scholarly journals glmGamPoi: Fitting Gamma-Poisson Generalized Linear Models on Single Cell Count Data

Author(s):  
Constantin Ahlmann-Eltze ◽  
Wolfgang Huber

Abstract Motivation The Gamma-Poisson distribution is a theoretically and empirically motivated model for the sampling variability of single cell RNA-sequencing counts (Grün et al., 2014; Svensson, 2020; Silverman et al., 2018; Hafemeister and Satija, 2019) and an essential building block for analysis approaches including differential expression analysis (Robinson et al., 2010; McCarthy et al., 2012; Anders and Huber, 2010; Love et al., 2014), principal component analysis (Townes et al., 2019) and factor analysis (Risso et al., 2018). Existing implementations for inferring its parameters from data often struggle with the size of single cell datasets, which can comprise millions of cells; at the same time, they do not take full advantage of the fact that zero and other small numbers are frequent in the data. These limitations have hampered uptake of the model, leaving room for statistically inferior approaches such as logarithm(-like) transformation. Results We present a new R package for fitting the Gamma-Poisson distribution to data with the characteristics of modern single cell datasets more quickly and more accurately than existing methods. The software can work with data on disk without having to load them into RAM simultaneously. Availability The package glmGamPoi is available from Bioconductor for Windows, macOS, and Linux, and source code is available on github.com/const-ae/glmGamPoi under a GPL-3 license.

Author(s):  
Constantin Ahlmann-Eltze ◽  
Wolfgang Huber

AbstractMotivationThe Gamma-Poisson distribution is a theoretically and empirically motivated model for the sampling variability of single cell RNA-sequencing counts (Grün et al., 2014; Townes et al., 2019; Svensson, 2020; Silverman et al., 2018; Hafemeister and Satija, 2019) and an essential building block for analysis approaches including differential expression analysis (Robinson et al., 2010; McCarthy et al., 2012; Anders and Huber, 2010; Love et al., 2014), principal component analysis (Townes et al., 2019) and factor analysis (Risso et al., 2018). Existing implementations for inferring its parameters from data often struggle with the size of single cell datasets, which typically comprise thousands or millions of cells; at the same time, they do not take full advantage of the fact that zero and other small numbers are frequent in the data. These limitations have hampered uptake of the model, leaving room for statistically inferior approaches such as logarithm(-like) transformation.ResultsWe present a new R package for fitting the Gamma-Poisson distribution to data with the characteristics of modern single cell datasets more quickly and more accurately than existing methods. The software can work with data on disk without having to load them into RAM simultaneously.AvailabilityThe package glmGamPoi is available from Bioconductor (since release 3.11) for Windows, macOS, and Linux, and source code is available on GitHub under a GPL-3 license. The scripts to reproduce the results of this paper are available on GitHub as [email protected]


PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e10469
Author(s):  
Daniel Dimitrov ◽  
Quan Gu

Background RNA sequencing is an indispensable research tool used in a broad range of transcriptome analysis studies. The most common application of RNA Sequencing is differential expression analysis and it is used to determine genetic loci with distinct expression across different conditions. An emerging field called single-cell RNA sequencing is used for transcriptome profiling at the individual cell level. The standard protocols for both of these approaches include the processing of sequencing libraries and result in the generation of count matrices. An obstacle to these analyses and the acquisition of meaningful results is that they require programing expertise. Although some effort has been directed toward the development of user-friendly RNA-Seq analysis analysis tools, few have the flexibility to explore both Bulk and single-cell RNA sequencing. Implementation BingleSeq was developed as an intuitive application that provides a user-friendly solution for the analysis of count matrices produced by both Bulk and Single-cell RNA-Seq experiments. This was achieved by building an interactive dashboard-like user interface which incorporates three state-of-the-art software packages for each type of the aforementioned analyses. Furthermore, BingleSeq includes additional features such as visualization techniques, extensive functional annotation analysis and rank-based consensus for differential gene analysis results. As a result, BingleSeq puts some of the best reviewed and most widely used packages and tools for RNA-Seq analyses at the fingertips of biologists with no programing experience. Availability BingleSeq is as an easy-to-install R package available on GitHub at https://github.com/dbdimitrov/BingleSeq/.


