Purpose: Recent studies suggest an inverse association between cancer in general and Alzheimer’s disease (AD), suggesting that there are common factors in these diseases. In this study, we demonstrate a complete workflow to streamline the analysis of stained histological brain samples to reduce user-dependence. As a proof of concept, we investigate the influence of Amyloid Beta (Aβ) plaques and hyperphosphorylated Tau protein (pTau), hallmarks of AD, in Glioblastoma (GBM) patients. Methods: The automated digital histology processing workflow is demonstrated on a 10-patient cohort. First, tissue samples were taken at autopsy from regions known to be common Aβ and pTau hotspots. The tissue samples were then subclassified regionally using a ResNet50 neural network to separate gray and white matter, and tissue processing artifacts. Positively stained areas were then automatically detected and analyzed between subjects. We evaluated the automatic sample classification using a 5-fold cross validation. Results: Cross validation achieved an overall accuracy of 93.88%. Positively stained sample regions were automatically detected and present in six of ten patients for Aβ and in three of ten patients for pTau. General accumulation of both pTau and Aβ calculated over all samples correlated with the age of the patient, and showed decreased accumulation in the brain hemisphere where the primary tumor was located. Conclusion: The proposed method for processing histological samples of the brain automates the time consuming and error prone manual segmentation of grey matter and removal of artifacts. Our study highlights hemispheric differences in pTau and Aβ accumulation. Future studies of pTau and Aβ in the presence of GBM will help to understand how tumor location and growth affect micro-environmental factors in larger cohorts of patients.
Purpose: Multiplex staining allows molecular co-localization analysis of same cell or tissue sections and maximizes the amount of data acquired from individual samples. We developed a non-linear registration framework for automated alignment of whole mount multiplexed histology images of prostate cancer. To achieve a precise automatic fit of bleached and restained high resolution tissue samples, a patch-based approach is proposed to improve annotation speed and analysis. Methods: The three-step co-registration process begins with a coarse low-resolution registration of the IHC stained image to the fixed H&E-stained image. The initial registration is then refined separately for each high-resolution patch using a smaller search window. Finally, registered patches are stitched back together using speeded up robust features (SURF). We apply the method to five multiplex whole mount prostate histology slides. To determine its effectiveness, we compare the automatic registration to the initial coarse registration and a manual control point based-method varying the number of control points. Results: For the control point-based approach, 25, 50, and 100 manually placed set landmarks resulted in a decrease of - 76.20%, -75.9% and -75.48% of the root mean squared error (RMSE), respectively. Compared to the initial registration an improvement of -76.29% of RMSE can be seen, illustrating the potential benefits of a patch-based automatic approach. Conclusion: The proposed method achieved excellent registration of the IHC to the input image. The automated method for registration of multiplexed histology images achieves high accuracy in shorter time and with greater reproducibility than conventional registration approaches and semi-automatic control points without the need for time consuming and subjective manual control-point setting. The accuracy of the fit especially improves in complex areas close to tissue tears and folds.
Purpose: Prostate cancer primarily arises from the glandular epithelium. Histomophometric techniques have been used to assess the glandular epithelium in automated detection and classification pipelines; however, they are often rigid in their implementation, and their performance suffers on large datasets where variation in staining, imaging, and preparation is difficult to control. The purpose of this study is to quantify performance of a pixelwise segmentation algorithm that was trained using different combinations of weak and strong stroma, epithelium, and lumen labels in a prostate histology dataset.Approach: We have combined weakly labeled datasets generated using simple morphometric techniques and high-quality labeled datasets from human observers in prostate biopsy cores to train a convolutional neural network for use in whole mount prostate labeling pipelines. With trained networks, we characterize pixelwise segmentation of stromal, epithelium, and lumen (SEL) regions on both biopsy core and whole-mount H&E-stained tissue.Results: We provide evidence that by simply training a deep learning algorithm on weakly labeled data generated from rigid morphometric methods, we can improve the robustness of classification over the morphometric methods used to train the classifier.Conclusions: We show that not only does our approach of combining weak and strong labels for training the CNN improve qualitative SEL labeling within tissue but also the deep learning generated labels are superior for cancer classification in a higher-order algorithm over the morphometrically derived labels it was trained on.
Sean McGarry, John Bukowy, Kenneth Iczkowski, Allison Lowman, Michael Brehler, Samuel Bobholz, Andrew Nencka, Alex Barrington, Kenneth Jacobsohn, Jackson Unteriner, Petar Duvnjak, Michael Griffin, Mark Hohenwalter, Tucker Keuter, Wei Huang, Tatjana Antic, Gladell Paner, Watchareepohn Palangmonthip, Anjishnu Banerjee, Peter LaViolette
Purpose: Our study predictively maps epithelium density in magnetic resonance imaging (MRI) space while varying the ground truth labels provided by five pathologists to quantify the downstream effects of interobserver variability.
Approach: Clinical imaging and postsurgical tissue from 48 recruited prospective patients were used in our study. Tissue was sliced to match the MRI orientation and whole-mount slides were stained and digitized. Data from 28 patients (n = 33 slides) were sent to five pathologists to be annotated. Slides from the remaining 20 patients (n = 123 slides) were annotated by one of the five pathologists. Interpathologist variability was measured using Krippendorff’s alpha. Pathologist-specific radiopathomic mapping models were trained using a partial least-squares regression using MRI values to predict epithelium density, a known marker for disease severity. An analysis of variance characterized intermodel means difference in epithelium density. A consensus model was created and evaluated using a receiver operator characteristic classifying high grade versus low grade and benign, and was statistically compared to apparent diffusion coefficient (ADC).
Results: Interobserver variability ranged from low to acceptable agreement (0.31 to 0.69). There was a statistically significant difference in mean predicted epithelium density values (p < 0.001) between the five models. The consensus model outperformed ADC (areas under the curve = 0.80 and 0.71, respectively, p < 0.05).
Conclusion: We demonstrate that radiopathomic maps of epithelium density are sensitive to the pathologist annotating the dataset; however, it is unclear if these differences are clinically significant. The consensus model produced the best maps, matched the performance of the best individual model, and outperformed ADC.
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