CellRank2 (Weiler et al, 2023) is a powerful framework for studying cellular fate using single-cell RNA sequencing data. It can handle millions of cells and different data types efficiently. This tool can identify cell fate and probabilities across various data sets. It also allows for analyzing transitions over time and uncovering key genes in developmental processes. Additionally, CellRank2 estimates cell-specific transcription and degradation rates, aiding in understanding differentiation trajectories and regulatory mechanisms.
In this notebook, we will use a primary tumor sample of patient T71 from the dataset GSE137804 (Dong R. et al, 2020) as an example. We have performed RNA-velocity analysis and pseudotime calculation on this dataset in scVelo (Bergen et al, 2020) notebook. The output will be then loaded into this CellRank2 notebook for further analysis.
This notebook is based on the tutorial provided on CellRank2 documentation. We have modified the notebook and changed the input data to show how the tool works on BioTuring's platform.
Single-cell RNA data allows cell-cell communications (***CCC***) methods to infer CCC at either the individual cell or cell cluster/cell type level, but physical distances between cells are not preserved Almet, Axel A., et al., (2021). On the other hand, spatial data provides spatial distances between cells, but single-cell or gene resolution is potentially lost. Therefore, integrating two types of data in a proper manner can complement their strengths and limitations, from that improve CCC analysis.
In this pipeline, we analyze CCC on Visium data with single-cell data as a reference. The pipeline includes 4 sub-notebooks as following
01-deconvolution: This step involves deconvolution and cell type annotation for Visium data, with cell type information obtained from a relevant single-cell dataset. The deconvolution method is SpatialDWLS which is integrated in Giotto package.
02-giotto: performs spatial based CCC and expression based CCC on Visium data using Giotto method.
03-nichenet: performs spatial based CCC and expression based CCC on Visium data using NicheNet method.
04-visualization: visualizes CCC results obtained from Giotto and NicheNet.
The development of large-scale single-cell atlases has allowed describing cell states in a more detailed manner. Meanwhile, current deep leanring methods enable rapid analysis of newly generated query datasets by mapping them into reference atlases.
expiMap (‘explainable programmable mapper’) Lotfollahi, Mohammad, et al. is one of the methods proposed for single-cell reference mapping. Furthermore, it incorporates prior knowledge from gene sets databases or users to analyze query data in the context of known gene programs (GPs).
scVI-tools (single-cell variational inference tools) is a package for end-to-end analysis of single-cell omics data primarily developed and maintained by the Yosef Lab at UC Berkeley. scvi-tools has two components
- Interface for easy use of a range of probabilistic models for single-cell omics (e.g., scVI, scANVI, totalVI).
- Tools to build new probabilistic models, which are powered by PyTorch, PyTorch Lightning, and Pyro.
Single-cell RNA-seq datasets in diverse biological and clinical conditions provide great opportunities for the full transcriptional characterization of cell types.
However, the integration of these datasets is challeging as they remain biological and techinical differences. **Harmony** is an algorithm allowing fast, sensitive and accurate single-cell data integration.