Pathological estimation of tumor necrosis after chemotherapy is essential for patients with osteosarcoma. This study reports the first fully automated tool to assess viable and necrotic tumor in osteosarcoma, employing advances in histopathology digitization and automated learning. We selected 40 digitized whole slide images representing the heterogeneity of osteosarcoma and chemotherapy response. With the goal of labeling the diverse regions of the digitized tissue into viable tumor, necrotic tumor, and non-tumor, we trained 13 machine-learning models and selected the top performing one (a Support Vector Machine) based on reported accuracy. We also developed a deep-learning architecture and trained it on the same data set. We computed the receiver-operator characteristic for discrimination of non-tumor from tumor followed by conditional discrimination of necrotic from viable tumor and found our models performing exceptionally well. We then used the trained models to identify regions of interest on image-tiles generated from test whole slide images. The classification output is visualized as a tumor-prediction map, displaying the extent of viable and necrotic tumor in the slide image. Thus, we lay the foundation for a complete tumor assessment pipeline from original histology images to tumor-prediction map generation. The proposed pipeline can also be adopted for other types of tumor.
Pathologists often deal with high complexity and sometimes disagreement over osteosarcoma tumor classification due to cellular heterogeneity in the dataset. Segmentation and classification of histology tissue in H&E stained tumor image datasets is a challenging task because of intra-class variations, inter-class similarity, crowded context, and noisy data. In recent years, deep learning approaches have led to encouraging results in breast cancer and prostate cancer analysis. In this article, we propose convolutional neural network (CNN) as a tool to improve efficiency and accuracy of osteosarcoma tumor classification into tumor classes (viable tumor, necrosis) versus nontumor. The proposed CNN architecture contains eight learned layers: three sets of stacked two convolutional layers interspersed with max pooling layers for feature extraction and two fully connected layers with data augmentation strategies to boost performance. The use of a neural network results in higher accuracy of average 92% for the classification. We compare the proposed architecture with three existing and proven CNN architectures for image classification: AlexNet, LeNet, and VGGNet. We also provide a pipeline to calculate percentage necrosis in a given whole slide image. We conclude that the use of neural networks can assure both high accuracy and efficiency in osteosarcoma classification.
Osteosarcoma is one of the most common types of bone cancer in children. To gauge the extent of cancer treatment response in the patient after surgical resection, the H&E stained image slides are manually evaluated by pathologists to estimate the percentage of necrosis, a time consuming process prone to observer bias and inaccuracy. Digital image analysis is a potential method to automate this process, thus saving time and providing a more accurate evaluation. The slides are scanned in Aperio Scanscope, converted to digital Whole Slide Images (WSIs) and stored in SVS format. These are high resolution images, of the order of 109 pixels, allowing up to 40X magnification factor. This paper proposes an image segmentation and analysis technique for segmenting tumor and non-tumor regions in histopathological WSIs of osteosarcoma datasets. Our approach is a combination of pixel-based and object-based methods which utilize tumor properties such as nuclei cluster, density, and circularity to classify tumor regions as viable and non-viable. A K-Means clustering technique is used for tumor isolation using color normalization, followed by multi-threshold Otsu segmentation technique to further classify tumor region as viable and non-viable. Then a Flood-fill algorithm is applied to cluster similar pixels into cellular objects and compute cluster data for further analysis of regions under study. To the best of our knowledge this is the first comprehensive solution that is able to produce such a classification for Osteosarcoma cancer. The results are very conclusive in identifying viable and non-viable tumor regions. In our experiments, the accuracy of the discussed approach is 100% in viable tumor and coagulative necrosis identification while it is around 90% for fibrosis and acellular/hypocellular tumor osteoid, for all the sampled datasets used. We expect the developed software to lead to a significant increase in accuracy and decrease in inter-observer variability in assessment of necrosis by the pathologists and a reduction in the time spent by the pathologists in such assessments.
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