Abstract
The quantity of leukocytes in papillary thyroid carcinoma (PTC) potentially have prognostic and treatment predictive value. Here, we propose a novel method for training a convolutional neural network (CNN) algorithm for segmenting leukocytes in PTCs. Tissue samples from two retrospective PTC cohort were obtained and representative tissue slides from twelve patients were stained with hematoxylin and eosin (HE) and digitized. Then, the HE slides were destained and restained immunohistochemically (IHC) with antibodies to the pan-leukocyte anti CD45 antigen and scanned again. The two stain-pairs of all representative tissue slides were registered, and image tiles of regions of interests were exported. The image tiles were processed and the 3,3'-diaminobenzidine (DAB) stained areas representing anti CD45 expression were turned into binary masks. These binary masks were applied as annotations on the HE image tiles and used in the training of a CNN algorithm. Ten whole slide images (WSIs) were used for training using a five-fold cross-validation and the remaining two slides were used as an independent test set for the trained model. For visual evaluation, the algorithm was run on all twelve WSIs, and in total 238,144 tiles sized 500 × 500 pixels were analyzed. The trained CNN algorithm had an intersection over union of 0.82 for detection of leukocytes in the HE image tiles when comparing the prediction masks to the ground truth anti CD45 mask. We conclude that this method for generating antibody supervised annotations using the destain-restain IHC guided annotations resulted in high accuracy segmentations of leukocytes in HE tissue images.
Highlights
P APILLARY thyroid carcinoma (PTC), the most common variant of thyroid cancer, shows an increase in incidence and is about three times more common in women [1]–[3]
The trained convolutional neural network (CNN) algorithm was tested on 351 image tiles sized 500 × 500 pixels
For each of these tiles, the probability masks generated by the CNN algorithm were turned into binary masks and compared to the ground truth mask based on the anti CD45 DAB staining
Summary
P APILLARY thyroid carcinoma (PTC), the most common variant of thyroid cancer, shows an increase in incidence and is about three times more common in women [1]–[3]. The immune response plays a crucial role in the defense against the development of cancer. There is evidence that inflammatory cells can be actively tumor promoting [5]. The inflammatory milieu of PTC plays a crucial role in tumor progression, metastasis and recurrence of thyroid cancer [6], [7]. The presence of immune cells has been shown to correlate with a favorable outcome of PTC [8], [9]. The prognostic significance of specific immune cells in PTC has been studied by analyzing immunological parameters specific to certain cells [10]. Several specific immunological markers have been shown to be prognostically significant, including CD8 and PD-L1 [11]
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