In recent years, advances in high throughput sequencing technology have led to a need for specialized methods for the analysis of digital gene expression data. While gene expression data measured on a microarray take on continuous values and can be modeled using the normal distribution, RNA sequencing data involve nonnegative counts and are more appropriately modeled using a discrete count distribution, such as the Poisson or the negative binomial. Consequently, analytic tools that assume a Gaussian distribution (such as classification methods based on linear discriminant analysis and clustering methods that use Euclidean distance) may not perform as well for sequencing data as methods that are based upon a more appropriate distribution. Here, we propose new approaches for performing classification and clustering of observations on the basis of sequencing data. Using a Poisson log linear model, we develop an analog of diagonal linear discriminant analysis that is appropriate for sequencing data. We also propose an approach for clustering sequencing data using a new dissimilarity measure that is based upon the Poisson model. We demonstrate the performances of these approaches in a simulation study, on three publicly available RNA sequencing data sets, and on a publicly available chromatin immunoprecipitation sequencing data set.
We propose a flexible change-point model for inhomogeneous Poisson Processes, which arise naturally from next-generation DNA sequencing, and derive score and generalized likelihood statistics for shifts in intensity functions. We construct a modified Bayesian information criterion (mBIC) to guide model selection, and point-wise approximate Bayesian confidence intervals for assessing the confidence in the segmentation. The model is applied to DNA Copy Number profiling with sequencing data and evaluated on simulated spike-in and real data sets.
Gaussian graphical models are often used to infer gene networks based on microarray expression data. Many scientists, however, have begun using high-throughput sequencing technologies to measure gene expression. As the resulting high-dimensional count data consists of counts of sequencing reads for each gene, Gaussian graphical models are not optimal for modeling gene networks based on this discrete data. We develop a novel method for estimating high-dimensional Poisson graphical models, the Log-Linear Graphical Model, allowing us to infer networks based on high-throughput sequencing data. Our…
We introduce the Poisson Log-Normal Graphical Model for count data, and present a normality transformation for data arising from this distribution. The model and transformation are feasible for high-throughput microRNA (miRNA) sequencing data and directly account for known overdispersion relationships present in this data set. The model allows for network dependencies to be modeled, and we provide an algorithm which utilizes a one-step EM based result in order to allow for a provable increase in performance in determining the network structure. The model is shown to provide an increase in perf…
Identification of clusters of co-expressed genes in transcriptomic data is a difficult task. Most algorithms used for this purpose can be classified into two broad categories: distance-based or model-based approaches. Distance-based approaches typically utilize a distance function between pairs of data objects and group similar objects together into clusters. Model-based approaches are based on using the mixture-modeling framework. Compared to distance-based approaches, model-based approaches offer better interpretability because each cluster can be explicitly characterized in terms of the pro…
The detection of local genomic signals using high-throughput DNA sequencing data can be cast as a problem of scanning a Poisson random field for local changes in the rate of the process. We propose a likelihood-based framework for for such scans, and derive formulas for false positive rate control and power calculations. The framework can also accommodate mixtures of Poisson processes to deal with over-dispersion. As a specific, detailed example, we consider the detection of insertions and deletions by paired-end DNA-sequencing. We propose several statistics for this problem, compare their pow…
With ongoing developments and innovations in single-cell RNA sequencing methods, advancements in sequencing performance could empower significant discoveries as well as new emerging possibilities to address biological and medical investigations. In the study, we will be using the dataset collected by the authors of Systematic comparative analysis of single cell RNA-sequencing methods. The dataset consists of single-cell and single nucleus profiling from three types of samples - cell lines, peripheral blood mononuclear cells, and brain tissue, which offers 36 libraries in six separate experimen…
Background: We study the statistical properties of fragment coverage in genome sequencing experiments. In an extension of the classic Lander-Waterman model, we consider the effect of the length distribution of fragments. We also introduce the notion of the shape of a coverage function, which can be used to detect abberations in coverage. The probability theory underlying these problems is essential for constructing models of current high-throughput sequencing experiments, where both sample preparation protocols and sequencing technology particulars can affect fragment length distributions. Res…
Pathogen deep-sequencing is an increasingly routinely used technology in infectious disease surveillance. We present a semi-parametric Bayesian Poisson model to exploit these emerging data for inferring infectious disease transmission flows and the sources of infection at the population level. The framework is computationally scalable in high dimensional flow spaces thanks to Hilbert Space Gaussian process approximations, allows for sampling bias adjustments, and estimation of gender- and age-specific transmission flows at finer resolution than previously possible. We apply the approach to den…
In our Brief Communication (DOI: 10.1038/s41592-018-0033-z), we presented the method SAVER for recovering true gene expression levels in noisy single cell RNA sequencing data. We evaluated the performance of SAVER, along with comparable methods MAGIC and scImpute, in an RNA FISH validation experiment and a data downsampling experiment. In a Comment [arXiv:1908.07084v1], Li & Li were concerned with the use of the downsampled datasets, specifically focusing on clustering results obtained from the Zeisel et al. data. Here, we will address these comments and, furthermore, amend the data downsampli…
