Genetic Analysis Workshop 18 data highlight the gains associated with joint testing over univariate approaches, especially in the presence of pleiotropy or highly correlated phenotypes. The Canadian Journal of Statistics 47: 90–107; 2019 © 2018 Statistical Society of Canada     

Some applications of genetics in statistical ecology

Genetic data are in widespread use in ecological research, and an understanding of this type of data and its uses and interpretations will soon be an imperative for ecological statisticians. Here, we provide an introduction to the subject, intended for statisticians who have no previous knowledge of genetics. Although there are numerous types of genetic data, we restrict attention to multilocus genotype data from microsatellite loci. We look at two application areas in wide use: investigating population structure using genetic assignment and related techniques; and using genotype data in capture–recapture studies for estimating population size and demographic parameters. In each case, we outline the conceptual framework and draw attention to both the strengths and weaknesses of existing approaches to analysis and interpretation.     

CAUSAL GRAPHICAL MODELS IN SYSTEMS GENETICS: A UNIFIED FRAMEWORK FOR JOINT INFERENCE OF CAUSAL NETWORK AND GENETIC ARCHITECTURE FOR CORRELATED PHENOTYPES

Causal inference approaches in systems genetics exploit quantitative trait loci (QTL) genotypes to infer causal relationships among phenotypes. The genetic architecture of each phenotype may be complex, and poorly estimated genetic architectures may compromise the inference of causal relationships among phenotypes. Existing methods assume QTLs are known or inferred without regard to the phenotype network structure. In this paper we develop a QTL-driven phenotype network method (QTLnet) to jointly infer a causal phenotype network and associated genetic architecture for sets of correlated phenotypes. Randomization of alleles during meiosis and the unidirectional influence of genotype on phenotype allow the inference of QTLs causal to phenotypes. Causal relationships among phenotypes can be inferred using these QTL nodes, enabling us to distinguish among phenotype networks that would otherwise be distribution equivalent. We jointly model phenotypes and QTLs using homogeneous conditional Gaussian regression models, and we derive a graphical criterion for distribution equivalence. We validate the QTLnet approach in a simulation study. Finally, we illustrate with simulated data and a real example how QTLnet can be used to infer both direct and indirect effects of QTLs and phenotypes that co-map to a genomic region.     

Correcting for Population Stratification in Genomewide Association Studies

Genomewide association studies have become the primary tool for discovering the genetic basis of complex human diseases. Such studies are susceptible to the confounding effects of population stratification, in that the combination of allele-frequency heterogeneity with disease-risk heterogeneity among different ancestral subpopulations can induce spurious associations between genetic variants and disease. This article provides a statistically rigorous and computationally feasible solution to this challenging problem of unmeasured confounders. We show that the odds ratio of disease with a genetic variant is identifiable if and only if the genotype is independent of the unknown population substructure conditional on a set of observed ancestry-informative markers in the disease-free population. Under this condition, the odds ratio of interest can be estimated by fitting a semiparametric logistic regression model with an arbitrary function of a propensity score relating the genotype probability to ancestry-informative markers. Approximating the unknown function of the propensity score by B-splines, we derive a consistent and asymptotically normal estimator for the odds ratio of interest with a consistent variance estimator. Simulation studies demonstrate that the proposed inference procedures perform well in realistic settings. An application to the well-known Wellcome Trust Case-Control Study is presented. Supplemental materials are available online.     

Mixture cure model with an application to interval mapping of quantitative trait loci

When censored time-to-event data are used to map quantitative trait loci (QTL), the existence of nonsusceptible subjects entails extra challenges. If the heterogeneous susceptibility is ignored or inappropriately handled, we may either fail to detect the responsible genetic factors or find spuriously significant locations. In this article, an interval mapping method based on parametric mixture cure models is proposed, which takes into consideration of nonsusceptible subjects. The proposed model can be used to detect the QTL that are responsible for differential susceptibility and/or time-to-event trait distribution. In particular, we propose a likelihood-based testing procedure with genome-wide significance levels calculated using a resampling method. The performance of the proposed method and the importance of considering the heterogeneous susceptibility are demonstrated by simulation studies and an application to survival data from an experiment on mice infected with Listeria monocytogenes.     

