Gene-based interaction analysis shows GABAergic genes interacting with parenting in adolescent depressive symptoms.

BACKGROUND: Most gene-environment interaction studies (G × E) have focused on single candidate genes. This approach is criticized for its expectations of large effect sizes and occurrence of spurious results. We describe an approach that accounts for the polygenic nature of most psychiatric phenotypes and reduces the risk of false-positive findings. We apply this method focusing on the role of perceived parental support, psychological control, and harsh punishment in depressive symptoms in adolescence. METHODS: Analyses were conducted on 982 adolescents of Caucasian origin (Mage (SD) = 13.78 (.94) years) genotyped for 4,947 SNPs in 263 genes, selected based on a literature survey. The Leuven Adolescent Perceived Parenting Scale (LAPPS) and the Parental Behavior Scale (PBS) were used to assess perceived parental psychological control, harsh punishment, and support. The Center for Epidemiologic Studies Depression Scale (CES-D) was the outcome. We used gene-based testing taking into account linkage disequilibrium to identify genes containing SNPs exhibiting an interaction with environmental factors yielding a p-value per single gene. Significant results at the corrected p-value of p < 1.90 × 10-4 were examined in an independent replication sample of Dutch adolescents (N = 1354). RESULTS: Two genes showed evidence for interaction with perceived support: GABRR1 (p = 4.62 × 10-5 ) and GABRR2 (p = 9.05 × 10-6 ). No genes interacted significantly with psychological control or harsh punishment. Gene-based analysis was unable to confirm the interaction of GABRR1 or GABRR2 with support in the replication sample. However, for GABRR2, but not GABRR1, the correlation of the estimates between the two datasets was significant (r (46) = .32; p = .027) and a gene-based analysis of the combined datasets supported GABRR2 × support interaction (p = 1.63 × 10-4 ). CONCLUSIONS: We present a gene-based method for gene-environment interactions in a polygenic context and show that genes interact differently with particular aspects of parenting. This accentuates the importance of polygenic approaches and the need to accurately assess environmental exposure in G × E.

A Comparison of Methods for Gene-Based Testing That Account for Linkage Disequilibrium.

Controlling the type I error rate while retaining sufficient power is a major concern in genome-wide association studies, which nowadays often examine more than a million single-nucleotide polymorphisms (SNPs) simultaneously. Methods such as the Bonferroni correction can lead to a considerable decrease in power due to the large number of tests conducted. Shifting the focus to higher functional structures (e.g., genes) can reduce the loss of power. This can be accomplished via the combination of p-values of SNPs that belong to the same structural unit to test their joint null hypothesis. However, standard methods for this purpose (e.g., Fisher's method) do not account for the dependence among the tests due to linkage disequilibrium (LD). In this paper, we review various adjustments to methods for combining p-values that take LD information explicitly into consideration and evaluate their performance in a simulation study based on data from the HapMap project. The results illustrate the importance of incorporating LD information into the methods for controlling the type I error rate at the desired level. Furthermore, some methods are more successful in controlling the type I error rate than others. Among them, Brown's method was the most robust technique with respect to the characteristics of the genes and outperformed the Bonferroni method in terms of power in many scenarios. Examining the genetic factors of a phenotype of interest at the gene-rather than SNP-level can provide researchers benefits in terms of the power of the study. While doing so, one should be careful to account for LD in SNPs belonging to the same gene, for which Brown's method seems the most robust technique.

Testing Gene-Gene Interactions Based on a Neighborhood Perspective in Genome-wide Association Studies.