In the analysis of single-cell RNA sequencing data, researchers often characterize the variation between cells by estimating a latent variable, such as cell type or pseudotime, representing some aspect of the individual cell's state. They then test each gene for association with the estimated latent variable. If the same data are used for both of these steps, then standard methods for computing p-values and confidence intervals in the second step will fail to achieve statistical guarantees such as Type 1 error control. Furthermore, approaches such as sample splitting that can be applied to sol…
We present a new modelling approach for longitudinal count data that is motivated by the increasing availability of longitudinal RNA-sequencing experiments. The distribution of RNA-seq counts typically exhibits overdispersion, zero-inflation and heavy tails; moreover, in longitudinal designs repeated measurements from the same subject are typically (positively) correlated. We propose a generalized linear mixed model based on the Poisson-Tweedie distribution that can flexibly handle each of the aforementioned features of longitudinal overdispersed counts. We develop a computational approach to …
On June 25th, 2018, Huang et al. published a computational method SAVER on Nature Methods for imputing dropout gene expression levels in single cell RNA sequencing (scRNA-seq) data. Huang et al. performed a set of comprehensive benchmarking analyses, including comparison with the data from RNA fluorescence in situ hybridization, to demonstrate that SAVER outperformed two existing scRNA-seq imputation methods, scImpute and MAGIC. However, their computational analyses were based on semi-synthetic data that the authors had generated following the Poisson-Gamma model used in the SAVER method. We h…
Cancer is a number of related yet highly heterogeneous diseases. Correct identification of cancer subtypes is critical for clinical decisions. The advance in sequencing technologies has made it possible to study cancer based on abundant genomics and transcriptomic (-omics) data. Such a data-driven approach is expected to address limitations and issues with traditional methods in identifying cancer subtypes. We evaluate the suitability of clustering--a data mining tool to study heterogenous data when there is a lack of sufficient understanding of the subject matters--in the identification of ca…
Model-based clustering is a technique widely used to group a collection of units into mutually exclusive groups. There are, however, situations in which an observation could in principle belong to more than one cluster. In the context of Next-Generation Sequencing (NGS) experiments, for example, the signal observed in the data might be produced by two (or more) different biological processes operating together and a gene could participate in both (or all) of them. We propose a novel approach to cluster NGS discrete data, coming from a ChIP-Seq experiment, with a mixture model, allowing each un…
RNA-sequencing (RNA-Seq) has become a powerful technology to characterize gene expression profiles because it is more accurate and comprehensive than microarrays. Although statistical methods that have been developed for microarray data can be applied to RNA-Seq data, they are not ideal due to the discrete nature of RNA-Seq data. The Poisson distribution and negative binomial distribution are commonly used to model count data. Recently, Witten (2011) proposed a Poisson linear discriminant analysis for RNA-Seq data. The Poisson assumption may not be as appropriate as negative binomial distribut…
We study the influence of clustering, more specifically triangles, on cascading failures in interdependent networks or systems, in which we model the dependence between comprising systems using a dependence graph. First, we propose a new model that captures how the presence of triangles in the dependence graph alters the manner in which failures transmit from affected systems to others. Unlike existing models, the new model allows us to approximate the failure propagation dynamics using a multi-type branching process, even with triangles. Second, making use of the model, we provide a simple co…
The advent of single-cell sequencing opens new avenues for personalized treatment. In this paper, we address a two-level clustering problem of simultaneous subject subgroup discovery (subject level) and cell type detection (cell level) for single-cell expression data from multiple subjects. However, current statistical approaches either cluster cells without considering the subject heterogeneity or group subjects without using the single-cell information. To bridge the gap between cell clustering and subject grouping, we develop a nonparametric Bayesian model, Subject and Cell clustering for S…
Spatial transcriptomics is a modern sequencing technology that allows the measurement of the activity of thousands of genes in a tissue sample and map where the activity is occurring. This technology has enabled the study of the so-called spatially expressed genes, i.e., genes which exhibit spatial variation across the tissue. Comprehending their functions and their interactions in different areas of the tissue is of great scientific interest, as it might lead to a deeper understanding of several key biological mechanisms. However, adequate statistical tools that exploit the newly spatial mapp…
Viral deep-sequencing data play a crucial role toward understanding disease transmission network flows, because the higher resolution of these data compared to standard Sanger sequencing provide evidence into the direction of infectious disease transmission. To more fully utilize these rich data and account for the uncertainties in phylogenetic analysis outcomes, we propose a spatial Poisson process model to uncover HIV transmission flow patterns at the population level. We represent pairings of two individuals with viral sequence data as typed points, with coordinates representing covariates …
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…
Robust Bayesian Tensor Factorization with Zero-Inflated Poisson Model and Consensus Aggregation
Broad Institute of MIT and Harvard（麻省理工学院与哈佛大学博德研究所）；Klarman Cell Observatory, Broad Institute of MIT and Harvard（麻省理工学院与哈佛大学博德研究所克拉曼细胞观测站）；Brigham and Women’s Hospital（布莱根妇女医院）；Harvard Medical School（哈佛医学院）；University of Massachusetts Boston（马萨诸塞大学波士顿分校）