2021 ◽  
Vol 2 (1) ◽  
pp. 43-61
Author(s):  
Aanchal Malhotra ◽  
Samarendra Das ◽  
Shesh N. Rai

Single-cell RNA-sequencing (scRNA-seq) technology provides an excellent platform for measuring the expression profiles of genes in heterogeneous cell populations. Multiple tools for the analysis of scRNA-seq data have been developed over the years. The tools require complicated commands and steps to analyze the underlying data, which are not easy to follow by genome researchers and experimental biologists. Therefore, we describe a step-by-step workflow for processing and analyzing the scRNA-seq unique molecular identifier (UMI) data from Human Lung Adenocarcinoma cell lines. We demonstrate the basic analyses including quality check, mapping and quantification of transcript abundance through suitable real data example to obtain UMI count data. Further, we performed basic statistical analyses, such as zero-inflation, differential expression and clustering analyses on the obtained count data. We studied the effects of excess zero-inflation present in scRNA-seq data on the downstream analyses. Our findings indicate that the zero-inflation associated with UMI data had no or minimal role in clustering, while it had significant effect on identifying differentially expressed genes. We also provide an insight into the comparative analysis for differential expression analysis tools based on zero-inflated negative binomial and negative binomial models on scRNA-seq data. The sensitivity analysis enhanced our findings in that the negative binomial model-based tool did not provide an accurate and efficient way to analyze the scRNA-seq data. This study provides a set of guidelines for the users to handle and analyze real scRNA-seq data more easily.


2017 ◽  
Vol 45 (19) ◽  
pp. 10978-10988 ◽  
Author(s):  
Cheng Jia ◽  
Yu Hu ◽  
Derek Kelly ◽  
Junhyong Kim ◽  
Mingyao Li ◽  
...  

Author(s):  
Donald Quicke ◽  
Buntika A. Butcher ◽  
Rachel Kruft Welton

Abstract This chapter employs generalized linear modelling using the function glm when we know that variances are not constant with one or more explanatory variables and/or we know that the errors cannot be normally distributed, for example, they may be binary data, or count data where negative values are impossible, or proportions which are constrained between 0 and 1. A glm seeks to determine how much of the variation in the response variable can be explained by each explanatory variable, and whether such relationships are statistically significant. The data for generalized linear models take the form of a continuous response variable and a combination of continuous and discrete explanatory variables.


PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1978 ◽  
Author(s):  
Gabriela Fontanarrosa ◽  
Virginia Abdala

Grasping is one of a few adaptive mechanisms that, in conjunction with clinging, hooking, arm swinging, adhering, and flying, allowed for incursion into the arboreal eco-space. Little research has been done that addresses grasping as an enhanced manual ability in non-mammalian tetrapods, with the exception of studies comparing the anatomy of muscle and tendon structure. Previous studies showed that grasping abilities allow exploitation for narrow branch habitats and that this adaptation has clear osteological consequences. The objective of this work is to ascertain the existence of morphometric descriptors in the hand skeleton of lizards related to grasping functionality. A morphological matrix was constructed using 51 morphometric variables in 278 specimens, from 24 genera and 13 families of Squamata. To reduce the dimensions of the dataset and to organize the original variables into a simpler system, three PCAs (Principal Component Analyses) were performed using the subsets of (1) carpal variables, (2) metacarpal variables, and (3) phalanges variables. The variables that demonstrated the most significant contributions to the construction of the PCA synthetic variables were then used in subsequent analyses. To explore which morphological variables better explain the variations in the functional setting, we ranGeneralized Linear Modelsfor the three different sets. This method allows us to model the morphology that enables a particular functional trait. Grasping was considered the only response variable, taking the value of 0 or 1, while the original variables retained by the PCAs were considered predictor variables. Our analyses yielded six variables associated with grasping abilities: two belong to the carpal bones, two belong to the metacarpals and two belong to the phalanges. Grasping in lizards can be performed with hands exhibiting at least two different independently originated combinations of bones. The first is a combination of a highly elongated centrale bone, reduced palmar sesamoid, divergence angles above 90°, and slender metacarpal V and phalanges, such as exhibited byAnolissp. andTropidurussp. The second includes an elongated centrale bone, lack of a palmar sesamoid, divergence angles above 90°, and narrow metacarpal V and phalanges, as exhibited by geckos. Our data suggest that the morphological distinction between graspers and non-graspers is demonstrating the existence of ranges along the morphological continuum within which a new ability is generated. Our results support the hypothesis of the nested origin of grasping abilities within arboreality. Thus, the manifestation of grasping abilities as a response to locomotive selective pressure in the context of narrow-branch eco-spaces could also enable other grasping-dependent biological roles, such as prey handling.


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