StereoSiTE is a package for the analysis and visualization of spatial transcriptome data. It builds on top of anndata, scanpy and squidpy, from which it inherits modularity and scalability. It provides analysis tools to dissect cellular neighborhood based on cell composition and quantitatively define cell-cell communication in spatial.
- Cellular Neighborhood (CN) clustering based on cellular composition of each bin
- Spatial Cell Interaction Intensity (SCII) analysis
- Tissue domain clustering based on intercellular interactions within each bin (SCIITensor)
demo (Fetch code: nFks)
If you use stereosite in your work, please cite the publication as follows:
StereoSiTE: A framework to spatially and quantitatively profile the cellular neighborhood organized iTME
Xing Liu, Chi Qu, Chuandong Liu, Na Zhu, Huaqiang Huang, Fei Teng, Caili Huang, Bingying Luo, Xuanzhu Liu, Yisong Xu, Min Xie, Feng Xi, Mei Li, Liang Wu, Yuxiang Li, Ao Chen, Xun Xu, Sha Liao, Jiajun Zhang bioRxiv 2022.12.31.522366; doi: https://doi.org/10.1101/2022.12.31.522366
SCIITensor: A tensor decomposition based algorithm to construct actionable TME modules with spatially resolved intercellular communications
Huaqiang Huang, Chuandong Liu, Xin Liu, Jingyi Tian, Feng Xi, Mei Li, Guibo Li, Ao Chen, Xun Xu, Sha Liao, Jiajun Zhang, Xing Liu bioRxiv 2024.05.21.595103; doi: https://doi.org/10.1101/2024.05.21.595103
Install StereoSiTE via PyPi by running:
>>> conda create -y -n stereosite python=3.9.12 >>> conda activate stereosite >>> pip install stereosite
or via raw code by running:
>>> git clone https://github.com/STOmics/StereoSiTE.git >>> cd StereoSiTE >>> python setup.py install
Transfer spatial gene expression matrix (gem) to anndata
>>> from stereosite.read.gem import Gem_Reader >>> gem_file = "to/path/sn.gem.gz" >>> gem_reader = Gem_Reader(gem_file) # without cell mask, bin_size must be specified. Here bin_size=200 >>> adata_bin200 = gem_reader.gem2anndata(200) >>> adata_bin200_file = "to/path/sn_bin200.h5ad" >>> adata_bin200.write(adata_bin200_file) # or with cell mask, gem will be transfered to anndata in single-cell resolution. >>> mask_file = "to/path/sn_mask.tif" >>> adata = gem_reader.gem_with_cellmask_2anndata(mask_file)
Cellular Neighborhood (CN)
>>> from stereosite.cn.deconvolution import Cell2location # Get the cellular composition of each square bin by deconvolution >>> ref_file = "to/path/scCell_reference.csv" >>> adata_file = "to/path/sn_bin200.h5ad" >>> out_dir = "to/out/deconvolution/bin200" >>> cell2loc = Cell2location(ref_file, adata_file, out_dir = out_dir, bin_size = 200, gpu = 0) >>> cell2loc.run_deconvolution() # Analyze the Cellular Neighborhood based on deconvolution result >>> from stereosite.cn.cellneighbor import cn_deconvolve >>> import anndata >>> adata_anno_file = "to/out/deconvolution/bin200/cell2location_map/sp.h5ad" >>> adata = anndata.read(adata_anno_file) # use_rep specify matrix used to calculate cell composition of every bin >>> cn_deconvolve(adata, use_rep='q05_cell_abundance_w_sf') # Or use the annotated cell bin data to dissect the CNs >>> from stereosite.cn.cellneighbor import cn_cellbin >>> import anndata >>> adata = anndata.read(adata_anno_file) >>> cn_cellbin(adata, 400, n_neighbors = 20, resolution = 0.4, min_dist = 0.1) # CN result visualization >>> from stereosite.plot.cellneighbor import umap, heatmap, spatial >>> spatial(adata, spot_size=20) >>> umap(adata) >>> heatmap(adata)
Spatial Cell Interaction Intensity (SCII)
