Convolutional neural networks (CNNs) often require very large datasets for robust training and evaluation. As an alternative approach, we introduce deep learning diffusion fingerprinting (DLDF), which treats every voxel as an independent data point, rather than using whole images or patches. We use DLDF to classify diffusion-weighted imaging voxels in a mouse model of glioblastoma, both prior to and in response to Temozolomide chemotherapy. We show that, even with limited training, DLDF can automatically segment brain tumours from normal brain, can distinguish between young and older tumours and that DLDF can detect if a tumour has been treated with chemotherapy.
Glioma mouse model and therapy: 24 mice were injected with GL261 glioma cells in the right striatum. After 13 days of growth, baseline scans were acquired in all mice (day 0). Temozolomide (TMZ) was administered to the treated group of mice (n = 12). Vehicle was administered to the control group (n = 12). After nine more days (day 9), follow-up images were acquired.
MRI: A PGSE diffusion-rich dataset with 46 diffusion weightings and a 42-direction DTI was acquired on a 9.4T Agilent scanner [2]. Five coronal slices were acquired in each animal. Slice-matched, T1-weighted post-Gd-DTPA images were acquired (Figure 1a) after diffusion MRI.
Deep Learning: For training the neural network, four categories were manually identified on Gd-DTPA images: 1) outside of the brain, 2) normal-appearing brain, 3) untreated tumour at day 0 and day 9, 4) treated tumour at day 9 (Figure 1b). Diffusion fingerprints were produced for each pixel consisting of 232 ordered data points (Figure 2). Python libraries (Keras and TensorFlow [3]) were used to construct a neural network with 5 hidden layers. Each hidden layer was regularized with 20% dropout, and used ReLU activation and normally-distributed initialisation. 50% of the data (6 control, 6 treated) were randomly assigned to a training set, which were further refined to include equal numbers of fingerprints from each category (total number of fingerprints = 6,608). Neural network training was performed using stochastic gradient-descent optimisation with a mean squared error loss function. Training was performed for 50,000 epochs, with 10,000 steps in each. 5 animals per group were used for evaluation.
For training, the T1-weighted Gd-DTPA images (Figure 1a) were used to create category masks (Figure 1b) which were applied to the multi-dimensional DWI images (Figure 2a) to generate pixel-wise diffusion fingerprints (Figure 2c). Upon visual inspection of the diffusion fingerprints, some subtle differences could be observed between fingerprints from normal brain (green) and tumour (yellow/red).
The neural network training accuracy was 98.65%. The evaluation accuracy was 98.54%. A strong agreement was observed between manually segmented category masks (Figure 1b) and the evaluation DLDF maps (Figure 1c), demonstrating the ability of the neural network to automatically segment the tumours. The neural network was also able to distinguish between tumours based on age (Figure 1e) – voxels within day 0/day 9 tumours were predominantly classified as day 0/day 9 tumours, respectively. In comparison, ADC alone was unable to stratify all pre-therapy tumours from day 9 tumours (Figure 3).
DLDF category maps at day 9 showed a strong spatial accordance with histological slices (Figure 4). For example, treated tumours generally showed a distinctive rim of ‘untreated’ pixels (yellow) at the periphery of the tumour, which could correspond to regions in H&E images where tumour transitions into the normal brain (Figure 4d). In some untreated brains, the DLDF category maps showed tumour invasion into the contralateral hemisphere along the corpus callosum, which also showed enhanced GFAP-staining on histology (Figure 5).
[1] Litjens G, Kooi T, Bejnordi BE, et al. A survey on deep learning in medical image analysis. arXiv preprint arXiv:170205747 (2017)
[2] Panagiotaki E, Walker-Samuel S, Siow B, et al. Noninvasive quantification of solid tumor microstructure using VERDICT MRI. Cancer Research (2014).
[3] Abadi M, Agarwal A, Barham P, et al. Tensorflow: Large-scale machine learning on heterogeneous distributed systems. arXiv preprint arXiv:160304467 (2016)