E-spatial

SCANPY integrates the analysis possibilities of established R-based frameworks and provides them in a scalable and modular form. Specifically, SCANPY provides preprocessing comparable to SEURAT and CELL RANGER, visualization through TSNE, graph-drawing and diffusion maps, clustering similar to PHENOGRAPH, identification of marker genes for clusters via differential expression tests and pseudotemporal ordering via diffusion pseudotime, which compares favorably with MONOCLE 2, and WISHBONE.

Geneformer is a foundation transformer model pretrained on a large-scale corpus of ~30 million single cell transcriptomes to enable context-aware predictions in settings with limited data in network biology. Here, we will demonstrate a basic workflow to work with ***Geneformer*** models. These notebooks include the instruction to: 1. Prepare input datasets 2. Finetune Geneformer model to perform specific task 3. Using finetuning models for cell classification and gene classification application

WGCNA: an R package for Weighted Gene Correlation Network Analysis Correlation networks are increasingly being used in bioinformatics applications. For example, weighted gene co-expression network analysis is a systems biology method for describing the correlation patterns among genes across microarray samples. Weighted correlation network analysis (WGCNA) can be used for: - Finding clusters (modules) of highly correlated genes - Summarizing such clusters using the module eigengene or an intramodular hub gene - Relating modules to one another and to external sample traits (using eigengene network methodology) - For calculating module membership measures All of these are important for identifying potential candidate genes associated with measured traits as well as identifying genes that are consistently co-expressed and could be contributing to similar molecular pathways. Using WGCNA is also extremely useful statistically as it accounts for inter-individual variation in gene expression and alleviates issues associated with multiple testing.

Understanding global communications among cells requires accurate representation of cell-cell signaling links and effective systems-level analyses of those links. We construct a database of interactions among ligands, receptors and their cofactors that accurately represent known heteromeric molecular complexes. We then develop **CellChat**, a tool that is able to quantitatively infer and analyze intercellular communication networks from single-cell RNA-sequencing (scRNA-seq) data. CellChat predicts major signaling inputs and outputs for cells and how those cells and signals coordinate for functions using network analysis and pattern recognition approaches. Through manifold learning and quantitative contrasts, CellChat classifies signaling pathways and delineates conserved and context-specific pathways across different datasets. Applying **CellChat** to mouse and human skin datasets shows its ability to extract complex signaling patterns.

This notebook illustrates how to convert data from a Seurat object into a Scanpy annotation data and a Scanpy annotation data into a Seurat object using the BioStudio data transformation library (currently under development). It facilitates continued research using libraries that interact with Scanpy in Python and Seurat in R. seurat.to.adata function can retain information about reductions (such as PCA, t-SNE, UMAP and Seurat Clusters) and spatial information.

Dynamic expression data, nowadays obtained using high-throughput RNA sequencing (RNA-seq), are essential to monitor transient gene expression changes and to study the dynamics of their transcriptional activity in the cell or response to stimuli. FunPat is an R package designed to provide: - a useful tool to analyze time series genomic data; - a computational pipeline which integrates gene selection, clustering and functional annotations into a single framework to identify the main temporal patterns associated to functional groups of differentially expressed (DE) genes; - an easy way to exploit different types of annotations from currently available databases (e.g. Gene Ontology) to extract the most meaningful information characterizing the main expression dynamics; - a user-friendly organization and visualization of the outcome, automatically linking the DE genes and their temporal patterns to the functional information for an easy biological interpretation of the results.

Build single-cell trajectories with the software that introduced **pseudotime**. Find out about cell fate decisions and the genes regulated as they're made. Group and classify your cells based on gene expression. Identify new cell types and states and the genes that distinguish them. Find genes that vary between cell types and states, over trajectories, or in response to perturbations using statistically robust, flexible differential analysis. In development, disease, and throughout life, cells transition from one state to another. Monocle introduced the concept of **pseudotime**, which is a measure of how far a cell has moved through biological progress. Many researchers are using single-cell RNA-Seq to discover new cell types. Monocle 3 can help you purify them or characterize them further by identifying key marker genes that you can use in follow-up experiments such as immunofluorescence or flow sorting. **Single-cell trajectory analysis** shows how cells choose between one of several possible end states. The new reconstruction algorithms introduced in Monocle 3 can robustly reveal branching trajectories, along with the genes that cells use to navigate these decisions.

In this notebook, we present COMMOT (COMMunication analysis by Optimal Transport) to infer cell-cell communication (CCC) in spatial transcriptomic, a package that infers CCC by simultaneously considering numerous ligand–receptor pairs for either spatial transcriptomic data or spatially annotated scRNA-seq data equipped with spatial distances between cells estimated from paired spatial imaging data. A collective optimal transport method is developed to handle complex molecular interactions and spatial constraints. Furthermore, we introduce downstream analysis tools to infer spatial signaling directionality and genes regulated by signaling using machine learning models.

