Code for the im4MEC model described in the paper 'Interpretable deep learning model to predict the molecular classification of endometrial cancer from haematoxylin and eosin-stained whole-slide images: a combined analysis of the PORTEC randomised trials and clinical cohorts' (The Lancet Digital Health, 2022-12-07).

im4MEC is a deep-learning model for slide-level molecular classification of Endometrial Cancer using the morphological features encoded in H&E whole slide images. It uses self-supervised learning (SSL) to get histopathology domain specific feature representation of tiles, followed by attention-mechanism to identify the tiles with morphological features of high importance towards molecular classification of the Whole Slide Image. im4MEC is interpretable and indentifies morpho-molecular correlates in endometrial cancer.

im4MEC pipeline

Due to issues with OpenSlide dependencies (like this one with pixman) it is recommended to install the project's dependencies using the conda environment.yml rather than individually pip install-ing the dependencies.

The below command creates an unnamed conda environment in your working directory.

conda env create --prefix ./.conda -f environment.yml
conda activate ./.conda

The sample_tiles.py script needs to be executed for every WSI that you want to sample tiles from. In this example, the output tiles images will be stored in moco_tiles/train. Additionally, one quality control (QC) image will be generated for each WSI so the user can evaluate the location of the sampled tiles. This QC image will be stored in moco_tiles/QC in the example.

python sample_tiles.py \
--input_slide /path/to/your/WSI.svs \
--output_dir moco_tiles \
--tile_size 360 \
--n 2048 \
--out_size 224

For example, if you have a list of absolute paths to your WSI files named WSI_FILES.txt and are working on Linux you can use the below command to run sample_tiles.py across all of them. This is a crude, non-parallel approach and we encourage users to create their own wrapper scripts instead to exploit the opportunities for parallelism of their hardware. The sample_tiles.py script does not require a GPU.

cat WSI_FILES.txt | xargs -I WSIFILE echo python sample_tiles.py --input_slide=WSIFILE --output_dir=moco_tiles --tile_size=360 --n=2048 --out_size=224 | bash

We encourage users to read the sample_tiles.py source code for more details of the sampling process and the command line arguments.

This repository contains a fork of the original MoCo codebase, with a few minor modifications:

• Added a cosine learning rate decay routine with warmup.
• Changed the image augmentation pipeline for improved results on HE WSI data.

After you have extracted the tiles that you want to use to train the feature extractor, you can invoke the MoCoV2 training routine using something like the command below. We encourage users to read the MoCo README and source code for more details on how to use it.

python moco/main_moco.py \
moco_tiles \
--lr 0.06 \
--warmup 10 \
--epochs 300 \
--cos \
--batch-size 288 \
--arch resnet50 \
--moco-k 73728 \
--moco-t 0.07 \
--mlp \
--dist-url 'tcp://localhost:10001' \
--multiprocessing-distributed \
--world-size 1 \
--rank 0

Next, each WSI in the dataset needs to be converted into a bag of feature vectors using the feature extractor model that we trained in the previous step using self-supervised learning.

The below example command will use the trained feature extraction model to convert each tile in the WSI into a feature vector. All feature vectors of a WSI will be saved together in a 'feature bag' in the .h5 file format. Additionally, a quality control image will be generated in the same output directory so the user can evaluate the quality of the tissue segmentation and tiling. Both output files will be named after the input WSI, suffixed with *_features.h5 and *_features_QC.png, respectively.

python preprocess.py \
--input_slide /path/to/your/WSI.svs \
--output_dir feature_bags \
--tile_size 360 \
--out_size 224 \
--batch_size 256 \
--checkpoint /path/to/your/feature_extractor.pth.tar \
--backbone resnet50 \
--workers 4

Please refer to the code of the preprocess.py script for an explanation of the command line arguments.

Note that providing a weights checkpoint --checkpoint is optional. If you want to run this step using a feature extractor with ImageNet weights instead, you can use the --imagenet flag.

Note that this is a very compute-intensive script. While it can be run on the CPU, having a GPU available is advisable especially for larger datasets.

Creating feature bags for each WSI in your dataset is an 'embarassingly parallel' problem. Just like with the sample_tiles.py step, we encourage users to create their own wrapper scripts to exploit the potential for parallelism of their systems.

The structure of your experiment should be defined in a CSV file. See labels.csv for an example. This file should contain a single line for each WSI in your dataset.

• The slide_id is an arbitrary identifier for each slide.
• The class column refers to the WSI's class in the classification task
• The label column is the WSI's class but coded as an integer label for use under the hood in Pytorch.
• The fold-0 ... fold-k columns tell train.py whether the WSI should be part of the training or validation set during each fold of the cross validation routine. The number of fold-* columns should correspond to the number of folds you want to use in the k-fold cross validation routine. E.g. when you want k = 6 the columns fold-0 through fold-5 should be present.

To train the attention model that performs the final WSI-level classification, we use the train.py script. This script has a k-fold cross-validation routine built in. The below example command will run the training + validation loop for each set of hyperparameters hardcoded in the main() function of train.py and log the results to Tensorboard. Model checkpoints will not be saved. Note that the cross-validation routine can be parallized if you have the hardware for that by running multiple instances of train.py at a time.

for fold in 0 1 2 3
do
python train.py \
--manifest labels.csv \
--data_dir feature_bags \
--input_feature_size 2048 \
--fold ${fold}
done

Note that the --input_feature_size argument needs to correspond to the size of the feature vectors produced by the feature extractor model. Please refer to the code of the train.py script for an explanation of the other command line arguments.

Once you have established that a certain set of hyperparameters is optimal for your task, you can train the model on the entire training + validation by providing the index of the optimal set of hyperparameters to the --full_training argument. Model checkpoints will be saved in the ./runs directory. The below example command will use the 3rd (index 2) set of hyperparameters.

python train.py \
--manifest labels.csv \
--data_dir feature_bags \
--input_feature_size 2048 \
--full_training 2

Once you have a trained attention model, the attention.py script can be used to generate attention heatmaps for arbitrary WSIs. The below command will create one heatmap for each class in your classification task for the provided WSI.

Note that this script is very resource intensive as it requires running the feature extraction model multiple times across the entire WSI. Using a GPU is advisable.

python attention.py \
--input_slide /path/to/your/WSI.svs \
--output_dir heatmaps \
--manifest labels.csv \
--encoder_checkpoint /path/to/your/feature_extractor.pth.tar \
--encoder_backbone resnet50 \
--attn_checkpoint runs/some_run/5_checkpoint.pt \
--attn_model_size small \
--input_feature_size 2048 \
--tile_size 360 \
--out_size 224 \
--overlap_factor 1 \
--display_level 3