Unexplained genetic variation that causes complex diseases is often induced by gene-gene interactions (GGIs). Gene-based methods are one of the current statistical methodologies for discovering GGIs in case-control genome-wide association studies that are not only powerful statistically, but also interpretable biologically. However, most approaches include assumptions about the form of GGIs, which results in poor statistical performance. As a result, we propose gene-based testing based on the maximal neighborhood coefficient (MNC) called gene-based gene-gene interaction through a maximal neighborhood coefficient (GBMNC). MNC is a metric for capturing a wide range of relationships between two random vectors with arbitrary, but not necessarily equal, dimensions. We established a statistic that leverages the difference in MNC in case and in control samples as an indication of the existence of GGIs, based on the assumption that the joint distribution of two genes in cases and controls should not be substantially different if there is no interaction between them. We then used a permutation-based statistical test to evaluate this statistic and calculate a statistical p-value to represent the significance of the interaction. Experimental results using both simulation and real data showed that our approach outperformed earlier methods for detecting GGIs.

Gene-Based Testing of Interactions Using XGBoost in Genome-Wide Association Studies.

Among the myriad of statistical methods that identify gene-gene interactions in the realm of qualitative genome-wide association studies, gene-based interactions are not only powerful statistically, but also they are interpretable biologically. However, they have limited statistical detection by making assumptions on the association between traits and single nucleotide polymorphisms. Thus, a gene-based method (GGInt-XGBoost) originated from XGBoost is proposed in this article. Assuming that log odds ratio of disease traits satisfies the additive relationship if the pair of genes had no interactions, the difference in error between the XGBoost model with and without additive constraint could indicate gene-gene interaction; we then used a permutation-based statistical test to assess this difference and to provide a statistical p-value to represent the significance of the interaction. Experimental results on both simulation and real data showed that our approach had superior performance than previous experiments to detect gene-gene interactions.

Integration of Enhancer-Promoter Interactions with GWAS Summary Results Identifies Novel Schizophrenia-Associated Genes and Pathways.

It remains challenging to boost statistical power of genome-wide association studies (GWASs) to identify more risk variants or loci that can account for "missing heritability." Furthermore, since most identified variants are not in gene-coding regions, a biological interpretation of their function is largely lacking. On the other hand, recent biotechnological advances have made it feasible to experimentally measure the three-dimensional organization of the genome, including enhancer-promoter interactions in high resolutions. Due to the well-known critical roles of enhancer-promoter interactions in regulating gene expression programs, such data have been applied to link GWAS risk variants to their putative target genes, gaining insights into underlying biological mechanisms. However, their direct use in GWAS association testing is yet to be exploited. Here we propose integrating enhancer-promoter interactions into GWAS association analysis to both boost statistical power and enhance interpretability. We demonstrate that through an application to two large-scale schizophrenia (SCZ) GWAS summary data sets, the proposed method could identify some novel SCZ-associated genes and pathways (containing no significant SNPs). For example, after the Bonferroni correction, for the larger SCZ data set with 36,989 cases and 113,075 controls, our method applied to the gene body and enhancer regions identified 27 novel genes and 11 novel KEGG pathways to be significant, all missed by the transcriptome-wide association study (TWAS) approach. We conclude that our proposed method is potentially useful and is complementary to TWAS and other standard gene- and pathway-based methods.

Improved Use of Small Reference Panels for Conditional and Joint Analysis with GWAS Summary Statistics.

Due to issues of practicality and confidentiality of genomic data sharing on a large scale, typically only meta- or mega-analyzed genome-wide association study (GWAS) summary data, not individual-level data, are publicly available. Reanalyses of such GWAS summary data for a wide range of applications have become more and more common and useful, which often require the use of an external reference panel with individual-level genotypic data to infer linkage disequilibrium (LD) among genetic variants. However, with a small sample size in only hundreds, as for the most popular 1000 Genomes Project European sample, estimation errors for LD are not negligible, leading to often dramatically increased numbers of false positives in subsequent analyses of GWAS summary data. To alleviate the problem in the context of association testing for a group of SNPs, we propose an alternative estimator of the covariance matrix with an idea similar to multiple imputation. We use numerical examples based on both simulated and real data to demonstrate the severe problem with the use of the 1000 Genomes Project reference panels, and the improved performance of our new approach.