E-spatial

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.

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.

Computational cell-type deconvolution is an important analytic technique for modeling the compositional heterogeneity of bulk gene expression data. A conceptually new Bayesian approach to this problem, BayesPrism, has recently been proposed and has subsequently been shown to be superior in accuracy and robustness against model misspecifications by independent studies. However, given that BayesPrism relies on Gibbs sampling, it is orders of magnitude more computationally expensive than standard approaches. InstaPrism is an R package for cell type composition and gene expression deconvolution in bulk RNA-Seq data. Based on the same conceptual Bayesian framework as BayesPrism, InstaPrism re-implements BayesPrism in a derandomized framework by replacing the time-consuming Gibbs sampling steps in BayesPrism with a fixed-point algorithm, which greatly accelerated the calculation speed while maintaining highly comparable performance. It works as an independent R package and does not require the users to have BayesPrism installed.

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.

Single-cell RNA sequencing (scRNA-seq) protocols often face challenges in measuring the expression of all genes within a cell due to various factors, such as technical noise, the sensitivity of scRNA-seq techniques, or sample quality. This limitation gives rise to a need for the prediction of unmeasured gene expression values (also known as dropout imputation) from scRNA-seq data. ADImpute (Leote A, 2023) is an R package combining several dropout imputation methods, including two existing methods (DrImpute, SAVER), two novel implementations: Network, a gene regulatory network-based approach using gene-gene relationships learned from external data, and Baseline, a method corresponding to a sample-wide average.. This notebook is to illustrate an example workflow of ADImpute on sample datasets loaded from the package. The notebook content is inspired from ADImpute's vignette and modified to demonstrate how the tool works on BioTuring's platform.

InferCNV is used to explore tumor single cell RNA-Seq data to identify evidence for somatic large-scale chromosomal copy number alterations, such as gains or deletions of entire chromosomes or large segments of chromosomes. This is done by exploring expression intensity of genes across positions of tumor genome in comparison to a set of reference 'normal' cells. A heatmap is generated illustrating the relative expression intensities across each chromosome, and it often becomes readily apparent as to which regions of the tumor genome are over-abundant or less-abundant as compared to that of normal cells. **Infercnvpy** is a scalable python library to infer copy number variation (CNV) events from single cell transcriptomics data. It is heavliy inspired by InferCNV, but plays nicely with scanpy and is much more scalable.

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.

Tumors are complex tissues of cancerous cells surrounded by a heterogeneous cellular microenvironment with which they interact. Single-cell sequencing enables molecular characterization of single cells within the tumor. However, cell annotation—the assignment of cell type or cell state to each sequenced cell—is a challenge, especially identifying tumor cells within single-cell or spatial sequencing experiments. Here, we propose ikarus, a machine learning pipeline aimed at distinguishing tumor cells from normal cells at the single-cell level. We test ikarus on multiple single-cell datasets, showing that it achieves high sensitivity and specificity in multiple experimental contexts. **InferCNV** is a Bayesian method, which agglomerates the expression signal of genomically adjointed genes to ascertain whether there is a gain or loss of a certain larger genomic segment. We have used **inferCNV** to call copy number variations in all samples used in the manuscript.

SCENIC Suite is a set of tools to study and decipher gene regulation. Its core is based on SCENIC (Single-Cell Regulatory Network Inference and Clustering) which enables you to infer transcription factors, gene regulatory networks and cell types from single-cell RNA-seq data. pySCENIC is a lightning-fast python implementation of the SCENIC pipeline (Single-Cell Regulatory Network Inference and Clustering) which enables biologists to infer transcription factors, gene regulatory networks and cell types from single-cell RNA-seq data.

This tool provides a user-friendly and automated way to analyze large-scale single-cell RNA-seq datasets stored in RDS (Seurat) format. It allows users to run various analysis tools on their data in one command, streamlining the analysis workflow and saving time. Note that this notebook is only for the demonstration of the tool. User can run the tool directly through the command line. Currently, we support: - InferCNV - Identifying tumor cells at the single-cell level using machine learning

In the realm of cancer research, grasping the intricacies of intratumor heterogeneity and its interplay with the immune system is paramount for deciphering treatment resistance and tumor progression. While single-cell RNA sequencing unveils diverse transcriptional programs, the challenge persists in automatically discerning malignant cells from non-malignant ones within complex datasets featuring varying coverage depths. Thus, there arises a compelling need for an automated solution to this classification conundrum. SCEVAN (De Falco et al., 2023), a variational algorithm, is designed to autonomously identify the clonal copy number substructure of tumors using single-cell data. It automatically separates malignant cells from non-malignant ones, and subsequently, groups of malignant cells are examined through an optimization-driven joint segmentation process.

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

The goal of this book is to teach newcomers and advanced professionals alike, the best practices of single-cell sequencing analysis. This book will teach you the most common analysis steps ranging from preprocessing to visualization to statistical evaluation and beyond. A read through the entire book will enable you to analyze unimodal and multimodal single-cell sequencing data on your own. The guidelines and recommendations in this book are not only tailored to teach you how to do single-cell analysis in general, but how to do them right. We base our suggestions on external benchmarks and reviews whenever possible. Finally, we consider this book to be a living resource for single-cell data analysts which can easily be updated when the recommendations change.

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.

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.