An integrated cell atlas of the lung in health and disease

Single-cell technologies have transformed our understanding of human tissues. Yet, studies typically capture only a limited number of donors and disagree on cell type definitions. Integrating many single-cell datasets can address these limitations of individual studies and capture the variability present in the population. Here we present the integrated Human Lung Cell Atlas (HLCA), combining 49 datasets of the human respiratory system into a single atlas spanning over 2.4 million cells from 486 individuals. The HLCA presents a consensus cell type re-annotation with matching marker genes, including annotations of rare and previously undescribed cell types. Leveraging the number and diversity of individuals in the HLCA, we identify gene modules that are associated with demographic covariates such as age, sex and body mass index, as well as gene modules changing expression along the proximal-to-distal axis of the bronchial tree. Mapping new data to the HLCA enables rapid data annotation and interpretation. Using the HLCA as a reference for the study of disease, we identify shared cell states across multiple lung diseases, including SPP1 + profibrotic monocyte-derived macrophages in COVID-19, pulmonary fibrosis and lung carcinoma. Overall, the HLCA serves as an example for the development and use of large-scale, cross-dataset organ atlases within the Human Cell Atlas. © 2023, The Author(s)

An integrated cell atlas of the lung in health and disease

Single-cell technologies have transformed our understanding of human tissues. Yet, studies typically capture only a limited number of donors and disagree on cell type definitions. Integrating many single-cell datasets can address these limitations of individual studies and capture the variability present in the population. Here we present the integrated Human Lung Cell Atlas (HLCA), combining 49 datasets of the human respiratory system into a single atlas spanning over 2.4 million cells from 486 individuals. The HLCA presents a consensus cell type re-annotation with matching marker genes, including annotations of rare and previously undescribed cell types. Leveraging the number and diversity of individuals in the HLCA, we identify gene modules that are associated with demographic covariates such as age, sex and body mass index, as well as gene modules changing expression along the proximal-to-distal axis of the bronchial tree. Mapping new data to the HLCA enables rapid data annotation and interpretation. Using the HLCA as a reference for the study of disease, we identify shared cell states across multiple lung diseases, including SPP1 + profibrotic monocyte-derived macrophages in COVID-19, pulmonary fibrosis and lung carcinoma. Overall, the HLCA serves as an example for the development and use of large-scale, cross-dataset organ atlases within the Human Cell Atlas. </p

An integrated cell atlas of the lung in health and disease

Single-cell technologies have transformed our understanding of human tissues. Yet, studies typically capture only a limited number of donors and disagree on cell type definitions. Integrating many single-cell datasets can address these limitations of individual studies and capture the variability present in the population. Here we present the integrated Human Lung Cell Atlas (HLCA), combining 49 datasets of the human respiratory system into a single atlas spanning over 2.4 million cells from 486 individuals. The HLCA presents a consensus cell type re-annotation with matching marker genes, including annotations of rare and previously undescribed cell types. Leveraging the number and diversity of individuals in the HLCA, we identify gene modules that are associated with demographic covariates such as age, sex and body mass index, as well as gene modules changing expression along the proximal-to-distal axis of the bronchial tree. Mapping new data to the HLCA enables rapid data annotation and interpretation. Using the HLCA as a reference for the study of disease, we identify shared cell states across multiple lung diseases, including SPP1+ profibrotic monocyte-derived macrophages in COVID-19, pulmonary fibrosis and lung carcinoma. Overall, the HLCA serves as an example for the development and use of large-scale, cross-dataset organ atlases within the Human Cell Atlas

