AxonFinder: Automated segmentation of tumor innervating neuronal fibers

Neurosignaling is increasingly recognized as a critical factor in cancer progression, where neuronal innervation of primary tumors contributes to the disease's advancement. This study focuses on segmenting individual axons within the prostate tumor microenvironment, which have been challenging to detect and analyze due to their irregular morphologies. We present a novel deep learning-based approach for the automated segmentation of axons, AxonFinder, leveraging a U-Net model with a ResNet-101 encoder, based on a multiplexed imaging approach. Utilizing a dataset of whole-slide images from low-, intermediate-, and high-risk prostate cancer patients, we manually annotated axons to train our model, achieving significant accuracy in detecting axonal structures that were previously hard to segment. Our method achieves high performance, with a validation F1-score of 94 % and IoU of 90.78 %. Besides, the morphometric analysis that shows strong alignment between manual annotations and automated segmentation with nerve length and tortuosity closely matching manual measurements. Furthermore, our analysis includes a comprehensive assessment of axon density and morphological features across different CAPRA-S prostate cancer risk categories revealing a significant decline in axon density correlating with higher CAPRA-S prostate cancer risk scores. Our paper suggests the potential utility of neuronal markers in the prognostic assessment of prostate cancer in aiding the pathologist's assessment of tumor sections and advancing our understanding of neurosignaling in the tumor microenvironment.