Large sample interval mapping method for genetic trait loci in finite regression mixture models

This article investigates the large sample interval mapping method for genetic trait loci (GTL) in a finite non-linear regression mixture model. The general model includes most commonly used kernel functions, such as exponential family mixture, logistic regression mixture and generalized linear mixture models, as special cases. The populations derived from either the backcross or intercross design are considered. In particular, unlike all existing results in the literature in the finite mixture models, the large sample results presented in this paper do not require the boundness condition on the parametric space. Therefore, the large sample theory presented in this article possesses general applicability to the interval mapping method of GTL in genetic research. The limiting null distribution of the likelihood ratio test statistics can be utilized easily to determine the threshold values or p-values required in the interval mapping. The limiting distribution is proved to be free of the parameter values of null model and free of the choice of a kernel function. Extension to the multiple marker interval GTL detection is also discussed. Simulation study results show favorable performance of the asymptotic procedure when sample sizes are moderate.     

Some Statistical Properties of Efficiency Robust Tests with Applications to Genetic Association Studies

Although efficiency robust tests are preferred for genetic association studies when the genetic model is unknown, their statistical properties have been studied for different study designs separately under special situations. We study some statistical properties of the maximin efficiency robust test and a maximum‐type robust test (MAX3) under a general setting and obtain unified results. The results can also be applied to testing hypothesis with a constrained two‐dimensional parameter space. The results are applied to genetic association studies using case–parents trio data.     

USING LINEAR PREDICTORS TO IMPUTE ALLELE FREQUENCIES FROM SUMMARY OR POOLED GENOTYPE DATA

Recently-developed genotype imputation methods are a powerful tool for detecting untyped genetic variants that affect disease susceptibility in genetic association studies. However, existing imputation methods require individual-level genotype data, whereas in practice it is often the case that only summary data are available. For example this may occur because, for reasons of privacy or politics, only summary data are made available to the research community at large; or because only summary data are collected, as in DNA pooling experiments. In this article, we introduce a new statistical method that can accurately infer the frequencies of untyped genetic variants in these settings, and indeed substantially improve frequency estimates at typed variants in pooling experiments where observations are noisy. Our approach, which predicts each allele frequency using a linear combination of observed frequencies, is statistically straight-forward, and related to a long history of the use of linear methods for estimating missing values (e.g. Kriging). The main statistical novelty is our approach to regularizing the covariance matrix estimates, and the resulting linear predictors, which is based on methods from population genetics. We find that, besides being both fast and flexible - allowing new problems to be tackled that cannot be handled by existing imputation approaches purpose-built for the genetic context - these linear methods are also very accurate. Indeed, imputation accuracy using this approach is similar to that obtained by state-of-the art imputation methods that use individual-level data, but at a fraction of the computational cost.     

Testing for state dependence in binary panel data with individual covariates by a modified quadratic exponential model

We propose a test for state dependence in binary panel data with individual covariates. For this aim, we rely on a quadratic exponential model in which the association between the response variables is accounted for in a different way with respect to more standard formulations. The level of association is measured by a single parameter that may be estimated by a Conditional Maximum Likelihood (CML) approach. Under the dynamic logit model, the conditional estimator of this parameter converges to zero when the hypothesis of absence of state dependence is true. Therefore, it is possible to implement a t-test for this hypothesis which may be very simply performed and attains the nominal significance level under several structures of the individual covariates. Through an extensive simulation study, we find that our test has good finite sample properties and it is more robust to the presence of (autocorrelated) covariates in the model specification in comparison with other existing testing procedures for state dependence. The proposed approach is illustrated by two empirical applications: the first is based on data coming from the Panel Study of Income Dynamics and concerns employment and fertility; the second is based on the Health and Retirement Study and concerns the self reported health status.     

Action following the discovery of a global association between the whole genome and adverse event risk in a clinical drug‐development programme

Observation of adverse drug reactions during drug development can cause closure of the whole programme. However, if association between the genotype and the risk of an adverse event is discovered, then it might suffice to exclude patients of certain genotypes from future recruitment. Various sequential and non‐sequential procedures are available to identify an association between the whole genome, or at least a portion of it, and the incidence of adverse events. In this paper we start with a suspected association between the genotype and the risk of an adverse event and suppose that the genetic subgroups with elevated risk can be identified. Our focus is determination of whether the patients identified as being at risk should be excluded from further studies of the drug. We propose using a utility function to determine the appropriate action, taking into account the relative costs of suffering an adverse reaction and of failing to alleviate the patient's disease. Two illustrative examples are presented, one comparing patients who suffer from an adverse event with contemporary patients who do not, and the other making use of a reference control group. We also illustrate two classification methods, LASSO and CART, for identifying patients at risk, but we stress that any appropriate classification method could be used in conjunction with the proposed utility function. Our emphasis is on determining the action to take rather than on providing definitive evidence of an association. Copyright © 2008 John Wiley & Sons, Ltd.