--- title: "How to interpret the HTML report generated by `cellCellReport` function" author: - name: Koki Tsuyuzaki affiliation: Laboratory for Bioinformatics Research, RIKEN Center for Biosystems Dynamics Research - name: Manabu Ishii affiliation: Laboratory for Bioinformatics Research, RIKEN Center for Biosystems Dynamics Research - name: Itoshi Nikaido affiliation: Laboratory for Bioinformatics Research, RIKEN Center for Biosystems Dynamics Research email: k.t.the-answer@hotmail.co.jp package: scTensor output: BiocStyle::html_document vignette: | %\VignetteIndexEntry{scTensor: 2. Interpretation of HTML report} %\VignetteEngine{knitr::rmarkdown} %\VignetteEncoding{UTF-8} --- # Introduction Here, we explain the way to interpret of HTML report generated by `cellCellReport`. If `cellCellDecomp` is properly finished, we can perform `cellCellReport` function to output the HTML report. The results can be confirmed by typing `example(cellCellReport)`. The report will be generated in the temporary directory (it costs 5 to 10 minutes). The output directory contains some files and directories as follows. - **index.{Rmd,html}** : The main HTML report - **reanalysis.RData** : The R binary file for using in the HTML report and reanalysis of scTensor - **Workflow_2.png** : The figure for the section 1. About scTensor Algorithm in the HTML report - **figures** : The directory containing some figures for the HTML report - **ligand.{Rmd,html}** : The HTML report for the section 6. Gene-wise Hypergraph in the HTML report - **ligand_all.{Rmd,html}** : The HTML report for the section 6. Gene-wise Hypergraph in the HTML report - **receptor.{Rmd,html}** : The HTML report for the section 6. Gene-wise Hypergraph in the HTML report - **receptor_all.{Rmd,html}** : The HTML report for the section 6. Gene-wise Hypergraph in the HTML report - **pattern_X_Y.{Rmd,html}** : The HTML report for For the section 7. (Ligand-Cell, Receptor-Cell, ) -related L-R Pairs in the HTML report Here, look at the index.html. ![Figure1 : HTML report of `cellCellReport`](Report_HEADER.png) # Interpretation of "1. About scTensor Algorithm" In the HTML report, the 1st item describes the overview of `r Biocpkg("scTensor")` and other CCI-related packages. ![Figure2: 1. About scTensor Algorithm](Report_1.png) # Interpretation of "2. Global statistics and plots" The 2nd item describes all the R objects saved in **reanalysis.RData**, which contains the result of `r Biocpkg("scTensor")`. This file is saved in the output directory (**out.dir**) specified in `cellCellReport`, and the user also can re-analyze the result of `r Biocpkg("scTensor")`. ![Figure3: 2. Global statistics and plots](Report_2.jpg) Using `r CRANpkg("plotly")` package, `cellCellReport` generates some interactive plots. For example, in item 2.1, the number of cells in each cell type can be confirmed when the cursor moved on the box. ![Figure4: 2.1 Number of cells in each celltype](Report_2_1.jpg) In item 2.2, the number of expressed genes in each cell type (Non-zero genes) can be confirmed when the cursor moved on the box. ![Figure5: 2.2 Number of expressed genes in each cell type (Non-zero genes)](Report_2_2.jpg) In item 2.3, the two-dimensional plot user specified can be confirmed. ![Figure6: 2.3 Two dimensional plot of all cells](Report_2_3.jpg) In item 2.4, the distribution of core tensor values and the value of each (Ligand-Cell-type, Receptor-Cell-type, LR-pair) pattern can be confirmed. The red bars mean that these values are selected by the threshold (**thr** parameters) in `cellCellReport`. Note that the thr can be specified from 0 to 100, the large thr value will generate too many HTML files (cf. 8. (Ligand-Cell, Receptor-Cell, LR-pair) Patterns) and takes a long time. ![Figure7 : 2.4 Distribution of core tensor values](Report_2_4.png) The 3-order CCI-tensor consisting of Cell_L $\times$ Cell_R $\times$ LR-pair (LR) are decomposed by `r CRANpkg("nnTensor")`, in which the tensor is iteratively matricised to mode-1 (Ligand-Cell direction) and mode-2 (Receptor-Cell direction). In each direction, NMF is performed and the strength of each directional pattern is summarized in the bar plots. For example, in item 2.5, the distribution of mode-1 matricised tensor can be confirmed. ![Figure8: 2.5 Distribution of mode-1 matricised tensor (Ligand-Cell