Knowledge of cell type composition in disease relevant tissues is an important step towards the identification of cellular targets of disease. MuSiC is a method that utilizes cell-type specific gene expression from single-cell RNA sequencing (RNA-seq) data to characterize cell type compositions from bulk RNA-seq data in complex tissues. By appropriate weighting of genes showing cross-subject and cross-cell consistency, MuSiC enables the transfer of cell type-specific gene expression information from one dataset to another. MuSiC enables the characterization of cellular heterogeneity of complex tissues for understanding of disease mechanisms. As bulk tissue data are more easily accessible than single-cell RNA-seq, MuSiC allows the utilization of the vast amounts of disease relevant bulk tissue RNA-seq data for elucidating cell type contributions in disease. This notebook provides a walk through tutorial on how to use MuSiC to estimate cell type proportions from bulk sequencing data based on multi-subject single cell data by reproducing the analysis in MuSiC paper, now is published on Nature Communications.

Monorail can be used to process local and/or private data, allowing results to be directly compared to any study in recount3. Taken together, Monorail-pipeline tools help biologists maximize the utility of publicly available RNA-seq data, especially to improve their understanding of newly collected data. This is for helping potential users of the Monorail RNA-seq processing pipeline (alignment/quantification) get started running their own data through it.

Dampened weighted least squares (DWLS) is an estimation method for gene expression deconvolution, in which the cell-type composition of a bulk RNA-seq data set is computationally inferred. This method corrects common biases towards cell types that are characterized by highly expressed genes and/or are highly prevalent, to provide accurate detection across diverse cell types. To begin, the user must input a bulk RNA-seq data set, along with a labeled representative single-cell RNA-seq data set that will serve to generate cell-type-specific gene expression profiles. Ideally, the single-cell data set will contain cells from all cell types that may be found in the bulk data. DWLS will return the cell-type composition of the bulk data.

BPCells is a package for high performance single cell analysis on RNA-seq and ATAC-seq datasets. It can analyze a 1.3M cell dataset with 2GB of RAM in under 10 minutes. This makes analysis of million-cell datasets practical on a laptop. BPCells provides: * Efficient storage of single cell datasets via bitpacking compression * Fast, disk-backed RNA-seq and ATAC-seq data processing powered by C++ * Downstream analysis such as marker genes, and clustering * Interoperability with AnnData, 10x datasets, R sparse matrices, and GRanges

The simultaneous measurement of multiple modalities represents an exciting frontier for single-cell genomics and necessitates computational methods that can define cellular states based on multimodal data. Here, we introduce "weighted-nearest neighbor" analysis, an unsupervised framework to learn the relative utility of each data type in each cell, enabling an integrative analysis of multiple modalities. We apply our procedure to a CITE-seq dataset of 211,000 human peripheral blood mononuclear cells (PBMCs) with panels extending to 228 antibodies to construct a multimodal reference atlas of the circulating immune system. Multimodal analysis substantially improves our ability to resolve cell states, allowing us to identify and validate previously unreported lymphoid subpopulations. Moreover, we demonstrate how to leverage this reference to rapidly map new datasets and to interpret immune responses to vaccination and coronavirus disease 2019 (COVID-19). Our approach represents a broadly applicable strategy to analyze single-cell multimodal datasets and to look beyond the transcriptome toward a unified and multimodal definition of cellular identity.

Single-cell RNA sequencing (scRNA-seq) has revolutionized the study of gene expression at the individual cell level, enabling researchers to uncover heterogeneity and dynamics within complex cellular populations. To analyze and interpret scRNA-seq data effectively, bioinformaticians often rely on specialized tools. In this benchmarking study, we aim to compare the performance of Bioturing Alpha, Scanpy and Seurat in terms of their execution time in various tools of scRNA-seq analysis pipeline. Tools used in scRNA-seq analysis pipeline: - Preprocessing: This step involves tasks such as quality control, filtering out low-quality cells, normalizing gene expression, identifying highly variable genes and regress out unwanted variance. - Linear Dimensionality Reduction: Perform linear dimensionality reduction using techniques such as Principal Component Analysis (PCA) before applying batch effect removal. PCA captures the most significant sources of variation in the data by projecting it onto orthogonal axes. - Batch Effect Removal: Apply batch effect removal methods using Harmony integrate to mitigate the influence of batch effects on downstream analyses. - Clustering: Apply clustering algorithms to partition cells into distinct clusters based on their gene expression profiles. Evaluate the accuracy of cluster assignments by comparing them to known cell types, if available. - Non-Linear Dimensionality Reduction: Utilize non-linear dimensionality reduction techniques, such as t-Distributed Stochastic Neighbor Embedding (t-SNE) and Uniform Manifold Approximation and Projection (UMAP). Apply t-SNE and UMAP to visualize the data in lower-dimensional spaces and assess their ability to reveal intricate cluster patterns.

Generative pre-trained models have demonstrated exceptional success in various fields, including natural language processing and computer vision. In line with this progress, scGPT has been developed as a foundational model tailored specifically for the field of single-cell biology. It employs the generative pre-training transformer framework on an extensive dataset comprising more than 33 million cells. scGPT effectively extracts valuable biological insights related to genes and cells and can be fine-tuned to excel in numerous downstream applications.