finest 0.0.7

FineST: Fine-grained Spatial Transcriptomic

A statistical model and toolbox to identify the super-resolved ligand-receptor interaction with spatial co-expression (i.e., spatial association). Uniquely, FineST can distinguish co-expressed ligand-receptor pairs (LR pairs) from spatially separating pairs at sub-spot level or single-cell level, and identify the super-resolved ligand-receptor interaction (LRI).

It comprises three components (Training-Imputation-Discovery) after HE image feature is extracted:

• Step0: HE image feature extraction

• Step1: Training FineST on the within spots

• Step2: Super-resolution spatial RNA-seq imputation

• Step3: Fine-grained LR pair and CCC pattern discovery

Installation

FineST is available through PyPI. To install, type the following command line and add -U for updates:

pip install -U FineST

Alternatively, install from this GitHub repository for latest (often development) version (time: < 1 min):

pip install -U git+https://github.com/StatBiomed/FineST

Installation using Conda

$ git clone https://github.com/StatBiomed/FineST.git
$ conda create --name FineST python=3.8
$ conda activate FineST
$ cd FineST
$ pip install -r requirements.txt

Typically installation is completed within a few minutes. Then install pytorch, refer to pytorch installation.

$ conda install pytorch=1.7.1 torchvision torchaudio cudatoolkit=11.0 -c pytorch

Verify the installation using the following command:

python
>>> import torch
>>> print(torch.__version__)
>>> print(torch.cuda.is_available())

Get Started for Visium or Visium HD data

Usage illustrations:

• For Visium, using a single slice of 10x Visium human nasopharyngeal carcinoma (NPC) data.

• For Visium HD, using a single slice of 10x Visium HD human colorectal cancer (CRC) data with 16-um bin.

Step0: HE image feature extraction (for Visium)

Visium (v2) measures about 5k spots across the entire tissue area. The diameter of each individual spot is roughly 55 micrometers (um), while the center-to-center distance between two adjacent spots is about 100 um. In order to capture the gene expression profile across the whole tissue ASSP,

Firstly, interpolate between spots in horizontal and vertical directions, using Spot_interpolate.py.

python ./FineST/Spot_interpolate.py \
   --data_path ./Dataset/NPC/ \
   --position_list tissue_positions_list.csv \
   --dataset patient1

with Input: tissue_positions_list.csv - Locations of within spots (n), and Output: _position_add_tissue.csv- Locations of between spots (m ~= 3n).

Then extracte the within spots HE image feature embeddings using HIPT_image_feature_extract.py.

python ./FineST/HIPT_image_feature_extract.py \
   --dataset AH_Patient1 \
   --position ./Dataset/NPC/patient1/tissue_positions_list.csv \
   --image ./Dataset/NPC/patient1/20210809-C-AH4199551.tif \
   --output_path_img ./Dataset/NPC/HIPT/AH_Patient1_pth_64_16_image \
   --output_path_pth ./Dataset/NPC/HIPT/AH_Patient1_pth_64_16 \
   --patch_size 64 \
   --logging_folder ./Logging/HIPT_AH_Patient1/

Similarlly, extracte the between spots HE image feature embeddings using HIPT_image_feature_extract.py.

python ./FineST/HIPT_image_feature_extract.py \
   --dataset AH_Patient1 \
   --position ./Dataset/NPC/patient1/patient1_position_add_tissue.csv \
   --image ./Dataset/NPC/patient1/20210809-C-AH4199551.tif \
   --output_path_img ./Dataset/NPC/HIPT/NEW_AH_Patient1_pth_64_16_image \
   --output_path_pth ./Dataset/NPC/HIPT/NEW_AH_Patient1_pth_64_16 \
   --patch_size 64 \
   --logging_folder ./Logging/HIPT_AH_Patient1/

HIPT_image_feature_extract.py also output the execution time:

• The image segment execution time for the loop is: 8.153 seconds

• The image feature extract time for the loop is: 35.499 seconds

Input files:

• 20210809-C-AH4199551.tif: Raw histology image

• patient1_position_add_tissue.csv: “Between spot” (Interpolated spots) locations

Output files:

• NEW_AH_Patient1_pth_64_16_image: Segmeted “Between spot” histology image patches (.png)

• NEW_AH_Patient1_pth_64_16: Extracted “Between spot” image feature embeddiings for each patche (.pth)

Step0: HE image feature extraction (for Visium HD)

Visium HD captures continuous squares without gaps, it measures the whole tissue area.

python ./FineST/HIPT_image_feature_extract.py \
   --dataset HD_CRC_16um \
   --position ./Dataset/CRC/square_016um/tissue_positions.parquet \
   --image ./Dataset/CRC/square_016um/Visium_HD_Human_Colon_Cancer_tissue_image.btf \
   --output_path_img ./Dataset/CRC/HIPT/HD_CRC_16um_pth_32_16_image \
   --output_path_pth ./Dataset/CRC/HIPT/HD_CRC_16um_pth_32_16 \
   --patch_size 32 \
   --logging_folder ./Logging/HIPT_HD_CRC_16um/

HIPT_image_feature_extract.py also output the execution time:

• The image segment execution time for the loop is: 62.491 seconds

• The image feature extract time for the loop is: 1717.818 seconds

Input files:

• Visium_HD_Human_Colon_Cancer_tissue_image.btf: Raw histology image (.btf Visium HD or .tif Visium)

• tissue_positions.parquet: Spot/bin locations (.parquet Visium HD or .csv Visium)

Output files:

• HD_CRC_16um_pth_32_16_image: Segmeted histology image patches (.png)

• HD_CRC_16um_pth_32_16: Extracted image feature embeddiings for each patche (.pth)

Step1: Training FineST on the within spots

Step2: Super-resolution spatial RNA-seq imputation

Step3: Fine-grained LR pair and CCC pattern discovery

Detailed Manual

The full manual is at finest-rtd-tutorial for installation, tutorials and examples.

• Interpolate between-spots among within-spots by FineST (For Visium dataset).

• Crop region of interest (ROI) from HE image by FineST (Visium or Visium HD).

• Sub-spot level (16x resolution) prediction by FineST (For Visium dataset).

• Sub-bin level (from 16um to 8um) prediction by FineST (For Visium HD dataset).

• Super-resolved ligand-receptor interavtion discovery by FineST.