Microbes can affect processes from food production to human health. Such microbes are not isolated, but rather interact with each other and establish connections with their living environments. Understanding these interactions is essential to an understanding of the organization and complex interplay of microbial communities, as well as the structure and dynamics of various ecosystems. A common and essential approach toward this objective involves the inference of microbiome interaction networks. Although network inference methods in other fields have been studied before, applying these methods to estimate microbiome associations based on compositional data will not yield valid results. On the one hand, features of microbiome data such as compositionality, sparsity and high-dimensionality challenge the data normalization and the design of computational methods. On the other hand, several issues like microbial community heterogeneity, external environmental interference and biological concerns also make it more difficult to deal with the network inference. In this paper, we provide a comprehensive review of emerging microbiome interaction network inference methods. According to various assumptions and research targets, estimated networks are divided into four main categories: correlation networks, conditional correlation networks, mixture networks and differential networks. Their scope of applications, advantages and limitations are presented in this review. Since real microbial interactions can be complex and dynamic, no unifying method has captured all the aspects of interest to date. In addition, we discuss the challenges now confronting current microbial associations study and future prospects. Finally, we highlight that the research in microbial network inference requires the joint promotion of statistical computation methods and experimental techniques.
One important problem in microbiome analysis is to identify the bacterial taxa that are associated with a response, where the microbiome data are summarized as the composition of the bacterial taxa at different taxonomic levels. This paper considers regression analysis with such compositional data as covariates. In order to satisfy the subcompositional coherence of the results, linear models with a set of linear constraints on the regression coefficients are introduced. Such models allow regression analysis for subcompositions and include the log-contrast model for compositional covariates as …
The human microbiome plays critical roles in human health and has been linked to many diseases. While advanced sequencing technologies can characterize the composition of the microbiome in unprecedented detail, it remains challenging to disentangle the complex interplay between human microbiome and disease risk factors due to the complicated nature of microbiome data. Excessive numbers of zero values, high dimensionality, the hierarchical phylogenetic tree and compositional structure are compounded and consequently make existing methods inadequate to appropriately address these issues. We prop…
Microbiome data are complex in nature, involving high dimensionality, compositionally, zero inflation, and taxonomic hierarchy. Compositional data reside in a simplex that does not admit the standard Euclidean geometry. Most existing compositional regression methods rely on transformations that are inadequate or even inappropriate in modeling data with excessive zeros and taxonomic structure. We develop a novel relative-shift regression framework that directly uses compositions as predictors. The new framework provides a paradigm shift for compositional regression and offers a superior biologi…
The neonatal intensive care unit (NICU) experience is known to be one of the most crucial factors that drive preterm infant's neurodevelopmental and health outcomes. It is hypothesized that stressful early life experience of very preterm neonate is imprinting gut microbiome by the regulation of the so-called brain-gut axis, and consequently, certain microbiome markers are predictive of later infant neurodevelopment. To investigate, a preterm infant study was conducted; infant fecal samples were collected during the infants' first month of postnatal age, resulting in functional compositional mi…
The so-called gut-brain axis has stimulated extensive research on microbiomes. One focus is to assess the association between certain clinical outcomes and the relative abundances of gut microbes, which can be presented as sub-compositional data in conformity with the taxonomic hierarchy of bacteria. Motivated by a study for identifying the microbes in the gut microbiome of preterm infants that impact their later neurobehavioral outcomes, we formulate a constrained integrative multi-view regression, where the neurobehavioral scores form multivariate response, the sub-compositional microbiome d…
This paper is motivated by the recent interest in the analysis of high dimen- sional microbiome data. A key feature of this data is the presence of `structural zeros' which are microbes missing from an observation vector due to an underlying biological process and not due to error in measurement. Typical notions of missingness are insufficient to model these structural zeros. We define a general framework which allows for structural zeros in the model and propose methods of estimating sparse high dimensional covariance and precision matrices under this setup. We establish error bounds in the s…
Bayesian Sparse Regression for Microbiome-Metabolite Data Integration
The University of Texas Health Science Center at Houston（德克萨斯大学健康科学中心休斯顿分校）；The University of Texas MD Anderson Cancer Center（德克萨斯大学安德森癌症中心）；Rice University（莱斯大学）
Testing Microbiome Community Differences in High Dimensions: A Bootstrap Approach for Compositional Data