Direction) (1/2)](Mode1.jpg) ![Figure9: 2.5 Distribution of mode-1 matricised tensor (Ligand-Cell Direction) (2/2)](Report_2_5.jpg) Likewise, in item 2.6, the distribution of mode-2 matricised tensor can be confirmed, ![Figure10 : 2.6 Distribution of mode-2 matricised tensor (Receptor-Cell Direction) (1/2)](Mode2.jpg) ![Figure11 : 2.6 Distribution of mode-2 matricised tensor (Receptor-Cell Direction) (2/2)](Report_2_6.jpg) # Interpretation of "3. Ligand-Cell Patterns" In the 3rd item, using the heatmap of `r CRANpkg("plotly")`, the user can interactively confirm the detail of Ligand-Cell Patterns extracted by `r CRANpkg("nnTensor")`. ![Figure14 : 3. Ligand-Cell Patterns](Report_3.jpg) # Interpretation of "4. Receptor-Cell Patterns" Likewise, in the 4th item, the user can interactively confirm the detail of Receptor-Cell Patterns. ![Figure15 : 4. Receptor-Cell Patterns](Report_4.jpg) # Interpretation of "5. CCI-wise Hypergraph" In the 6th item describes, the strength between Ligand-Cell Patterns and Receptor-Cell Patterns (CCI-strength), by the summation of the core tensor with the mode-3 direction, a matrix consisting of the number of Ligand-Cell Patterns $\times$ the number of Receptor-Cell Patterns. ![Figure18 : 6. CCI-wise Hypergraph (1/2)](Mode3Sum.jpg) ![Figure19 : 6. CCI-wise Hypergraph (2/2)](Report_5.png) # Interpretation of "6. Gene-wise Hypergraph" In the 7th item, the relationship between LR-pairs, which coexpressed in any LR-pair pattern at least one time. Ligand genes are described as red nodes, receptor genes are described as blue nodes, and corresponding LR-pair patterns are described as the color of edges. Using `r CRANpkg("visNetwork")` package, these interactions can be interactively visualized. ![Figure20 : 7. Gene-wise Hypergraph](Report_6.png) Under the gene-wise hypergraph, four hyperlinks are embedded. In the 1st link, the details of the gene-wise hypergraph can be confirmed as a corresponding table in a ligand gene-centric manner. This page can work as a reverse lookup search by "Ctrl + F"; by typing the gene name of ligand that the user is interested in, the partner receptors, which are coexpressed in some LR-pair patterns, also can be found. ![Figure21: Details of Ligand Gene-centric Overview (selected)](Ligand_selected.png) In the 2nd link, the user can find all the partner receptors, even if the partner receptors are not coexpressed in any LR-pair pattern, and if they are not included in the data matrix. ![Figure22: Details of Ligand Gene-centric Overview (all)](Ligand_all.png) Likewise, the receptor gene-centric reverse search page is embedded in the 3rd link, ![Figure23: Details of Receptor Gene-centric Overview (selected)](Receptor_selected.png) and, in the 4th link, all the partner ligand genes are included. ![Figure24: Details of Receptor Gene-centric Overview (all) (1/2)](Receptor_all_HEADER.png) ![Figure25 : Details of Receptor Gene-centric Overview (all) (2/2)](Receptor_all.png) # Interpretation of "7. (Ligand-Cell, Receptor-Cell, LR-pair) Patterns" In the 8th item, the details of (Ligand-Cell, Receptor-Cell, LR-pair) Patterns are ordered by the size of the core tensor, and the link of each pattern is embedded. (Note that the number of links is dependent on the **thr** parameter of `cellCellReport`.) ![Figure26: 8. (Ligand-Cell, Receptor-Cell, LR-pair) Patterns](Report_7.png) For example, the 1st link describes the details of (3,2,) Pattern, which means the relationship of *1*st pattern of Ligand-Cell patterns, *1*st pattern of Receptor-Cell patterns, and *5*th pattern of LR-pair patterns. ![Figure27 : Details of (3,2,) Pattern (1/3)](Details_32_HEADER.png) In this pattern, only one LR-pair is coexpressed (INSL3 and GNG11). The hyperlinks to many databases and PubMed are also embedded. The degree of the size of the LR-pair in the LR-pair pattern is quantified as P-value and Q-value. ![Figure28 : Details of (3,2,) Pattern (2/3)](Details_32_Pair.png) Under the LR-pair list, the results of many enrichment analysis are also embedded such as Gene Ontology (BP/MF/CC), Reactome, MeSH...etc. ![Figure29 : Details of (3,2,) Pattern (3/3)](Details_32_EA_HEADER.png) User can confirm the detail of the result of `r Biocpkg("scTensor")`, and perform the biological interpretation. # Session information {.unnumbered} ```{r sessionInfo, echo=FALSE} sessionInfo() ```