>>> from stereosite.scii import intensities_count # The annotated cellbin or square bin at single-cell resolution data is required. # Choose LR database based on the sample type. CellChatDB provide mouse and human database separatly. # CellphoneDB provide only human database, but also can be used to analyse mouse data by transfer mouse gene into homologous # human gene, which will be automaticaly done by the software. >>> interactiondb_file = "./datasets/LR_database/CellChatDB.mouse.csv" >>> scii_dict = intensities_count(adata, interactiondb_file, >>> distance_threshold = 50, >>> anno = 'cell_type') # Or we can specify different distance_threshold for individual LR types >>> scii_dict = intensities_count(adata, interactiondb_file, >>> distance_threshold = {'Secreted Signaling': 200, 'ECM-Receptor': 200, 'Cell-Cell Contact': 30}, >>> anno = 'cell_type') # Or specify the distance_coefficient parameter to consider distance when caculating interaction intensity. # distance_coefficient=0 means distance would not influence the interaction intensity. >>> scii_dict = intensities_count(adata, interactiondb_file, >>> distance_threshold = {'Secreted Signaling': 200, 'ECM-Receptor': 200, 'Cell-Cell Contact': 30}, >>> distance_coefficient = {'Secreted Signaling': 1, 'ECM-Receptor': 0.1, 'Cell-Cell Contact': 0}, >>> anno = 'cell_type') # The interaction result can be writen into a pickle file, and can be re-loaded when you want to re-analyze it. >>> import pickle >>> os.makedirs("./out/scii", exist_ok=True) >>> interaction_file = "./out/scii/interactions.pkl" >>> with open(interaction_file, 'wb') as writer: >>> pickle.dump(scii_dict, writer) >>> with open(interaction_file, 'rb') as reader: >>> scii_dict = pickle.load(reader) # SCII result visualization >>> from stereosite.plot.scii import ligrec >>> import numpy as np >>> ligrec(scii_dict, >>> intensities_range=(50, np.inf), >>> pvalue_threshold=0.05, >>> anno="cell_type", >>> alpha=1e-4, >>> swap_axes=False, >>> source_groups=["Non-immune cells", "M2-like", 'DC', 'Teff'], >>> target_groups = ["M1-like", "M2-like", "Monocytes", "Teff", "CD8+ Tcells"], >>> title=" ", >>> ) # Show spatial distribution of interaction intensity between specific cell pair meidated by specific LR pair. >>> from stereosite.plot.intensity import intensity_insitu >>> cells = ['Non-immune cells', 'M2-like'] >>> genes = ['Ptprc', 'Mrc1'] >>> intensity_insitu(adata, cells, genes, radius = 50, distance_coefficient=0.01, spot_size=5)
SCIITensor -- single sample analysis
>>> from stereosite import scii_tensor >>> import anndata >>> import pandas as pd >>> import seaborn as sns >>> import matplotlib as mpl >>> import matplotlib.pyplot as plt >>> import pickle >>> import numpy as np >>> import scanpy as sc # Generate interactiontensor object and evaluate the optimal combination of ranks >>> adata = anndata.read(adata_anno_file) >>> interactionDB = "./datasets/LR_database/CellChatDB.mouse.csv" >>> sct = scii_tensor.InteractionTensor(adata, interactionDB=interactionDB) >>> radius = {'Secreted Signaling': 100, 'ECM-Receptor': 100, 'Cell-Cell Contact': 30} >>> scii_tensor.build_SCII(sct, radius=radius, window_size=200, anno_col='cell2loc_anno') >>> scii_tensor.process_SCII(sct, zero_remove=True, log_data=True) >>> reconstruction_errors = scii_tensor.evaluate_ranks(sct, use_gpu=True, device='cuda:1') # Visualize the reconstruction errors using line plot >>> from stereosite.plot.scii_tensor import reconstruction_error_line >>> reconstruction_error_line(reconstruction_errors, figsize=(4, 4)) # Decompose the interaction tensor with optimal combination of ranks >>> scii_tensor.SCII_Tensor(sct, rank=[15, 15, 15], device='cuda:0') >>> with open("out/scii_tensor_res.pkl", "wb") as f: >>> pickle.dump(sct, f) # spatial distribution of each TME module >>> import scanpy as sc >>> sc.pl.spatial(sct.adata, color='TME_module', img_key=None, spot_size=20) ## core heatmap >>> from stereosite.plot.scii_tensor import tme_core_heatmap >>> tme_core_heatmap(sct.core, tme_number=1, figsize=(4, 