Indian Institute of Technology Bombay（印度理工学院孟买分校）；Missouri University of Science and Technology（密苏里科技大学）；Washington University in St. Louis（圣路易斯华盛顿大学）；University of Missouri（密苏里大学）；University of Michigan（密歇根大学）
Identifying which taxa in our microbiota are associated with traits of interest is important for advancing science and health. However, the identification is challenging because the measured vector of taxa counts (by amplicon sequencing) is compositional, so a change in the abundance of one taxon in the microbiota induces a change in the number of sequenced counts across all taxa. The data is typically sparse, with zero counts present either due to biological variance or limited sequencing depth (technical zeros). For low abundance taxa, the chance for technical zeros is non-negligible. We sho…
The human microbiome has an important role in determining health. Mediation analyses quantify the contribution of the microbiome in the causal path between exposure and disease; however, current mediation models cannot fully capture the high dimensional, correlated, and compositional nature of microbiome data and do not typically accommodate dichotomous outcomes. We propose a novel approach that uses inverse odds weighting to test for the mediating effect of the microbiome. We use simulation to demonstrate that our approach gains power for high dimensional mediators, and it is agnostic to the …
An important task in microbiome studies is to test the existence of and give characterization to differences in the microbiome composition across groups of samples. Important challenges of this problem include the large within-group heterogeneities among samples and the existence of potential confounding variables that, when ignored, increase the chance of false discoveries and reduce the power for identifying true differences. We propose a probabilistic framework to overcome these issues by combining three ideas: (i) a phylogenetic tree-based decomposition of the cross-group comparison proble…
Recent attacks of various viruses with having deep and extensive impact at a global scale has warranted that microbiome be studied extensively and in a robust analytic framework. Microbiome typically refers to the collective genomes of such organisms, although it could also refer to the collection of the organisms by themselves. Here we provide an overview of statistical techniques that are useful in analysing such data.
One of the major research questions regarding human microbiome studies is the feasibility of designing interventions that modulate the composition of the microbiome to promote health and cure disease. This requires extensive understanding of the modulating factors of the microbiome, such as dietary intake, as well as the relation between microbial composition and phenotypic outcomes, such as body mass index (BMI). Previous efforts have modeled these data separately, employing two-step approaches that can produce biased interpretations of the results. Here, we propose a Bayesian joint model tha…
High-throughput sequencing technology allows us to test the compositional difference of bacteria in different populations. One important feature of human microbiome data is that it often includes a large number of zeros. Such data can be treated as being generated from a two-part model that includes a zero point-mass. Motivated by analysis of such non-negative data with excessive zeros, we introduce several truncated rank-based two-group and multi-group tests for such data, including a truncated rank-based Wilcoxon rank-sum test for two-group comparison and two truncated Kruskal-Wallis tests f…
Instrumental Variable Estimation for Compositional Treatments
Helmholtz Munich（亥姆霍兹慕尼黑中心）；TUM School of Computation, Information and Technology, Technical University of Munich（慕尼黑工业大学计算、信息与技术学院）；Munich Center for Machine Learning (MCML)（慕尼黑机器学习中心）；Ludwig-Maximilian University（路德维希-马克西米利安大学）；Center for Computational Mathematics, Flatiron Institute（弗拉蒂伦研究所计算数学中心）
The analysis of human microbiome data is often based on dimension-reduced graphical displays and clustering derived from vectors of microbial abundances in each sample. Common to these ordination methods is the use of biologically motivated definitions of similarity. Principal coordinate analysis, in particular, is often performed using ecologically defined distances, allowing analyses to incorporate context-dependent, non-Euclidean structure. Here we describe how to take a step beyond ordination plots and incorporate this structure into high-dimensional penalized regression models. Within thi…
Comparison of statistical methods for high-dimensional compositional data from flow cytometry: A critical perspective on log-ratio transformation and differential abundance testing
National Cancer Institute/National Institutes of Health（美国国立癌症研究所/美国国立卫生研究院）
Quantifying the relation between gut microbiome and body weight can provide insights into personalized strategies for improving digestive health. In this paper, we present an algorithm that predicts weight fluctuations using gut microbiome in a healthy cohort of newborns from a previously published dataset. Microbial data has been known to present unique statistical challenges that defy most conventional models. We propose a mixed effect Dirichlet-tree multinomial (DTM) model to untangle these difficulties as well as incorporate covariate information and account for species relatedness. The DT…
A network lasso enables us to construct a model for each sample, which is known as multi-task learning. Existing methods for multi-task learning cannot be applied to compositional data due to their intrinsic properties. In this paper, we propose a multi-task learning method for compositional data using a sparse network lasso. We focus on a symmetric form of the log-contrast model, which is a regression model with compositional covariates. The effectiveness of the proposed method is shown through simulation studies and application to gut microbiome data.