Tools and techniques for single-cell RNA sequencing data

© 2019 Dr. Luke ZappiaRNA sequencing of individual cells allows us to take a snapshot of the dynamic processes within a cell and explore differences between cell types. As this technology has developed over the last few years it has been rapidly adopted by researchers in areas such as developmental biology, and many single-cell RNA sequencing datasets are now available. Coinciding with the development of protocols for producing single-cell RNA sequencing data there has been a simultaneous burst in the development of computational analysis methods. My thesis explores the computational tools and techniques for analysing single-cell RNA sequencing data. I present a database that charts the release of analysis software, where it has been published and what it can be used for, as well as a website that makes this information publicly available. I also present two of my own tools and techniques including Splatter, a software package for easily simulating single-cell datasets from multiple models, and clustering trees, a visualisation approach for inspecting clustering at multiple resolutions. In the final part of my thesis I perform analysis of a dataset from kidney organoids to demonstrate and compare some current analysis methods. Taken together, my thesis covers many aspects of the tools and techniques for single-cell RNA sequencing by describing the approaches that are available, presenting software that can help in developing and evaluating methods, introducing an approach for aiding one of the most common analysis tasks, and showing how tools can be used to extract meaning from a real dataset

Oshlack/splatter: Version 1.1.3 (BASiCS)

Version 1.1.3 of Splatter with the BASiCS simulation. This is the version of Splatter used in the response to reviewers for the Splatter paper https://doi.org/10.5281/zenodo.833572

Oshlack/splatter: Version 1.2.0

Bioconductor 3.6 release versio

Exploring the single-cell RNA-seq analysis landscape with the scRNA-tools database.

As single-cell RNA-sequencing (scRNA-seq) datasets have become more widespread the number of tools designed to analyse these data has dramatically increased. Navigating the vast sea of tools now available is becoming increasingly challenging for researchers. In order to better facilitate selection of appropriate analysis tools we have created the scRNA-tools database (www.scRNA-tools.org) to catalogue and curate analysis tools as they become available. Our database collects a range of information on each scRNA-seq analysis tool and categorises them according to the analysis tasks they perform. Exploration of this database gives insights into the areas of rapid development of analysis methods for scRNA-seq data. We see that many tools perform tasks specific to scRNA-seq analysis, particularly clustering and ordering of cells. We also find that the scRNA-seq community embraces an open-source and open-science approach, with most tools available under open-source licenses and preprints being extensively used as a means to describe methods. The scRNA-tools database provides a valuable resource for researchers embarking on scRNA-seq analysis and records the growth of the field over time

Splatter: simulation of single-cell RNA sequencing data

Abstract As single-cell RNA sequencing (scRNA-seq) technologies have rapidly developed, so have analysis methods. Many methods have been tested, developed, and validated using simulated datasets. Unfortunately, current simulations are often poorly documented, their similarity to real data is not demonstrated, or reproducible code is not available. Here, we present the Splatter Bioconductor package for simple, reproducible, and well-documented simulation of scRNA-seq data. Splatter provides an interface to multiple simulation methods including Splat, our own simulation, based on a gamma-Poisson distribution. Splat can simulate single populations of cells, populations with multiple cell types, or differentiation paths

Additional file 1: Figures S1–S17 of Splatter: simulation of single-cell RNA sequencing data

Diagrams of other simulation models, Splatter comparison output for all datasets, example non-linear gene, dispersion estimate correction, mean-zeros fit, benchmarking, and processing times (PDF 17991 kb

Exploring the single-cell RNA-seq analysis landscape with the scRNA-tools database - Fig 3

<p>(A) Categories of tools in the scRNA-tools database. Each tool can be assigned to multiple categories based on the tasks it can complete. Categories associated with multiple analysis phases (visualisation, dimensionality reduction) are among the most common, as are categories associated with the cell assignment phase (ordering, clustering). (B) Changes in analysis categories over time, comparing tools added before and after October 2016. There have been significant increases in the percentage of tools associated with visualisation, dimensionality reduction, gene networks and simulation. Categories including expression patterns, ordering and interactivity have seen relative decreases. (C) Changes in the percentage of tools associated with analysis phases over time. The percentage of tools involved in the data acquisition and data cleaning phases have increased, as have tools designed for alternative analysis tasks. The gene identification phase has seen a relative decrease in the number of tools. (D) The number of categories associated with each tools in the scRNA-tools database. The majority of tools perform few tasks. (E) Most tools that complete many tasks are relatively recent.</p