Spatial-HP-Skin

This repo will cover all of the code linked to Castillo et al.

(Manuscript Title: Spatial transcriptomics stratifies health and psoriatic disease severity by emergent cellular ecosystems)

WORKFLOW OVERVIEW

DEPENDENCIES

Please make sure to install all of the dependencies prior to running the code. The following dependencies are essential:-

CODE AND REPOSITORY NAVIGATION

The code is organised into several R notebooks with each notebook covering different parts of the analysis. The figure numbers and references are found within each notebook and a table content will also be added to link each figure to different parts of code for ease of navigation. Reproducibility is the key here, please contact XXXX for any issues with the code.

The notebooks will also require you to import the data from the linked Zenodo / Figshare repository (Will be added soon) in order to reproduce all of the respective figures.

Each notebook link will take you to the notebook with necessary code, please select the figure number in the table of contents found on top of the notebook to .

FIGURE 1

FIGURE NO NOTEBOOK LINK DESCRIPTION
1B link Spatial Plot - Healthy sample with cluster labels
1C link UMAP (Healthy samples only) - with spatial regions labels
1D link Percentage composition (Healthy samples only)
1E link Seurat Integeration (scRNA + ST data) showing cell type enrichment
1G link MIA - healthy skin with dataset 1 (Hughes et al.) - Structural cell types only
1H link MIA - healthy skin with dataset 2 (Reynolds et al.) - Structural cell types only
1I link MIA - healthy skin with dataset 3 (Sullivan et al.) - APCs and Preadiplocytes

FIGURE 2

FIGURE NO NOTEBOOK LINK DESCRIPTION
2A link MIA - healthy skin with dataset 1 (Hughes et al.) - Immune cell types only
2B link MIA - healthy skin with dataset 2 (Reynolds et al.) - Immune cell types only

FIGURE 3

FIGURE NO NOTEBOOK LINK DESCRIPTION
3A link Spatial Plots with cluster labels - Lesional and Non-Lesional skin
3B link UMAP - All samples combined (PS + Healthy skin) after Harmony batch correction
3C link Percentage composition plots for different clusters / spatial regions

FIGURE 4

FIGURE NO NOTEBOOK LINK DESCRIPTION
4E link Circos plot visualization - cluster 4 receptor with clusters 7, 3 and 10 as ligand.
4F link Circos plot visualization - cluster 7 receptor with clusters 10, 3 and 4 as ligand.
4G link Circos plot visualization - cluster 10 receptor with clusters 7, 3 and 4 as ligand.

FIGURE 5

FIGURE NO NOTEBOOK LINK DESCRIPTION
5A link MIA for (PS only) dataset 1 - Immune cell types
5B link MIA for (PS only) dataset 2 - Immune cell types
5C link MIA for TRM (Tisssue resident memory cells) dataset
5D link Space-fold projection - Healthy, non-lesional and lesional skin.
5E link B cell (From dataset 1) pathways

FIGURE 6

FIGURE NO NOTEBOOK LINK DESCRIPTION
6A link Hierarchical clustering based heatmap for all samples
6B link PCA plot for all samples - grouped by PS group
6C link PCA plot for all samples - grouped by Disease severity

FIGURE 7

FIGURE NO NOTEBOOK LINK DESCRIPTION
7A link Space-fold projection - Mild disease (split into non-lesional and lesional)
7B link Space-fold projection - Moderate-severe disease (split into non-lesional and lesional)

FIGURE 8

FIGURE NO NOTEBOOK LINK DESCRIPTION
8A link PCA plot - Pseudobulk for cluster 1 (Derm macrophages, fibroblasts)
8B link PCA plot - Pseudobulk for cluster 12 (Derm macrophages, fibroblasts)
8E link Spatial Feature plots #1

SUPPLEMENTARY FIGURE 2

FIGURE NO NOTEBOOK LINK DESCRIPTION
S2-B link Spatial Plots (Before and after filtering)
S2-C link Heat-map showing top 8 marker genes per cluster (Healthy sample data only)
S2-D link UMI counts

SUPPLEMENTARY FIGURE 3

FIGURE NO NOTEBOOK LINK DESCRIPTION
S3-A link Heatmap for Healthy samples - after Harmony batch correction
S3-B link UMAP - Healthy skin after Harmony batch correction
S3-C link Percentage composition plots for different clusters / spatial regions (Healthy skin)

SUPPLEMENTARY FIGURE 4

FIGURE NO NOTEBOOK LINK DESCRIPTION
S4-A link UMAP - healthy skin with scRNA dataset 1 (Hughes et al.)
S4-B link UMAP - healthy skin with scRNA dataset 2 (Reynolds et al.)

SUPPLEMENTARY FIGURE 5

FIGURE NO NOTEBOOK LINK DESCRIPTION
S5-A link (Anchor data UMAP) UMAP plots - Harmony batch correction vs Anchor batch correction
S5-A link (Harmony data UMAP) UMAP plots - Harmony batch correction vs Anchor batch correction
S5-C link (Anchor data) Spatial plots- Harmony batch correction vs Anchor batch correction
S5-C link (Harmony data) Spatial plots- Harmony batch correction vs Anchor batch correction

SUPPLEMENTARY FIGURE 6

FIGURE NO NOTEBOOK LINK DESCRIPTION
S6 link Heat-map showing top 10 marker genes per cluster (All samples) - Harmony batch corrected

SUPPLEMENTARY FIGURE 7

FIGURE NO NOTEBOOK LINK DESCRIPTION
S7 link Heat-map showing top 10 marker genes per cluster (All samples) - Anchor batch corrected

SUPPLEMENTARY FIGURE 8

FIGURE NO NOTEBOOK LINK DESCRIPTION
S8-B link UMIs per clusters - Harmony batch corrected data

SUPPLEMENTARY FIGURE 9

FIGURE NO NOTEBOOK LINK DESCRIPTION
S9-A link Seurat Integration (scRNA + ST data) showing cell type enrichment fpr psoriatic lesional skin ST
S9-B link Psoriatic skin scRNA-seq dataset 1
S9-C link Psoriatic skin scRNA-seq dataset 2
S9-D link MIA - PS skin with dataset 1 (Hughes et al.) - Structural cell types only
S9-E link MIA - PS skin with dataset 2 (Reynolds et al.) - Structural cell types only

SUPPLEMENTARY FIGURE 10

FIGURE NO NOTEBOOK LINK DESCRIPTION
S10A link Dendogram for all samples based on pseudo-bulk expression

SUPPLEMENTARY FIGURE 11

FIGURE NO NOTEBOOK LINK DESCRIPTION
S11 link PCA plots - per cluster basis (PS samples only)

SUPPLEMENTARY FIGURE 13

FIGURE NO NOTEBOOK LINK DESCRIPTION
S13 link Spatial Feature expression plots #2