The target of inference in microbiome analyses is usually relative abundance (RA) because RA in a sample (e.g., stool) can be considered as an approximation of RA in an entire ecosystem (e.g., gut). However, inference on RA suffers from the fact that RA are calculated by dividing absolute abundances (AA) over the common denominator (CD), the summation of all AA (i.e., library size). Because of that, perturbation in one taxon will result in a change in the CD and thus cause false changes in RA of all other taxa, and those false changes could lead to false positive/negative findings. We propose …
Studying the human microbiome has gained substantial interest in recent years, and a common task in the analysis of these data is to cluster microbiome compositions into subtypes. This subdivision of samples into subgroups serves as an intermediary step in achieving personalized diagnosis and treatment. In applying existing clustering methods to modern microbiome studies including the American Gut Project (AGP) data, we found that this seemingly standard task, however, is very challenging in the microbiome composition context due to several key features of such data. Standard distance-based cl…
Mixed-membership (MM) models such as Latent Dirichlet Allocation (LDA) have been applied to microbiome compositional data to identify latent subcommunities of microbial species. These subcommunities are informative for understanding the biological interplay of microbes and for predicting health outcomes. However, microbiome compositions typically display substantial cross-sample heterogeneities in subcommunity compositions -- that is, the variability in the proportions of microbes in shared subcommunities across samples -- which is not accounted for in prior analyses. As a result, LDA can prod…
There is a growing awareness of the important roles that microbial communities play in complex biological processes. Modern investigation of these often uses next generation sequencing of metagenomic samples to determine community composition. We propose a statistical technique based on clique loglinear models and Bayes model averaging to identify microbial components in a metagenomic sample at various taxonomic levels that have significant associations. We describe the model class, a stochastic search technique for model selection, and the calculation of estimates of posterior probabilities o…
The study of immune cellular composition has been of great scientific interest in immunology because of the generation of multiple large-scale data. From the statistical point of view, such immune cellular data should be treated as compositional. In compositional data, each element is positive, and all the elements sum to a constant, which can be set to one in general. Standard statistical methods are not directly applicable for the analysis of compositional data because they do not appropriately handle correlations between the compositional elements. In this paper, we review statistical metho…
Differential abundance tests in compositional data are essential and fundamental tasks in various biomedical applications, such as single-cell, bulk RNA-seq, and microbiome data analysis. However, because of the compositional constraint and the prevalence of zero counts in the data, differential abundance analysis in compositional data remains a complicated and unsolved statistical problem. This study introduces a new differential abundance test, the robust differential abundance (RDB) test, to address these challenges. Compared with existing methods, the RDB test is simple and computationally…
Applications such as the analysis of microbiome data have led to renewed interest in statistical methods for compositional data, i.e., multivariate data in the form of probability vectors that contain relative proportions. In particular, there is considerable interest in modeling interactions among such relative proportions. To this end we propose a class of exponential family models that accommodate general patterns of pairwise interaction while being supported on the probability simplex. Special cases include the family of Dirichlet distributions as well as Aitchison's additive logistic norm…
Many biological high-throughput data sets, such as targeted amplicon-based and metagenomic sequencing data, are compositional in nature. A common exploratory data analysis task is to infer statistical associations between the high-dimensional microbial compositions and habitat- or host-related covariates. We propose a general robust statistical regression framework, RobRegCC (Robust Regression with Compositional Covariates), which extends the linear log-contrast model by a mean shift formulation for capturing outliers. RobRegCC includes sparsity-promoting convex and non-convex penalties for pa…