4)) ## cell-cell factor heatmap >>> from stereosite.scii_tensor import top_pair >>> import seaborn as sns >>> import matplotlib.pyplot as plt >>> top_cc_pair = top_pair(sct, pair='cc', top_n=20) >>> fig = sns.clustermap(top_cc_pair.T, cmap="Purples", standard_scale=0, metric='euclidean', method='ward', >>> row_cluster=False, dendrogram_ratio=0.05, cbar_pos=(1.02, 0.6, 0.01, 0.3), >>> figsize=(4, 6), >>> ) ## ligand-receptor factor heatmap >>> top_lr_pair = top_pair(sct, pair='lr', top_n=20) >>> fig = sns.clustermap(top_lr_pair.T, cmap="Purples", standard_scale=0, metric='euclidean', method='ward', >>> row_cluster=False, dendrogram_ratio=0.05, cbar_pos=(1.02, 0.6, 0.01, 0.3), >>> figsize=(4, 6), >>> ) ## visualize selected interactions using heatmap >>> from stereosite.plot.scii_tensor import interaction_heatmap >>> interactions = scii_tensor.interaction_select(sct, >>> tme_module=1, >>> cellpair_module=1, >>> lrpair_module=11, n_lr=15, n_cc=15) >>> interaction_heatmap(interactions, figsize=(5, 3), vmax=50) ## visualize selected interactions using circle plot >>> from stereosite.plot.scii_circos import cells_lr_circos, cells_circos, lr_circos >>> cells = adata.obs['cell2loc_anno'].unique() >>> cells_lr_circos(interactions, cells=cells) ## sankey >>> from stereosite.plot import sankey >>> core_df = sankey.core_process(sct.core) >>> sankey.sankey_3d(core_df, link_alpha=0.5, interval=0.001)
SCIITensor -- multiple sample analysis
>>> adata_1 = anndata.read(adata_anno_file_1) ## decompose another sample data ## evaluate the optimal combination of ranks >>> interactionDB = "./datasets/LR_database/CellChatDB.mouse.csv" >>> sct_1 = scii_tensor.InteractionTensor(adata_1, interactionDB=interactionDB) >>> radius = {'Secreted Signaling': 100, 'ECM-Receptor': 100, 'Cell-Cell Contact': 30} >>> scii_tensor.build_SCII(sct_1, radius=radius, window_size=200, anno_col='cell2loc_anno') >>> scii_tensor.process_SCII(sct_1, zero_remove=True, log_data=True) >>> reconstruction_errors = scii_tensor.evaluate_ranks(sct_1, use_gpu=True, device='cuda:1') ## visualize the reconstruction errors using line plot >>> from stereosite.plot.scii_tensor import reconstruction_error_line >>> reconstruction_error_line(reconstruction_errors, figsize=(4, 4)) >>> scii_tensor.SCII_Tensor(sct_1, rank=(20, 20, 13), device='cuda:0') >>> with open("out/scii_tensor_res_1.pkl", "wb") as f: >>> pickle.dump(sct_1, f) ## merge decomposed matrices >>> sct_merge = scii_tensor.merge_data([sct, sct_1], patient_id=['p1' ,'p2']) ## visualize the reconstruction errors >>> from stereosite.plot.scii_tensor import reconstruction_error_line >>> reconstruction_error_line(reconstruction_errors, figsize=(4, 4)) >>> scii_tensor.SCII_Tensor_multiple(sct_merge, rank=[15,15,10], device='cuda:1') ## spatial distribution of meta-module >>> sc.pl.spatial(sct_merge.adata[0], color=['TME_module', 'TME_meta_module'], img_key=None, spot_size=20) >>> sc.pl.spatial(sct_merge.adata[1], color=['TME_module', 'TME_meta_module'], img_key=None, spot_size=20) ## core heatmap >>> from stereosite.plot.scii_tensor import tme_core_heatmap >>> tme_core_heatmap(sct_merge.core, tme_number=0, figsize=(4, 3)) ## visualize selected interactions using heatmap >>> from stereosite.plot.scii_tensor import interaction_heatmap >>> interactions = scii_tensor.interaction_select_multiple(sct_merge, >>> tme_module=0, sample='p2', >>> cellpair_module=0, >>> lrpair_module=1, n_lr=15, n_cc=15) >>> interaction_heatmap(interactions, figsize=(5, 3), vmax=10) ##visualize selected interactions using circle plot >>> from stereosite.plot.scii_circos import cells_lr_circos, cells_circos, lr_circos >>> cells = adata.obs['cell2loc_anno'].unique() >>> cells_lr_circos(interactions, cells=cells)