In microbiome and genomic studies, the regression of compositional data has been a crucial tool for identifying microbial taxa or genes that are associated with clinical phenotypes. To account for the variation in sequencing depth, the classic log-contrast model is often used where read counts are normalized into compositions. However, zero read counts and the randomness in covariates remain critical issues. In this article, we introduce a surprisingly simple, interpretable, and efficient method for the estimation of compositional data regression through the lens of a novel high-dimensional lo…
In recent years microbiome studies have become increasingly prevalent and large-scale. Through high-throughput sequencing technologies and well-established analytical pipelines, relative abundance data of operational taxonomic units and their associated taxonomic structures are routinely produced. Since such data can be extremely sparse and high dimensional, there is often a genuine need for dimension reduction to facilitate data visualization and downstream statistical analysis. We propose Principal Amalgamation Analysis (PAA), a novel amalgamation-based and taxonomy-guided dimension reductio…
Microorganisms play critical roles in human health and disease. It is well known that microbes live in diverse communities in which they interact synergistically or antagonistically. Thus for estimating microbial associations with clinical covariates, multivariate statistical models are preferred. Multivariate models allow one to estimate and exploit complex interdependencies among multiple taxa, yielding more powerful tests of exposure or treatment effects than application of taxon-specific univariate analyses. In addition, the analysis of microbial count data requires special attention becau…
Microbiota contribute to many dimensions of host phenotype, including disease. To link specific microbes to specific phenotypes, microbiome-wide association studies compare microbial abundances between two groups of samples. Abundance differences, however, reflect not only direct associations with the phenotype, but also indirect effects due to microbial interactions. We found that microbial interactions could easily generate a large number of spurious associations that provide no mechanistic insight. Using techniques from statistical physics, we developed a method to remove indirect associati…
Microbial ecology serves as a foundation for a wide range of scientific and biomedical studies. Rapidly-evolving high-throughput sequencing technology enables the comprehensive search for microbial biomarkers using longitudinal experiments. Such experiments consist of repeated biological observations from each subject over time and are essential in accounting for the high between-subject and within-subject variability. Unfortunately, many of the statistical tests based on parametric models rely on correctly specifying temporal dependence structure which is unavailable in most microbiome data. …
The interactions between microbial taxa in microbiome data has been under great research interest in the science community. In particular, several methods such as SPIEC-EASI, gCoda, and CD-trace have been proposed to model the conditional dependency between microbial taxa, in order to eliminate the detection of spurious correlations. However, all those methods are built upon the central log-ratio (CLR) transformation, which results in a degenerate covariance matrix and thus an undefined inverse covariance matrix as the estimation of the underlying network. Jiang et al. (2021) and Tian et al. (…
The Dirichlet-multinomial (DM) distribution plays a fundamental role in modern statistical methodology development and application. Recently, the DM distribution and its variants have been used extensively to model multivariate count data generated by high-throughput sequencing technology in omics research due to its ability to accommodate the compositional structure of the data as well as overdispersion. A major limitation of the DM distribution is that it is unable to handle excess zeros typically found in practice which may bias inference. To fill this gap, we propose a novel Bayesian zero-…
Inverse Probability Weighting-based Mediation Analysis for Microbiome Data
The University of Texas at San Antonio（德克萨斯大学圣安东尼奥分校）；The University of Texas MD Anderson Cancer Center（德克萨斯大学安德森癌症中心）；BeOne Medicines（博生医药）；Texas A&M University（德克萨斯农工大学）；University of Toronto（多伦多大学）；Columbia University（哥伦比亚大学）
Dissecting Microbial Community Structure and Heterogeneity via Multivariate Covariate-Adjusted Clustering
University of Connecticut Health Center（康涅狄格大学健康中心）；Department of Biostatistics, University of Michigan（密歇根大学生物统计系）；Department of Statistics, University of Connecticut（康涅狄格大学统计学系）
Understanding how microbes interact with each other is key to revealing the underlying role that microorganisms play in the host or environment and to identifying microorganisms as an agent that can potentially alter the host or environment. For example, understanding how the microbial interactions associate with parasitic infection can help resolve potential drug or diagnostic test for parasitic infection. To unravel the microbial interactions, existing tools often rely on graphical models to infer the conditional dependence of microbial abundances to represent their interactions. However, cu…
